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Communication, facilitation, ​powerspeaking blog: tips and strategies for crafting presentations, how to present to senior executives [23 proven strategies].

by Carrie Beckstrom     Jun 4, 2021 10:45:46 AM

If you’re preparing to make a pitch to senior executives, you can throw most of what you’ve learned about making presentations in the trash.

Why? Because this is an entirely different audience. An executive presentation is not about inspiring and persuading, but is instead about presenting meaningful data to top decision-makers as quickly and concisely as possible. As a matter of fact, you should start with your bottom line (what you need from them). And work your way from there.

As a company that lives and breathes communication, we are dedicated to transforming your organization's communication skills, so you can watch people succeed at engaging and influencing others, achieving results, and driving business to new heights.

And today, we are going to reveal our favorite tips for a powerful executive presentation to decision-making leaders to help you get your point across efficiently, earn your audience’s attention, and know when and how to go in for the "ask."

So if you want more success from your business presentations, then you'll love this list of tips, techniques, and strategies.

Keep reading to get started with our tried and true tips!

Don't have time to read the whole checklist right now? 

Save the PDF version to your desktop and read when it's convenient for you.

Tip 1 Understand the executive mindset

Tip 2 Consider executives’ personality styles

Tip 3 Give yourself enough time to gather data

Tip 4 Find and leverage a sponsor

Tip 5 Prepare for possible questions or objections

Tip 6 Follow the 10/30 Rule

Tip 7 Set the context and confirm the time slot

Tip 8 Use the PREP model if your time gets cut

Tip 9 State the outcome you want early

Tip 10 Always use a valid business reason

Tip 11 Follow with the essential data—and do a deep dive only if asked

Tip 12 Focus on analytical proof

Tip 13 Be proactive and transparent when delivering unpleasant news

Tip 14 Remember that executives want a dialogue—not a data dump

Tip 15 Read the room

Tip 16 Use the Headline + Evidence model

Tip 17 Paraphrase complex questions to gain clarity

Tip 18 Address heated feedback and challenging questions in the moment

Tip 19 Answer the question-behind-the-question (before it is even asked)

Tip 20 Rely on the “notice and ask” strategy to refocus the discussion

Tip 21 Be personal to re-engage a disengaged executive

Tip 22 Acknowledge disagreements between top executives, but don't intervene

Tip 23 Conclude with a quick summary and resolve any noteworthy issues before your final ask

Let's dive right in.

Before you try to deliver your next presentation to executives, keep this in mind...

Most people fail with their first attempt to wow the top executives for one simple reason:

C-level executives demand an entirely different approach from presenters than does a general business audience. SVP of Strategy and Business Development at Workato , Markus Zirn says,  

“You have to throw out everything you learned from traditional presentation training. It’s not about making a speech that educates, persuades, inspires, or leads. It’s about raw decision-making.”

With that little caveat out of the way, it’s time for our first strategy…

Understand the Executive Mindset

Executives are impatient, bottom-line-focused leaders who don't care about the minutiae of your project or expertise. They want you to get to the point, step away from your slide deck, and be ready for a dynamic dialogue.

During your presentation, they might multitask and switch topics quickly, make calculations in their heads at lightning speed, and they may be challenging when analyzing data. Be ready for this.  

Understanding the executive's mindset as you are going in for the pitch will help you succeed. Watch the video below on Audience Analysis and getting started.

Here's just one executive you'll hear from...

"If it is a C-suite presentation and you’re actually asking for an outcome, you need to know what are the hot points of the CFO, or the treasurer’s office, or the VP of finance."

Consider executives’ personality styles

Think of each executive who will be in the room.

How do they prefer to have information presented to them?  Where are they on the risk profile?  Are they highly conservative, or are they innovative and open to experimentation?  How will your proposal affect their areas of responsibility? 

Starting with your audience in mind helps you strategize how to present your information.

Give yourself enough time to gather data

Weeks ahead of your presentation to executives, you may need to: meet with people in various functional areas for input and/or buy-in; check in with your manager; research executives’ hot buttons; and prepare slides or documents to send ahead. Don’t underestimate the time it will take to do all of this.

And speaking of being prepared, make sure you’ve considered the environment in which you’ll likely be with them—namely, virtual. If presenting to executives weren’t challenging enough, today’s digital interactions present another layer of things to consider.

Check out this great article, “ Digital Communication Skills: Is Your Message Getting Across? ,” by digital presenting and training guru Cindy Huggett. Great suggestions not only for putting your best digital self forward with senior executives, but also tips you can use in any virtual-meeting situation.

Find and leverage a sponsor

Find someone who knows the group you’re presenting to who is willing to not only help you prepare as your guide, but can also advocate for you during the meeting. Use your sponsor to check the validity of your logic ahead of time, rehearse with and get feedback from, and agree on when you should look to them for help during the executive meeting. 

Debrief with your sponsor after the meeting to gain insights and lessons you can use in the future.

Prepare for possible questions or objections

Consider including a slide that states the answers to anticipated questions or concerns rather than waiting for them to be brought up. This is a great way to demonstrate that you’ve kept your audience in mind when preparing your presentation.

Follow the 10/30 Rule

If you have 30 minutes on the executives’ meeting agenda, plan to present 10 minutes of material. The remainder of the time will likely be devoted to discussion and interaction and not necessarily in sequence.

R emember that your role is to facilitate a conversation, not just deliver a presentation.

Set the context and confirm the time slot

You don’t need an entertaining, attention-getting opening, but you do need to set the context as you start and get an agreement on timing. Thirty minutes originally may have been planned, but the meeting timeline can change. 

Start by saying, “I understand we have scheduled 30 minutes. Does that timing still work?”

Rajeev Goel, Co-Founder and CEO at PubMatic, and Monica Poindexter, Vice President, Inclusion & Diversity/Employee Relations at Lyft, will give you tips on overcoming the presentation challenges in regards to context.

Use the PREP model if your time gets cut

Don’t be surprised if what you thought would be a 30-minute meeting is cut to 10 minutes because the executives have another more urgent matter to discuss. Use the “PREP” model to craft your abbreviated pitch before the meeting:

State the outcome you want early

State the outcome you want from your presentation early. What do you want the executives to say “yes” to?

Be specific. For example, “I want your support” is vague. “I want you to approve a $500,000 investment in new centrifuge equipment this quarter” is specific.

Get that key message out first...

Always use a valid business reason

Clearly state what value or ROI your ask will bring in terms of revenue increase, cost reduction, protection from risk, competitive advantage, or other business drivers. It can’t be just a “nice to have” proposal. 

For example...

“The bottom line is, we’re seeking your approval for five new FTEs in account management roles for an investment of $750,000. The reason is that we have a $2M revenue opportunity with the launch of [XYZ project]. These account managers will focus on the top 20% of our customer base to ensure we maximize the sales opportunity.”

Follow with the essential data—and do a deep dive only if asked

Instead of drowning listeners in data, lead with the top two to three key points and supporting evidence that best support your ask and business reason. The key points should be answers to the primary questions you think the executives will have about your proposal. 

Keep in mind that execs want the 10,000-foot view and how it affects the organization's bottom line.  

Focus on analytical proof

Senior decision-makers don’t need to be entertained. So while your logic may follow a “storyline,” hard data is what will be more convincing.

If you have a customer success story or a brief anecdote that supports your point, use it. But remember, unlike a general audience, the C-level executives in the room will grow impatient with stories unless they’re succinct and tie directly to your proposal.

Be proactive and transparent when delivering unpleasant news

There are four steps to presenting bad news:

• Get it out at the beginning
• Be candid about what happened
• Explain how you will fix it
• Offer options and recommendations. 

You can use this 4-step framework to present bad news in a way that speaks to executives.

Let's take a look at a hypothetical example. You have to explain to them why a consultant they hired quit, and you need to rectify the situation...

1. G et it out at the beginning

The consultant you approved quit unexpectedly.  A response to the audit she was supposed to help us with is due next week. Today I want to talk about the path forward. 

2. Be candid and explain how you will fix the situation

By the end of this meeting, I’d like your approval to:

Respond to the audit in a generic way with help from the audit team

Collaborate with the internal process improvement team to assess our local processes 

Collaborate with my counterpart to create one agreement that combines the best aspects of our process and theirs 

3. Offer options on why it makes business sense 

No additional cost of hiring a consultant

We’ll be able to leverage the existing processes to create a process that is consistent across the globe

4. Offer final recommendations   

With all of that in mind, today I’d like to talk about how we should change our audit process and why I’m confident this plan will receive regulatory approval.

Is there anything else you’d like me to add?

If you have bad news, don’t try to gloss over it. Use the framework to present it in a way that speaks to executives.

Remember that executives want a dialogue—not a data dump

Expect questions and energetic discussions to occur before you have presented all of your material.

(Remember the 10/30 Rule: If you have 30 minutes, prepare only 10 minutes’ worth of “presentation.”) Respond to questions when they come up.

Use facilitation skills to refocus the discussion after a topic change.

Here are a few more tips to enhance your facilitation skills:

Read the room

Notice the process of the discussion as well as the content. When there is cross-talk, interruptions, or rapid-fire questions, you can observe first and then choose an appropriate intervention.

Reading the room can be especially challenging in virtual meetings. You want to encourage everyone to turn on their webcam. Solicit feedback via chat. Stop the presentation and do a quick “reality check” if you notice disengagement or tension.

Use the Headline + Evidence model

Again, this is a time-pressed audience that wants brief, focused answers to their questions. Use the “Headline Response” model to help you do just that.

Paraphrase complex questions to gain clarity

Instead of restating an executive’s question verbatim, use synonyms when you paraphrase.

Remember, you don’t know what is on someone’s mind. So, always check for accuracy after paraphrasing. “Did I get that right?” or just “Right?” gives the questioner a chance to agree or disagree. If your understanding wasn’t accurate, try again.

Address heated feedback and challenging questions in the moment

Especially if someone is upset, a deeper level of listening is required. Reflect on both the content and the feeling you noticed (skeptical, concerned, frustrated, etc.) and answer the question as best you can.

Always check to see if you got it right. Never say, “I know how you feel,” because you don’t, actually. 

Answer the question-behind-the-question (before it is even asked)

If you get resistance or objections to your ideas, often executives are concerned with key business drivers such as top-line and bottom-line impact, competitive positioning, and risk management.

When you paraphrase their questions, check for these concerns and be ready to address them directly.

Listen for the question-behind-the-question.

Rely on the “notice and ask” strategy to refocus the discussion

You are not in a position to tell the executives what to do.

But if the discussion goes in a different direction, you can say, “I notice our discussion is now focused on X. May I get your guidance? Would you like to keep going in this direction, or would you like to go back to our topic?” 

Be personal to re-engage a disengaged executive

You don’t want to catch anyone off guard, so give a little context when you ask someone directly for their thoughts (as a re-engagement strategy).

For example, “Jaime, given that what we are trying to do here is to reduce response time, in your experience, will this new approach work in the field?” 

Acknowledge disagreements between top executives, but don't intervene

You may look to your sponsor to help calm things down, but don’t intervene.

Conclude with a quick summary and resolve any noteworthy issues before your final ask

As you end your presentation, summarize and check to be sure you are clear on what decisions or agreements have been made and what your next steps will be. 

“I explained x and y; you also brought up a and b. Do I have your go-ahead now to purchase the new equipment for $500,000 this quarter?”

Presenting to senior executives is often a nerve-wracking, high-stakes situation. But as these leaders will tell you, it can be a great opportunity to grow, and to increase your value to the organization.

I hope you’ve found these tips enlightening and helpful. L earning to think like senior executives and present to them means learning new habits—and breaking old ones. But wow, I know it’s hard. And we present a lot of information to take in with our 23 Tips.

So, here’s one final piece of advice: start small. Pick one or two tips to apply to your next presentation. And check out this quick-read article, “ 4 Steps to Making New Habits Stick ,” by Cindy Solomon, founder, and CEO of the Courageous Leadership Institute. It will help you learn how to make the most of small steps toward new habits.

But before you get started using them for your next presentation, I’d like to hear from you.

Which presentation tip from this list was your favorite?

...Or maybe you have a suggestion that I didn't cover here?

Either way, let me know by leaving a comment below.

Join Us in Conversation . . .

Best-Practice Crisis Communication

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10 Tips For Impressive Presentations To Senior Leadership And Executives

• Posted on February 7, 2022

10 PRESENTATION TIPS TO IMPRESS SENIOR LEADERS

Tip 1: prepare, tip 2: begin with the end in mind.

• What do you want this audience to feel when they’re done?
• What do you want them to know when you’re done?
• What do you want them to do, when you’re done?

TIP 3: BE CLEAR AND CONCISE

Tip 4: tell stories, tip 5: keep slides minimal.

Provide extra information in your slide notes

TIP 6: CHANNEL NERVOUS ENERGY INTO USABLE ENERGY

The best way to feel confident is to practice.

5 to 7 full rehearsals

TIP 7: PRE-PRESENTATION WARM UP

Breathe low and slow.

WARM UP YOUR BODY

Warm up your mouth, tip 8: have vocal executive presence, tip 9: commanding the space.

Presentation power posing

TIP 10: AUTHENTIC SPEAKER PERSONA

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How to Create a Compelling C-Level Presentation

“Your team understood the needs (of our client) and produced collaterals that were professional, balanced, and delivered the key messages without going overboard.”

Although this was a few years ago, I always remember and cherish these words of praise from one of our C-level (titles typically beginning with ‘C’ - Chief Executive Officer, Chief Operation Officer etc.) clients of the management consulting firm I was working for. Coming from him, it certainly meant a lot because I knew the effort it took to design a presentation for them.

When I first began designing presentations, I found myself learning the ropes quite quickly. The primary concepts behind creating impactful presentations – telling a story, crisp messaging, visually appealing design – flowed naturally to me.

But I quickly realized that working on senior executive level presentations was a different ball game altogether. The clients always had processes in place, clear objectives, and tight timelines. The biggest challenge was to connect with the target executives through the presentation within an extremely short time.

The rules for crafting a C-level presentation are very different and require a considerable amount of thought, strategy, and effort. It is always about quality and not volume – the right information over maximum data.

Working on my first senior leadership presentation was utterly chaotic and nerve-racking. As they would be. It got better after that, and when I received this feedback, I knew I had progressed.

After working on countless executive presentations and interacting with numerous top-level leaders, I have arrived at my own roadmap of designing them effectively and efficiently.

The four cornerstones of a successful presentation hinge on:

• making the conclusion your introduction
• anticipating every single question
• being ready with short and long versions
• creating an impactful summary.

I just wish I had known these earlier and avoided much grief!

Make the Conclusion Your Introduction

“We found that, indeed, time is the scarcest resource.” Michael Porter, a Harvard professor, tells CNBC describing a study he spearheaded on “How CEOs Manage Time."

Porter’s study found that 75% of a CEO’s time was taken up well in advance illustrating just how little time executives have when it comes to meetings and presentations. The learning? Make presentations slim enough to fit into their narrow time slots.

Skip the storytelling and bump up the concluding points to your introduction to establish the point of the meeting right away. It’s imperative to get straight to the point when presenting to time-crunched executives by telling them the whys and the hows upfront with supporting information. That means you would need to do an enormous amount of work filtering the information down to just the right amount.

“In our debriefs, CEOs confessed that one-hour meetings could often be cut to 30 or even 15 minutes,” says Porter. Your presentation should be able to convey everything the CEO needs to know within the first five minutes as there are bound to be questions.

Pro tip – Begin working backwards on the presentation in an inverted pyramid fashion. Start with the goal, the end results, and find the data to support it.

Anticipate Questions and Interruptions

Senior-level executives are an impatient lot. They process information in seconds and form mental maps of a situation as soon as five minutes into the presentation. They are always ahead of the presenter, and that’s bound to bring up questions and interruptions.

It is extremely important to ask multiple questions from an executive’s point of view while designing the presentation.

“What will she not like in this slide?” “Is there ambiguity in this data?” “Is the reasoning supported with facts?”

Anticipating questions will also reduce the number of interruptions that are bound to happen.

Knowing the answers to these questions with complete clarity helps provide informed answers transforming even unexpected discussions into productive ones, and leaving a positive impression.

Pro tip – At every sentence or illustration in the slide, pause to ask “so what?” That’s a recurring question that senior-level executives ask themselves as well as the presenter. Bake the answers into the presentation for every “so what?” and you have a comprehensive, convincing deck.

Prepare the Short and Long Versions

Even if the slot for the meeting says “12:30-1:30” senior-level meetings are very unpredictable. Five minutes into the meeting, the key decision-maker could get called away on urgent matters. Although the meeting might continue as planned with the rest of the team, the senior executive would want to go through the deck.

Prepare a scan-friendly version, which packs the most important information in less than ten slides. Highlight a ‘takeaway’ at the end of each slide summarizing the information on it. This is the short version. The longer, more detailed document should have more data that supports the goal of the presentation and beef up the benefits. This is the drill-down data the executive will reach for when the shorter version has suitably convinced them.

Pro tip – Try talking to other members in the senior leader’s team to determine their preferences and understand particular pain points. Knowing these will keep the presentation tailored to the last detail.

End With a Persuasive Summary

A C-level presentation does not end merely with Q&As to be forgotten after everybody disperses. Senior leaders are always looking for results, and at the end of the presentation, they need to know what’s next. They need practical call-to-action points they can implement.

The information in the deck should fall into three primary buckets – objectives, benefits, and next steps. The summary gives a composite picture of all the three sections with emphasis on what the audience should do.

“Presentations are about objectives, benefits, and actions.” says Intel’s former Chief Learning Officer to FastCompany

Ending powerfully with repeated emphasis on the benefits prompts the audience to take concrete steps, which delivers the results that leaders seek.

Pro tip – Draw examples from their business history and give solutions by comparing the effectiveness to their biggest competitors. Shape the presentation to ‘speak’ to the specific crowd you are addressing by knowing their personality traits. In effect, this presentation is like an exclusive book that makes sense only to your specific audience.

After that stressful experience the first time, I worked on numerous senior leadership presentations in the last ten years. With each one, I learnt something new and gained more confidence. Today, I find designing C-level presentations an exciting challenge. But more than anything, there’s something deeply satisfying in the knowledge that I am making not just my clients happy but their customers too. Like a double scoop of ice cream!

Nitin Mahajan is an entrepreneur, strategist, and presentation consultant. He is the voice behind this website and, of course, Qcept. Nitin lives and breathes presentations and his mission is to help businesses grow through quality communication and content.

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Data & Finance for Work & Life

Management Presentation: 8 Tips, Examples & a Template

In a corporate context, presenting works wonders for a career. Most professionals get exposure to presenting to informed colleagues and department managers. It’s an ideal way to get visibility and show value. But a management presentation to senior executives who aren’t familiar department nuances is a different ballgame.

A management presentation is a high-level summary to senior executive that optimizes reports to include only the details relevant to directorial decisions . They are notoriously difficult to navigate for two reasons: 1. most executives do not have working knowledge of the nuances in each department , 2. presenters rarely have time to understand executives’ preferences .

More than anything else, good management presenters learn how to strike a balance in the degree of detail: they provide enough detail so executives make informed decisions, but not so much detail that they cause confusion.

This article explores how to make a good management presentations in PowerPoint using 4 management presenting best practices , 4 management presenting techniques , providing examples for each, and finishing with a management presentation template you can apply in real life. You can use it as a jumping off point for deeper communication curriculum .

5 management presenting best practices are:

• Ask what managers prefer ahead of time.
• Have 1 message, and 1 message Only.
• The only words should be “Thought Starters.”
• Keep it short.
• Practice 7 times in advance.

4 management presenting techniques are:

• Use a CSP model – Challenge, Solution, Progress.
• Begin with a summary of exactly 3 points.
• Use only these 3 chart types: bar, line, scatter.
• Design slides with the company logo.

I will use a financial analyst perspective in this article, but everything here applies to data and business analysts as well.

Ask Executives Their Preference Ahead of Time

If you’ve ever taken a class on presentation techniques, you’ve heard the old adage “know your audience.” It’s true, the best way to deliver a great presentation is to align your message with what your audience already understands. The same applies to a management presentation.

The challenge is that, more often than not, executives are too busy for you to get to know them well. This means you hardly get the chance to understand how they like presentations. So what can you do? Well, ask them! There’s no harm in sending an email to understand better. And what’s more, once you know, you can always defer to their preferences in the future.

For a financial management presentation, common questions to ask include the following:

• Do you prefer to see raw data, or only visualizations?
• Do you prefer charts or table summaries?
• Would you like a written explanation on paper for each slide?
• Do you like averages alone, or do you prefer means, or standard deviation?
• What interests you most in a presentation?

If you gather some helpful insights, then your presentation will be that much better. That said, you may not get a response, or it may be quick and not insightful. But most senior executives will appreciate you asking .

The best part is you will be able to surprise them. Using the best practices and techniques below, in additional to any insights gathered form your email, will work wonders for you.

Have 1 Message, and 1 Message Only

The easiest mistake to make on a management presentation is trying to deliver multiple messages. Senior executives go through loads of meetings every day, and each meeting they have includes a wave of information. Your mission should be to deliver 1 essential message so they can easily understand and compartmentalize it.

This is no easy task. When I try to narrow down the focus of my management presentation message, it seems like I leave out critical information along the way. The key is to tell a story to incorporate critical information as part of a story towards the essential message.

For example, imagine you work for a wholesale watch company called Batch Watch . You want to explain a financing operation in which the company has the option of two loans to fund the initial costs of 10,000 watches. These loans have different interest rates and maturity dates. Loan A is better if the company expects to sell the watches within 3 months, while Loan B is better if the company expects to sell over more than 3 months. Each has cancellation fees and cash flow impacts.

Instead of showing the cancellation fees and cash flow impact of the each loan, all you need to say is “ we expect the company to sell them within 3 months, and we recommend loan A for that reason.” If the executives disagree on the sale timeline, they will ask for more information.

This is how you keep senior executives engaged, by integrating them in the story you tell. Ultimately, the essential message of your presentation should be how much profit the company will make from the watch funding operation. Senior executives should leave feeling like the project is in good hands with you, and they only feel that way when you tell a story around the essential message .

Whatever the Message, Use Data

Whatever message you want to send, it needs to be backed up by data. In the example above the data was financial, but it’s not always that simple. Context may require you to provide KPIs and perform extensive data analysis that culminates in a small output that your viewers can easily digest.

You need to be strong with data to deliver a good management presentation. To get started or refresh your memory, you can read AnalystAnswers’ free Intro to Data Analysis eBook .

The Only Words Should be “Thought Starters”

As a general presentation principle, you should not write many thoughts down on presentation slides. Words have two negative impacts on the audience: they demand energy from the reader, and they make the reader feel compelled to read, lest they misunderstand.

If you can avoid putting text blocks altogether, do. If you don’t need any writing at all, don’t. However, if you need guidance as you speak or want to provide reminders for a later data, use “Thought Starters.”

Thought starters are phrases of 3 words maximum that contain ideas leading to the essential message. People often call them “bullet points,” which is common for list-style thought starters. Personally, I prefer to place thought starters at different places on a slide. When I use a chart, for example, I put thought starters at relevant places on the slide.

Keep it Short

Your presentation should never consume more than 80% of the allotted timeframe. This means that if you plan a 5 minutes meeting, deliver the presentation in 4 minutes. If you’re given 30 minutes, do it in 25 minutes. If you have 1 hour, do it in 45 minutes.

By keeping the presentation short, you relieve the audience and you allow for some question buffer. Have you ever sat in a meeting planned for 1 hour, and at 45m it ends early? It’s a pleasure for everyone. Most of us feel like we’re running behind — when you put us ahead of schedule, we love you!

At the same time, senior executives may bombard you with questions throughout the presentation. If you planned to fill the whole timeframe, you won’t finish. But if you planned to finish early, you still have a chance.

And if you use the rest of these best practices and techniques, those senior executives shouldn’t need to ask too many questions!

Practice 7 Times in Advance

There’s a mix of opinions on the number of times you should rehearse a presentation before doing it live, but most people agree that it’s somewhere between 5 and 10 times. If you take nothing else from this article, take this. To deliver a good presentation, prepare excellent slides; to deliver a great presentation, practice presenting them 7 times.

To deliver a good presentation, prepare excellent slides; to deliver a great presentation, practice presenting them 7 times. AnalystAnswers.com

But just practicing isn’t enough, there are a few criteria you must meet:

• Practice in the room you will present in. There’s something about envisioning yourself live that really makes a difference. When you practice in a space other that where you’ll present, it’s good. But when you practice in the “live” room, you’re able to sensitize yourself to the environment, which calms nerves so you can focus on the message.
• Have an audience. We all behave differently when there’s stimulus of other people around. Whenever possible, get one or two people to whom you can present. In addition to getting used to having an audience, you’ll also get some feedback.
• Use the same volume of voice. When we’re not “live,” we have a tendency to hold back on our voice. This is detrimental to the presentation because you feel taken off guard by your own voice. Make sure to envision yourself in front of the senior execs when you practice.

Best Practices Recap

We’ve addressed 5 best practices — now let’s turn our attention to 4 specific techniques you can easily implement. And when you do, that work wonders for management presenting.

Use a CSP Model (Challenge, Solution, Progress)

Every presentation needs structure, but it’s easy to forget that we need to guide our audience. A great way to structure management reports is using the CSP model. CSP stands for Challenge, Solution, Progress, and it’s exactly what it sounds like.

You need to explain the challenge or goal, explain what the solution to the challenge is (or how to achieve the goal), and show where you are in the steps to completing that goal.

For example, let’s look at our Batch Watch case. Imagine you need to find funding for a new product launch — $100,000 to be exact. A sample CSP model for this would be a slide that shows:

By using the CSP model, you guide the audience. However, it’s important to note that the CSP model is not a summary . It’s an overview of the process, but a summary should always come before. Let’s talk about it now.

Begin with a Summary of Exactly 3 Points

Any good presentation begins with a summary. And a good summary communicates the essential message simply in 3 points. However, the summary is not the same thing as the CSP model. Instead, it provides an alternative view on the challenge and and solution.

For example, using our Batch Watch case of funding a new product, you could address a summary in the following way:

• Challenge, Solution, Progress
• Funding acquisition
• Project Timeline

This provides additional details that are most relevant to the project and carry added value to the CSP model.

Use only Bar Charts (aka Column Charts), Line Graphs, and Scatter Plots

Whether it’s for data, financial, and business analyst topics , management presentations should only ever have bar charts, line graphs, and scatter plots. They are common, rich in information, and well understood. Any other kind of graph is distracting more than anything else.

A bar graph is useful when you want to compare like variables. For example, if you want to show the average size of Canadian trout versus American trout. A common mistake, though, is to use bar graphs to show change over time. While it’s not incorrect to do so, line graphs are better for this purpose.

A line graph is useful when you want to show change in one variable over time (we call this time series data). For example, if you want to show the progression of revenues over time, line graphs are the perfect way to do so.

A scatter plot is best when you want to compare a set of observations of one variable to a set of observations of another. It’s the ideal way to quickly visualize the relationship between two variables. For example, if you want to see how company revenues compare to GDP, you could use a scatter plot like this:

For example, let’s look at our Batch Watch case. If we want to see how our company is performing compared to the economy as a whole, we could use this scatter plot. As you can see, we have a positive (bottom left to top right) relationship, but a weak one (points not clustered closely).

Design Slides Using the Company Logo

When you’re presenting to senior executives, you want your slides to look professional. The best way to do that is by putting your company logo on them, including any corporate design standards (colors, fonts, etc). Show through your presentation that you belong to the same company, and that you’re in it in spirit. For example, let’s add the AnalystAnswers.com logo to our CSP slide:

Techniques Recap

Here’s a sample management presentation template below. I hope you understand after reading this article that management presentation is more about your delivery than it is about the slides you prepare.

Download Management Presentation Template for Free

While the techniques we’ve discussed will help you build a good presentation, your success really depends on how well you deliver the ideas needed to help senior executives make decisions. At the end of the day, it’s all about balance.

If you only remember two things from this article, remember that great management presenters give enough detail to inform senior executive but not too much that they cause confusion, and great management presenters make sure they do so by practicing 7 times in advance. You’ll have to practice, practice, practice.

About the Author

Noah is the founder & Editor-in-Chief at AnalystAnswers. He is a transatlantic professional and entrepreneur with 5+ years of corporate finance and data analytics experience, as well as 3+ years in consumer financial products and business software. He started AnalystAnswers to provide aspiring professionals with accessible explanations of otherwise dense finance and data concepts. Noah believes everyone can benefit from an analytical mindset in growing digital world. When he's not busy at work, Noah likes to explore new European cities, exercise, and spend time with friends and family.

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How to Create Slides That Suit Your Superiors: 11 Tips

When you’re pitching ideas or budgets to execs in your organization, you need to deliver slides that fit those particular people just right. This checklist identifies the key considerations.

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I recently interviewed 20 of my customers, all in senior roles at Fortune 100 companies, and asked them their biggest pain point in presenting to higher-ups and even colleagues. What I heard consistently was that it can feel like Goldilocks bouncing from one option to the next, testing to figure out what’s “just right.” Does the audience want deep reports? Sparse slides? Something in between? Like … what?

Teams often come to presentation meetings with vast amounts of backup content just in case an exec wants to take a deep dive on any given point. There’s often a struggle to anticipate every direction attendees might want to go. It’s frustrating, and it’s not efficient.

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There are many ways to build slides. I’m not just talking about crafting them well versus poorly. I’m talking about all of the important decisions regarding how to organize them, how much text to use, when to lean into a chart, the best ways to use bullets and color, and whether to include an appendix with additional information. Before you make your next proposal or request of the executive team, use this list of 11 tips for your next set of slides as a guide.

Four Things You Must Have in Every Exec’s Slides

Before we drill down into the harder aspects, the ones where your executives’ tastes may vary widely, let’s quickly cover four aspects that you can consider the building blocks — the basics you should never proceed without.

Start with an executive summary. Begin the slide deck with a tight executive summary that follows a three-act structure. First, start with stating the current realities. Second, clearly state the problem or opportunity your idea addresses and its potential impact. Third, explain how your recommendation solves the problem or exploits the opportunity and the next steps you’re proposing.

Have a logical organization. The arc of the deck — the package from beginning to end — should make sense. If your audience reads only the headline of every slide, the order should be coherent and make most of the case for you. The content below each slide’s headline must support the statement made in the title. Remove everything that doesn’t support your point; as writers will tell you, you sometimes need to “kill your darlings” when you’re editing.

Begin the slide deck with a tight executive summary that follows a three-act structure.

Make it skimmable. Help your audience to quickly grasp the point without getting bogged down in details. Create a clear visual hierarchy. Guide the reader’s eye through the content: Use bold headings, bullet points, and numbered lists to break down information into digestible pieces. Highlight key takeaways or conclusions in a different color or font size to draw attention to these critical points.

Focus on concise insights. Succinct statements with clear insights are everyone’s jam. Every slide should serve a purpose and contribute directly to the decision-making process. Distill complex information. Don’t use 100 words when 20 words will nail it. If you’re having difficulty trimming, consider using company-approved AI tools to help you take out the fluff.

Five Preferences to Confirm With the Person You Want to Reach

Now we’ll delve into what your particular audience does and does not want. If you haven’t yet, start by asking the person you’re presenting to what they generally prefer. They probably know themselves well but have not been asked to articulate how they like to receive information.

Ask how dense is too dense. Some executives prefer detailed slides with comprehensive data. Others favor a more high-level approach. You’re weighing how to balance informative content with readability, ensuring that slides are not overloaded yet are sufficiently detailed to support decision-making.

Confirm the delivery format and timing. Some execs like information presented to them. Others prefer a pre-read of the material followed by a discussion. I always recommend our tool Slidedocs (I’ve written a free e-book on them), which are visual documents using both words and images. The templates help presenters organize their thoughts into a document for a pre-read or a read-along. They are designed to be skimmable and able to travel through your organization without the help of a presenter.

I’m a huge fan of pre-reads and prefer to use my time in meetings to ask questions and build alignment. If your audience didn’t review your material in advance, ask at the top of the meeting whether they would like you to present it or would prefer to read through it and then discuss it.

Find out how much data visualization they prefer. Charts, graphs, photos, and illustrations often communicate complex data more clearly than words alone. When execs can see what you’re saying, they often can better understand the impact of your idea. Does the exec want to understand exact numbers? Bar charts allow them to move their eyes across a series of specifics. Does the exec want to know the shape of a trend over time? Line charts can show the pattern. (See “Classic Charts Communicate Data Quickly.”) Some prefer charts with annotations that draw attention to what you think is the most important point. Others want to make their own conclusions from the data.

One of my clients, the CEO of a massive commercial real estate company, doesn’t want anything visualized. He prefers numbers, only in a table, and only in two colors — black and red. You might think this is archaic. But the fact that he’s clear to his teams about what he wants takes all the mystery out of how to communicate with him.

When the stakes are high, have a conceptual thinker help with diagrams and concepts. If you don’t have one on your team, and when it’s high stakes, find an internal designer to help you or hire one. You can’t afford to have the baby (your idea) thrown out with the bathwater (terrible slides).

Identify which details need spelling out. How well do the people you’re presenting to know the landscape and function of the company and products you’re talking about? For example, if your engineering team threw a slide into a deck about an issue that requires executive approval, do the execs all speak geek? Or do you need to explain the technology so that they will really understand the ask? Either eliminate internal jargon and acronyms or unpack those bits, especially if your proposal deeply involves expertise outside of the executives’ domain.

Ask whether appendices will be useful. When you’re organizing a presentation, you often troll data, read through complicated reports, and even hire external experts to figure out what’s best for the company. Do your execs want access to that supporting data? You can add a document to the end of the presentation as an appendix to show all of the data and source material. This allows the main content of the slides to remain focused and accessible while still providing comprehensive background information for those who want more.

Two Tips to Improve Your Presentation Skills

Getting materials in place is the biggest step. They will be your best tools for selling your ideas. But there are two extra areas to pay attention to as a presenter: how you handle questions and how you use every experience to improve.

Anticipate questions, and practice your answers. Before you have your meeting, gather a small team to challenge every point you make. Invite colleagues you trust to role-play as “a rapidly inquisitive exec” or “the doubting naysayer exec” so you are prepared to present your idea well. They’re gonna grill you, and practicing will help you remain unruffled when it happens.

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Ask for feedback after the presentation. Establish a feedback loop with those you presented to. Ask what worked well and how you can improve. If attendees don’t have the time, find people who have had their ideas funded and talk to them about what they did that worked. Advice and some perspective will help you nail your performance even better next time.

Empathetically understanding your audience members and how they process information, whether it’s executives or peers, sets up your ideas for success. Clarity creates efficiency. When a presentation fits just right, you’ve given your great thinking the best chance of moving through your organization and having maximum impact.

About the Author

Nancy Duarte is CEO of Duarte Inc. , a communication company in the Silicon Valley. She’s the author of six books, including DataStory: Explain Data and Inspire Action Through Story (Ideapress Publishing, 2019).

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20 Great Examples of PowerPoint Presentation Design [+ Templates]

Published: January 17, 2024

When it comes to PowerPoint presentation design, there's no shortage of avenues you can take.

While all that choice — colors, formats, visuals, fonts — can feel liberating, it‘s important that you’re careful in your selection as not all design combinations add up to success.

In this blog post, I’m sharing some of my favorite PowerPoint tips and templates to help you nail your next presentation.

Table of Contents

What makes a good PowerPoint presentation?

Powerpoint design ideas, best powerpoint presentation slides, good examples of powerpoint presentation design.

In my opinion, a great PowerPoint presentation gets the point across succinctly while using a design that doesn't detract from it.

Here are some of the elements I like to keep in mind when I’m building my own.

1. Minimal Animations and Transitions

Believe it or not, animations and transitions can take away from your PowerPoint presentation. Why? Well, they distract from the content you worked so hard on.

A good PowerPoint presentation keeps the focus on your argument by keeping animations and transitions to a minimum. I suggest using them tastefully and sparingly to emphasize a point or bring attention to a certain part of an image.

2. Cohesive Color Palette

I like to refresh my memory on color theory when creating a new PowerPoint presentation.

A cohesive color palette uses complementary and analogous colors to draw the audience’s attention and help emphasize certain aspects at the right time.

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It‘s impossible for me to tell you the specific design ideas you should go after in your next PowerPoint, because, well, I don’t know what the goal of your presentation is.

Luckily, new versions of PowerPoint actually suggest ideas for you based on the content you're presenting. This can help you keep up with the latest trends in presentation design .

PowerPoint is filled with interesting boilerplate designs you can start with. To find these suggestions, open PowerPoint and click the “Design” tab in your top navigation bar. Then, on the far right side, you'll see the following choices:

1. “The Search for Meaning in B2B Marketing,” Velocity Partners

We've said it once , and I'll say it again: I love this presentation from Velocity Partner's Co-Founder Doug Kessler. Not only is the content remarkable, but the design is also quite clever.

What I like: While each slide employs the same background visual, the copy in the notebook unfolds brilliantly through a series of colorful doodles and bold text. This gives the presentation a personal feel, which aligns with the self-reflective nature of the concept.

2. “You Don't Suck at PowerPoint,” Jesse Desjardins

If the contrast used throughout this PowerPoint presentation design were a human, I'd marry it.

This skillful presentation from Jesse Desjardins employs the perfect color palette : balancing black and white photos with pops of fluorescent pink, yellow, and blue.

What I like: The cheeky vintage photos work to reinforce the copy on each slide, making the presentation both interesting and visually appealing.

3. “Accelerating Innovation in Energy,” Accenture

Balancing visual backgrounds with text isn't easy. More often than not, the text is formatted in a way that winds up getting lost in the image. Not for Accenture.

What I like: This presentation combated this issue by combining shapes and graphics to create contrast between the text and the background. Well done.

4. “Visual Design with Data,” Seth Familian

Whenever I’m tasked with presenting a lot of information in a little bit of time, things can get sort of messy. To simplify this type of presentation, I like to use a visual agenda like the one shown above.

What I like: This index clearly signifies the start and finish of each section to make it easier for the viewer to follow along. The presenter takes it further by including an additional agenda for each exercise, so that the audience knows exactly what they're supposed to do.

5. “How to Craft Your Company's Storytelling Voice,” MarketingProfs

Do you love these hand-drawn illustrations or do you love these hand-drawn illustrations? I mean, c'mon, this presentation by MarketingProf is amazing.

What I like: Certainly, it would have been easier to generate these designs online, but this approach highlights their commitment to creating an out-of-the-box piece of content. And as a result, this presentation stands out in the best way possible.

6. “Blitzscaling: Book Trailer,” Reid Hoffman

If you're going to go the minimalistic route, I’d take note of this PowerPoint presentation example from Reid Hoffman.

This clean design adheres to a simple, consistent color scheme with clean graphics peppered throughout to make the slides more visually interesting.

What I like: Overall there are no frills or unnecessary additions, which allows the informative content to take priority.

7. “Healthcare Napkins,” Dan Roam

This presentation dates back to 2009, but the design is still as good as ever. The colorful, quirky doodles help tell the story while also serving as an interesting way to illustrate data (see slides 20 and 21).

What I like: For visual learners, this approach is much more inviting than a series of slides riddled with text-heavy bullet points.

8. “One Can Be Diverse: An Essay on Diversity,” With Company

This presentation employs both powerful images and modern typography to illustrate the point.

What I like: While many of the slides contain long quotes, they are broken up in a way that makes them easily digestible. Not to mention all of the text is crisp, clean, and concise.

9. " 10 Things Your Audience Hates About Your Presentation ," Stinson

This simplistic presentation example employs several different colors and font weights, but instead of coming off as disconnected, the varied colors work with one another to create contrast and call out specific concepts.

What I like: The big, bold numbers help set the reader's expectations, as they clearly signify how far along the viewer is in the list of tips.

10. “Pixar's 22 Rules to Phenomenal Storytelling,” Gavin McMahon

This presentation by Gavin McMahon features color in all the right places. While each of the background images boasts a bright, spotlight-like design, all the characters are intentionally blacked out.

What I like: This helps keep the focus on the tips, while still incorporating visuals. Not to mention, it's still easy for me to identify each character without the details. (I found you on slide eight, Nemo.)

11. “Facebook Engagement and Activity Report,” We Are Social

Here's another great example of data visualization in the wild.

What I like: Rather than displaying numbers and statistics straight up, this presentation calls upon interesting, colorful graphs, and charts to present the information in a way that just makes sense.

12. “The GaryVee Content Model,” Gary Vaynerchuk

This wouldn‘t be a true Gary Vaynerchuk presentation if it wasn’t a little loud, am I right?

What I like: Aside from the fact that I love the eye-catching, bright yellow background, Vaynerchuk does a great job of incorporating screenshots on each slide to create a visual tutorial that coincides with the tips. He also does a great job including a visual table of contents that shows your progress as you go .

13. “20 Tweetable Quotes to Inspire Marketing & Design Creative Genius,” IMPACT Branding & Design

We‘ve all seen our fair share of quote-chronicling presentations but that isn’t to say they were all done well. Often the background images are poor quality, the text is too small, or there isn't enough contrast.

Well, this professional presentation from IMPACT Branding & Design suffers from none of said challenges.

What I like: The colorful filters over each background image create just enough contrast for the quotes to stand out.

14. “The Great State of Design,” Stacy Kvernmo

This presentation offers up a lot of information in a way that doesn't feel overwhelming.

What I like: The contrasting colors create visual interest and “pop,” and the comic images (slides 6 through 12) are used to make the information seem less buttoned-up and overwhelming.

15. “Clickbait: A Guide To Writing Un-Ignorable Headlines,” Ethos3

Not going to lie, it was the title that convinced me to click through to this presentation but the awesome design kept me there once I arrived.

What I like: This simple design adheres to a consistent color pattern and leverages bullet points and varied fonts to break up the text nicely.

16. “Digital Transformation in 50 Soundbites,” Julie Dodd

This design highlights a great alternative to the “text-over-image” display we've grown used to seeing.

What I like: By leveraging a split-screen approach to each presentation slide, Julie Dodd was able to serve up a clean, legible quote without sacrificing the power of a strong visual.

17. “Fix Your Really Bad PowerPoint,” Slide Comet

When you‘re creating a PowerPoint about how everyone’s PowerPoints stink, yours had better be terrific. The one above, based on the ebook by Seth Godin, keeps it simple without boring its audience.

What I like: Its clever combinations of fonts, together with consistent color across each slide, ensure you're neither overwhelmed nor unengaged.

18. “How Google Works,” Eric Schmidt

Simple, clever doodles tell the story of Google in a fun and creative way. This presentation reads almost like a storybook, making it easy to move from one slide to the next.

What I like: This uncluttered approach provides viewers with an easy-to-understand explanation of a complicated topic.

19. “What Really Differentiates the Best Content Marketers From The Rest,” Ross Simmonds

Let‘s be honest: These graphics are hard not to love. I especially appreciate the author’s cartoonified self-portrait that closes out the presentation. Well played, Ross Simmonds.

What I like: Rather than employing the same old stock photos, this unique design serves as a refreshing way to present information that's both valuable and fun.

20. “Be A Great Product Leader,” Adam Nash

This presentation by Adam Nash immediately draws attention by putting the company's logo first — a great move if your company is well known.

What I like: He uses popular images, such as ones of Megatron and Pinocchio, to drive his points home. In the same way, you can take advantage of popular images and media to keep your audience engaged.

PowerPoint Presentation Examples for the Best Slide Presentation

Mastering a PowerPoint presentation begins with the design itself.

Get inspired by my ideas above to create a presentation that engages your audience, builds upon your point, and helps you generate leads for your brand.

Editor's note: This post was originally published in March 2013 and has been updated for comprehensiveness. This article was written by a human, but our team uses AI in our editorial process. Check out our full disclosure to learn more about how we use AI.

Don't forget to share this post!

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Do You Have What It Takes to Give a Great Presentation?

• Michael Foley

Three skills anyone can learn.

Great presentations are all about how well you engage your audience. Here are three ways to do that:

• Be balanced: Strike the right balance between structure and improvisation during your presentation. Rather than preparing your content in a narrative form, take the time to segment it by topic. Leave room to respond to your audience’s immediate needs, even if that means going off script for a while.
• Be generous: You goal should be to offer your audience something of value. Let this  principle guide the content you choose to present.
• Be human: Rehearse to familiarize yourself with your content, but do not memorize it. If you go blank for a moment, don’t apologize and act flustered, just pause, take a breath, collect your thoughts and continue on. Your audience wants you to be relatable.

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Have you ever seen a really great presentation? Honestly, they’re pretty rare.

• Michael Foley is an educator and coach, and founder of Clarity Centra l, a communications training and consulting firm based in Chicago. He is also an adjunct lecturer of leadership at the Kellogg Graduate School of Management at Northwestern University.

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Home Blog Business Executive Summary: A Guide to Writing and Presentation

Executive Summary: A Guide to Writing and Presentation

Executive summaries precede nearly every type of business document. Despite being the shortest part, they often leave the biggest impression on the reader. Yet, many writers choose to treat an executive summary as an afterthought. (And some presenters too!). Why? Because writing an executive summary is a seemingly hard task. But our mission is to prove otherwise! 

What is an Executive Summary?

An executive summary is a preface to a larger business document such as an annual report, business plan, or whitepaper, succinctly summarizing the key discussion points. Effectively, an executive summary offers a preview of the content, so that the reader could form a baseline opinion about the contents prior to diving into a deep reading session. 

The University of Arizona offers a more elaborated executive summary definition which also notes that an executive summary should:

• Restate the purpose of the follow-up document
• Highlight the key discussion points and most notable facts 
• Relay any notable results, conclusions, or recommendations

Though an executive summary is just a foreword to a bigger report, it’s one of the most labor-intensive items as you have to condense a lot of information into a high-level summary. Oftentimes, an executive summary also gets prominent placement in the follow-up presentation, done on the report.  

Executive Summary Examples

Nearly every type of business document will have an executive summary. Some are better structured and presented than others. But it’s not just limited to business documents. Executive summaries are also used in scientific projects, articles, and education. Below are several admirable executive summary examples you may want to use as an inspiration for writing. 

Accenture: Gaming: The Next Super Platform 

This executive summary for an industry report opens with some big quantifiable claims, clearly communicating the main agenda — describing the size and state of the global gaming market. The gaming industry is a huge market. The pullout texts on the sidebar further detail the scope of the document. Plus clarify for whom this report is intended. 

IBM: Cost of a Data Breach Report 2020 

IBM conducts an annual joint report on cybersecurity with Ponemon Institute. They open the executive summary with a brief recap of their mission and past research. Then dwell on this year’s findings and methodology. If you are writing an executive summary for a similarly massive original research, it’s worth focusing more on your techniques for obtaining data and arriving at the conclusions as IBM did. 

Deloitte Digital: Exploring the value of emotion-driven engagement

Deloitte selected a more narrative style for this executive summary, mixing some key data points and methodology with the core messaging of the report. This is a good example of structured data presentation . On one hand, you have an engaging narration flow. On the other, the summary covers all the important discussion points. 

Executive Summary Format

As the above executive summary examples illustrated, there is no one fit-it-all format for writing an executive study. The best approach depends on your report type, purpose, and contents. 

That being said, an executive summary needs to fulfill several earlier mentioned criteria — offer a preview, provide key information at glance, showcase any results, recommendations. That’s what most readers expect to see on the first page after all. 

The easiest way to approach writing is to draft a preliminary executive summary outline featuring the following subsections:

• General introduction, explaining the key problems discussed 
• Main problem statement(s)
• Selected findings or recommendations 
• The importance of discussed points 

Since you’d also be likely working on presenting the executive summary to other stakeholders , it helps you keep the above structured as bullet points at first. So that you could easily transfer the main ideas to your executive summary PowerPoint slide . 

How Long Should an Executive Summary Be?

As a rule of thumb, an executive summary should not go longer than one vertical page. That is an equivalent of 300-500 words, depending on the typeface. For longer reports, two pages (a horizontal split) may be acceptable. But remember, brevity is key. You are working on a trailer for a movie (the full report). 

How to Write an Executive Summary: a 3-Step Framework 

You can start with the aforementioned loose format and then adapt it to your document type. Remember, you don’t need to follow all the recommendations to a T. Instead, mix some ideas to make your executive summary sound both professional and engaging. Here are several tips for that:  

1. Start with a Problem Statement 

Think of the first paragraph as if of an opening slide for a presentation : you need to make a big compelling statement that immediately communicates your agenda. Set the scene for the reader. There are several ways to do so:

• Answer the “why now” question in the opening paragraph 
• Address the urgency of the matter 
• Highlight the importance of the discussed issue 

Alternatively, you can also go for a more traditional opening and explain the background of the research and discussed issue. For example, if you have conducted a go-to-market strategy evaluation for the team you can start by saying that “This report analyzed online furniture brand performance in 5 target EMEA markets in terms of market share, local brand recall, brand preference, and estimated online sales volumes.” Afterward, briefly communicate the main aim of the report. 

2. Present the Main Discussion Points 

Next, flesh out what’s included in the scope of this report to properly manage the reader’s expectations. You can use the report’s section subheads as key discussion points or come up with snappier, more descriptive statements. 

Here are several good writing practices to follow: 

• Use bullet points and numbered lists to break down text blocks. 
• Quantify the biggest findings when possible. Style them as “call-outs”. 
• Mention the limitations of your report and what it does not account for. 
• Discuss the used research methods and data sources. 

Finally, summarize the findings in one concluding paragraph if you have space. Or style it as a featured quote to draw the reader’s eye towards crucial information. 

3. List the Recommendations or Next Steps 

The bottom part of the page, around 100-150 words should be allocated towards underlining the results, conclusions, and follow-up action expected from the reader. Summarize what you have found during the course of your research. Mention if you have identified any specific type of solution or a type of recommended action. 

Once you are done, send over an executive summary draft to a team member who hasn’t seen the complete report. Ask for their feedback. Can they tell what the report content is after reading the summary? Does the summary intrigue them? Is it descriptive enough for someone without any other context into the matter? Use the critique to further improve the document. 

How to Prepare an Executive Summary Presentation 

High chances are that you’ll also be asked to write the copy for the executive summary presentation, and perhaps even design it too. So let’s get you up to speed on this aspect as well.

How Does an Executive Summary Slide Look Like in PPT?

There’s no ultimate look for an executive summary slide as most presenters customize it to best reflect the content they’d want to showcase. But if you want some universal example, here’s our executive summary slide template : 

You can build an entire slide deck tailored for an executive summary or business presentation by using our AI Presentation Maker . Fill the topic, analyze & edit the proposed outline, and select a design. That’s it! You can create an engaging executive summary slide deck with any number of slides.

What Makes a Good Executive Summary Slide?

A good executive summary slide visually communicates all the important information from the full report. Typically, it’s an even more condensed version of the written executive summary, prefacing the document. Thus to create a good executive summary slide, be prepared to do some ruthless editing.

Include a condensed version of the: 

• Main problem statement or report agenda 
• Key findings. Prioritize quantifiable ones
• Recommendations and next steps.

Also, you will need some PowerPoint design mastery to ensure that an executive summary in your PowerPoint presentation looks compelling, but not cluttered. Prioritize white space. Here is where a good executive summary template can make your life easier. To minimize the number of texts, add icons and other simple visualizations. Trim headers and subheads to give the slide even more breathing room.

For those looking to create an engaging and visually appealing presentation, consider utilizing professional presentation templates to enhance the visuals of your executive summary slide. These templates are specifically designed to help presenters convey their message effectively and with style, ensuring that your audience remains captivated and fully understands the key points of your report.

How to Write an Executive Summary for a Presentation

Most likely you won’t need to write a brand new copy for this slide, but rather adapt the text at hand. That already makes your job a lot easier when summarizing a presentation into an executive summary slide. Still, you don’t want to mess anything up. So stick with the executive summary template you’ve chosen and fill in the gaps using our tips. 

1. Keep the Tone Consistent 

Use the same tone of voice and word choices in your slide deck as you’ve adopted in the report. If the tone of your presentation speech differs too much with terms used on the slide and in the report copy, some audience members may get confused, and then disengaged. 

2. Focus on Telling a Story 

Stakeholders will have the extra time to read the “dry” report. During the presentation, your main goal is to draw their attention to the most important issue, showcase the value-packed inside the report, and make them eager to learn more by actually flipping the full copy afterward. 

3. Chop Full Sentences into Bullet Points 

Go snappy and present information in a snackable manner. Remember, our brain can only keep 3-5 items at once in the working memory. So you shouldn’t try to overload the audience with a long list of “very important points” in one sitting.  

Also, per a recent presentation survey, among the 3 things that annoy audiences most about presentations are slides that include full sentences of text. So, when working on your presentation summary slide, trim those lengthy texts and move on some of the other points to separate slides. 

4. Don’t Go Data Galore 

Including numbers and data visualizations is a great way to present your executive summary. However, overloading your data slides with data nuggets makes your presentation less impactful. 

As presentation design expert Nancy Duarte explains : 

“Data slides aren’t really about the data. They’re about the meaning of the data. It’s up to you to make that meaning clear before you click away. Otherwise, the audience won’t process — let alone buy — your argument.”

It’s a good idea to spotlight 3 main data points on your executive summary slide. Then use some extra minutes to comment on why you’ve chosen to present these. 

To Conclude

An executive summary is the first page and/or slide a reader will see. That’s why the stakes are high to make it look just right. Granted, that shouldn’t be an issue. Since you now know how to write, design, and present a compelling executive summary to others! 

1. Project Summary PowerPoint Template

Use This Template

2. Simple Executive Summary Slide Template for PowerPoint

3. One Page Strategy Summary PowerPoint Template

4. Executive Summary PowerPoint Template

5. Executive Business PowerPoint Template

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There is unequivocal evidence that Earth is warming at an unprecedented rate. Human activity is the principal cause.

• While Earth’s climate has changed throughout its history , the current warming is happening at a rate not seen in the past 10,000 years.
• According to the Intergovernmental Panel on Climate Change ( IPCC ), "Since systematic scientific assessments began in the 1970s, the influence of human activity on the warming of the climate system has evolved from theory to established fact." 1
• Scientific information taken from natural sources (such as ice cores, rocks, and tree rings) and from modern equipment (like satellites and instruments) all show the signs of a changing climate.
• From global temperature rise to melting ice sheets, the evidence of a warming planet abounds.

The rate of change since the mid-20th century is unprecedented over millennia.

Earth's climate has changed throughout history. Just in the last 800,000 years, there have been eight cycles of ice ages and warmer periods, with the end of the last ice age about 11,700 years ago marking the beginning of the modern climate era — and of human civilization. Most of these climate changes are attributed to very small variations in Earth’s orbit that change the amount of solar energy our planet receives.

The current warming trend is different because it is clearly the result of human activities since the mid-1800s, and is proceeding at a rate not seen over many recent millennia. 1 It is undeniable that human activities have produced the atmospheric gases that have trapped more of the Sun’s energy in the Earth system. This extra energy has warmed the atmosphere, ocean, and land, and widespread and rapid changes in the atmosphere, ocean, cryosphere, and biosphere have occurred.

Earth-orbiting satellites and new technologies have helped scientists see the big picture, collecting many different types of information about our planet and its climate all over the world. These data, collected over many years, reveal the signs and patterns of a changing climate.

Scientists demonstrated the heat-trapping nature of carbon dioxide and other gases in the mid-19th century. 2 Many of the science instruments NASA uses to study our climate focus on how these gases affect the movement of infrared radiation through the atmosphere. From the measured impacts of increases in these gases, there is no question that increased greenhouse gas levels warm Earth in response.

Scientific evidence for warming of the climate system is unequivocal.

Intergovernmental Panel on Climate Change

Ice cores drawn from Greenland, Antarctica, and tropical mountain glaciers show that Earth’s climate responds to changes in greenhouse gas levels. Ancient evidence can also be found in tree rings, ocean sediments, coral reefs, and layers of sedimentary rocks. This ancient, or paleoclimate, evidence reveals that current warming is occurring roughly 10 times faster than the average rate of warming after an ice age. Carbon dioxide from human activities is increasing about 250 times faster than it did from natural sources after the last Ice Age. 3

The Evidence for Rapid Climate Change Is Compelling:

Global Temperature Is Rising

The planet's average surface temperature has risen about 2 degrees Fahrenheit (1 degrees Celsius) since the late 19th century, a change driven largely by increased carbon dioxide emissions into the atmosphere and other human activities. 4 Most of the warming occurred in the past 40 years, with the seven most recent years being the warmest. The years 2016 and 2020 are tied for the warmest year on record. 5 Image credit: Ashwin Kumar, Creative Commons Attribution-Share Alike 2.0 Generic.

The Ocean Is Getting Warmer

The ocean has absorbed much of this increased heat, with the top 100 meters (about 328 feet) of ocean showing warming of 0.67 degrees Fahrenheit (0.33 degrees Celsius) since 1969. 6 Earth stores 90% of the extra energy in the ocean. Image credit: Kelsey Roberts/USGS

The Ice Sheets Are Shrinking

The Greenland and Antarctic ice sheets have decreased in mass. Data from NASA's Gravity Recovery and Climate Experiment show Greenland lost an average of 279 billion tons of ice per year between 1993 and 2019, while Antarctica lost about 148 billion tons of ice per year. 7 Image: The Antarctic Peninsula, Credit: NASA

Glaciers Are Retreating

Glaciers are retreating almost everywhere around the world — including in the Alps, Himalayas, Andes, Rockies, Alaska, and Africa. 8 Image: Miles Glacier, Alaska Image credit: NASA

Snow Cover Is Decreasing

Satellite observations reveal that the amount of spring snow cover in the Northern Hemisphere has decreased over the past five decades and the snow is melting earlier. 9 Image credit: NASA/JPL-Caltech

Sea Level Is Rising

Global sea level rose about 8 inches (20 centimeters) in the last century. The rate in the last two decades, however, is nearly double that of the last century and accelerating slightly every year. 10 Image credit: U.S. Army Corps of Engineers Norfolk District

Arctic Sea Ice Is Declining

Both the extent and thickness of Arctic sea ice has declined rapidly over the last several decades. 11 Credit: NASA's Scientific Visualization Studio

Extreme Events Are Increasing in Frequency

The number of record high temperature events in the United States has been increasing, while the number of record low temperature events has been decreasing, since 1950. The U.S. has also witnessed increasing numbers of intense rainfall events. 12 Image credit: Régine Fabri,  CC BY-SA 4.0 , via Wikimedia Commons

Ocean Acidification Is Increasing

Since the beginning of the Industrial Revolution, the acidity of surface ocean waters has increased by about 30%. 13 , 14 This increase is due to humans emitting more carbon dioxide into the atmosphere and hence more being absorbed into the ocean. The ocean has absorbed between 20% and 30% of total anthropogenic carbon dioxide emissions in recent decades (7.2 to 10.8 billion metric tons per year). 1 5 , 16 Image credit: NOAA

1. IPCC Sixth Assessment Report, WGI, Technical Summary . B.D. Santer et.al., “A search for human influences on the thermal structure of the atmosphere.” Nature 382 (04 July 1996): 39-46. https://doi.org/10.1038/382039a0. Gabriele C. Hegerl et al., “Detecting Greenhouse-Gas-Induced Climate Change with an Optimal Fingerprint Method.” Journal of Climate 9 (October 1996): 2281-2306. https://doi.org/10.1175/1520-0442(1996)009<2281:DGGICC>2.0.CO;2. V. Ramaswamy, et al., “Anthropogenic and Natural Influences in the Evolution of Lower Stratospheric Cooling.” Science 311 (24 February 2006): 1138-1141. https://doi.org/10.1126/science.1122587. B.D. Santer et al., “Contributions of Anthropogenic and Natural Forcing to Recent Tropopause Height Changes.” Science 301 (25 July 2003): 479-483. https://doi.org/10.1126/science.1084123. T. Westerhold et al., "An astronomically dated record of Earth’s climate and its predictability over the last 66 million years." Science 369 (11 Sept. 2020): 1383-1387. https://doi.org/10.1126/science.1094123

2. In 1824, Joseph Fourier calculated that an Earth-sized planet, at our distance from the Sun, ought to be much colder. He suggested something in the atmosphere must be acting like an insulating blanket. In 1856, Eunice Foote discovered that blanket, showing that carbon dioxide and water vapor in Earth's atmosphere trap escaping infrared (heat) radiation. In the 1860s, physicist John Tyndall recognized Earth's natural greenhouse effect and suggested that slight changes in the atmospheric composition could bring about climatic variations. In 1896, a seminal paper by Swedish scientist Svante Arrhenius first predicted that changes in atmospheric carbon dioxide levels could substantially alter the surface temperature through the greenhouse effect. In 1938, Guy Callendar connected carbon dioxide increases in Earth’s atmosphere to global warming. In 1941, Milutin Milankovic linked ice ages to Earth’s orbital characteristics. Gilbert Plass formulated the Carbon Dioxide Theory of Climate Change in 1956.

3. IPCC Sixth Assessment Report, WG1, Chapter 2 Vostok ice core data; NOAA Mauna Loa CO2 record O. Gaffney, W. Steffen, "The Anthropocene Equation." The Anthropocene Review 4, issue 1 (April 2017): 53-61. https://doi.org/abs/10.1177/2053019616688022.

4. https://www.ncei.noaa.gov/monitoring https://crudata.uea.ac.uk/cru/data/temperature/ http://data.giss.nasa.gov/gistemp

5. https://www.giss.nasa.gov/research/news/20170118/

6. S. Levitus, J. Antonov, T. Boyer, O Baranova, H. Garcia, R. Locarnini, A. Mishonov, J. Reagan, D. Seidov, E. Yarosh, M. Zweng, " NCEI ocean heat content, temperature anomalies, salinity anomalies, thermosteric sea level anomalies, halosteric sea level anomalies, and total steric sea level anomalies from 1955 to present calculated from in situ oceanographic subsurface profile data (NCEI Accession 0164586), Version 4.4. (2017) NOAA National Centers for Environmental Information. https://www.nodc.noaa.gov/OC5/3M_HEAT_CONTENT/index3.html K. von Schuckmann, L. Cheng, L,. D. Palmer, J. Hansen, C. Tassone, V. Aich, S. Adusumilli, H. Beltrami, H., T. Boyer, F. Cuesta-Valero, D. Desbruyeres, C. Domingues, A. Garcia-Garcia, P. Gentine, J. Gilson, M. Gorfer, L. Haimberger, M. Ishii, M., G. Johnson, R. Killick, B. King, G. Kirchengast, N. Kolodziejczyk, J. Lyman, B. Marzeion, M. Mayer, M. Monier, D. Monselesan, S. Purkey, D. Roemmich, A. Schweiger, S. Seneviratne, A. Shepherd, D. Slater, A. Steiner, F. Straneo, M.L. Timmermans, S. Wijffels. "Heat stored in the Earth system: where does the energy go?" Earth System Science Data 12, Issue 3 (07 September 2020): 2013-2041. https://doi.org/10.5194/essd-12-2013-2020.

7. I. Velicogna, Yara Mohajerani, A. Geruo, F. Landerer, J. Mouginot, B. Noel, E. Rignot, T. Sutterly, M. van den Broeke, M. Wessem, D. Wiese, "Continuity of Ice Sheet Mass Loss in Greenland and Antarctica From the GRACE and GRACE Follow-On Missions." Geophysical Research Letters 47, Issue 8 (28 April 2020): e2020GL087291. https://doi.org/10.1029/2020GL087291.

8. National Snow and Ice Data Center World Glacier Monitoring Service

9. National Snow and Ice Data Center D.A. Robinson, D. K. Hall, and T. L. Mote, "MEaSUREs Northern Hemisphere Terrestrial Snow Cover Extent Daily 25km EASE-Grid 2.0, Version 1 (2017). Boulder, Colorado USA. NASA National Snow and Ice Data Center Distributed Active Archive Center. doi: https://doi.org/10.5067/MEASURES/CRYOSPHERE/nsidc-0530.001 . http://nsidc.org/cryosphere/sotc/snow_extent.html Rutgers University Global Snow Lab. Data History

10. R.S. Nerem, B.D. Beckley, J. T. Fasullo, B.D. Hamlington, D. Masters, and G.T. Mitchum, "Climate-change–driven accelerated sea-level rise detected in the altimeter era." PNAS 15, no. 9 (12 Feb. 2018): 2022-2025. https://doi.org/10.1073/pnas.1717312115.

11. https://nsidc.org/cryosphere/sotc/sea_ice.html Pan-Arctic Ice Ocean Modeling and Assimilation System (PIOMAS, Zhang and Rothrock, 2003) http://psc.apl.washington.edu/research/projects/arctic-sea-ice-volume-anomaly/ http://psc.apl.uw.edu/research/projects/projections-of-an-ice-diminished-arctic-ocean/

12. USGCRP, 2017: Climate Science Special Report: Fourth National Climate Assessment, Volume I [Wuebbles, D.J., D.W. Fahey, K.A. Hibbard, D.J. Dokken, B.C. Stewart, and T.K. Maycock (eds.)]. U.S. Global Change Research Program, Washington, DC, USA, 470 pp, https://doi.org/10.7930/j0j964j6 .

13. http://www.pmel.noaa.gov/co2/story/What+is+Ocean+Acidification%3F

14. http://www.pmel.noaa.gov/co2/story/Ocean+Acidification

15. C.L. Sabine, et al., “The Oceanic Sink for Anthropogenic CO2.” Science 305 (16 July 2004): 367-371. https://doi.org/10.1126/science.1097403.

16. Special Report on the Ocean and Cryosphere in a Changing Climate , Technical Summary, Chapter TS.5, Changing Ocean, Marine Ecosystems, and Dependent Communities, Section 5.2.2.3. https://www.ipcc.ch/srocc/chapter/technical-summary/

Header image shows clouds imitating mountains as the sun sets after midnight as seen from Denali's backcountry Unit 13 on June 14, 2019. Credit: NPS/Emily Mesner Image credit in list of evidence: Ashwin Kumar, Creative Commons Attribution-Share Alike 2.0 Generic.

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Unlocking Engagement in Middle and High School Math

A small shift in lesson presentation can encourage older students to actively participate in class.

The usual classroom symphony of settling students, chattering friends, and whispered gossip was abruptly interrupted by groans and jeers when they saw another dull math warm-up. As a new seventh-grade math teacher, I yearned to make the lessons engaging, but I felt constrained by the curriculum and struggled to stay ahead of the students, often learning the material the night before I taught it. In my focus on mastering the content, I had neglected the student experience, which, as their growing disengagement and defiance made clear, was lacking.

One day, amid the usual classroom hubbub, a sense of resignation hung heavy in the air. Students went through the motions of learning, some with glazed eyes copying notes verbatim, while others chatted animatedly about weekend plans. In stark contrast, “Sofia,” usually a beacon of participation, slumped in her chair, and “Josiah,” who often fidgeted, leaned forward intently, gesturing toward a crucial note on the whiteboard.

The usual mix of focused energy and playful banter felt off-balance, replaced by a quiet apathy. It felt like a repetitive performance, and I knew I had to offer them something more meaningful. I also knew that many of them genuinely wanted to learn, but the classroom disarray forced them into survival mode, simply hoping to avoid embarrassment or disruption.

My train of thought was interrupted by a knock at the door, barely audible over the din of student protest. My math coach entered with a smile, but my initial flicker of hope was quickly extinguished by the self-doubt gnawing at me, questioning my career choice.

A Surprising Discovery

With gentle encouragement from my coach, I launched into the lesson, my nerves on edge from a potent mix of disengaged students, my own lack of confidence, and the looming presence of an external observer. My coach had mentioned that many students indicated a desire to learn, and I held on to that thought as I wrote the first step on the board. Whether due to nerves or to sleep deprivation, I found myself pausing after each step, struggling to recall what came next. During these pauses, I noticed my students surprisingly engaged, gesturing and discussing what might happen next.

After a few instances of this, my coach pulled me aside and suggested I continue pausing and ask the students to predict the next step. The transformation was gradual at first, but it quickly spread throughout the room. Once-rowdy students were eagerly waiting, anticipating the next step. The rest of the lesson unfolded completely differently. I no longer felt burdened by the content, and the students thrived on unraveling the mystery.

Inspired by this experience, my coach and I sought to refine this concept of student engagement. I decided to use existing examples from the curriculum, sequencing them and displaying each step on a slide with animation. Students would then predict the next step. My coach expressed concern about the potential loss of momentum if no one could guess correctly. I countered with the idea of presenting two steps, like step one and step two, and asking students to compare them. We were both excited, and I eagerly began crafting the initial iterations of what I called the spark sequence .

This simple yet impactful pedagogical strategy is designed to enhance student learning and engagement while reducing teacher workload. It leverages our natural pattern recognition abilities to do the following:

• Focus student attention on key details through a step-by-step process
• Develop critical thinking by comparing similarities and differences across each step
• Boost confidence by enabling students to share their observations and build upon their understanding

New Levels of Engagement

The next day, a mix of excitement and nervousness coursed through me as I began the lesson. The usual pre-lesson grumbling commenced, but as we reached the first example, a shift was palpable. I wasn’t burdened with the explanation, and the students were once again engaged by the challenge. With two steps presented, they could freely analyze and compare. I then employed the “same-different” strategy, where students compare two things and identify similarities and differences.

While it took some students a moment to grasp the concept, the classroom chatter had transformed. Students were actively discussing their observations, their faces lighting up with understanding as the lesson progressed. A chorus of “That was fun!” confirmed their enjoyment, and they walked away from the example knowing how to solve it.

I then revisited the complete example, but instead of lecturing, I had students take turns explaining each step, collaboratively constructing a narrative.

Steps in the spark sequence

• Briefly display each step with minimal distractions.
• Think-pair-share. After each step, ask students to silently identify what’s the same and different (<10 seconds). Then, have them share their observations with a partner (15 seconds each).
• Repeat steps two and three.
• Once the sequence is complete, reveal the final outcome or justification. Allow students time to recall (10–30 seconds), then have them discuss with their partners (<1 minute).
• Wrap up: The whole class shares out. The teacher clears up any remaining misconceptions and summarizes the day’s learning.

In the end, I learned more from the students’ explanations than I had from the countless hours spent preparing the night before. In essence, I had discovered a way to engage my students, reduce my prep time, and learn alongside them. They became active participants in their education, taking the driver’s seat in their learning journey. By stepping aside and letting them take ownership, I fostered engagement and a shared learning experience for all of us.

Looking back, these moments marked a turning point. They transformed my doubts about my career choice into a deep sense of purpose. My students continue to love this strategy and the framework for discussion it provides. We’ve even incorporated it into student work, as they strive to create their own sequences or explore different applications of the strategy. It’s a place where we’ve all thrived—an oasis of learning and mutual respect, and a space where this old student has learned from his younger ones.

• Open access
• Published: 28 March 2024

High Oestrogen receptor alpha expression correlates with adverse prognosis and promotes metastasis in colorectal cancer

• Geriolda Topi 1 , 5   na1 ,
• Shakti Ranjan Satapathy 1   na1 ,
• Souvik Ghatak 1 ,
• Karin Hellman 2 ,
• Fredrik Ek 2 ,
• Roger Olsson 2 ,
• Roy Ehrnström 3 ,
• Marie-Louise Lydrup 4 &
• Anita Sjölander 1  

Cell Communication and Signaling volume  22 , Article number:  198 ( 2024 ) Cite this article

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In normal colon tissue, oestrogen receptor alpha (ERα) is expressed at low levels, while oestrogen receptor beta (ERβ) is considered the dominant subtype. However, in colon carcinomas, the ERα/β ratio is often increased, an observation that prompted us to further investigate ERα’s role in colorectal cancer (CRC). Here, we assessed ERα nuclear expression in 351 CRC patients. Among them, 119 exhibited positive ERα nuclear expression, which was significantly higher in cancer tissues than in matched normal tissues. Importantly, patients with positive nuclear ERα expression had a poor prognosis. Furthermore, positive ERα expression correlated with increased levels of the G-protein coupled cysteinyl leukotriene receptor 1 (CysLT 1 R) and nuclear β-catenin, both known tumour promoters. In mouse models, ERα expression was decreased in Cysltr1 −/− CAC (colitis-associated colon cancer) mice but increased in Apc Min/+ mice with wild-type Cysltr1 . In cell experiments, an ERα-specific agonist (PPT) increased cell survival via WNT/β-catenin signalling. ERα activation also promoted metastasis in a zebrafish xenograft model by affecting the tight junction proteins ZO-1 and Occludin. Pharmacological blockade or siRNA silencing of ERα limited cell survival and metastasis while restoring tight junction protein expression. In conclusion, these findings highlight the potential of ERα as a prognostic marker for CRC and its role in metastasis.

Introduction

Oestrogen receptor alpha (ERα), one of the two primary oestrogen receptors encoded by the ESR1 gene located on chromosome 6, plays a crucial role in the tumorigenesis of various cancers, such as breast, prostate, uterine, and ovarian cancers [ 1 , 2 , 3 , 4 ]. In normal colon mucosa, ERα expression is generally low [ 5 , 6 ]. However, studies have shown an increased ERα/β ratio in colon carcinomas [ 7 , 8 ]. Despite this discrepancy, there have been limited investigations of the prognostic significance of ERα in colorectal cancer (CRC) patients [ 9 , 10 , 11 ]. Therefore, further research is needed to elucidate the prognostic implications of ERα in CRC.

CRC is currently the third most common cancer worldwide, claiming thousands of lives annually [ 12 , 13 ]. The development of CRC is associated with genetic alterations and inflammation [ 14 , 15 ]. Mutation of the tumour suppressor gene adenomatous polyposis coli ( APC ) is present in approximately 85% of sporadic CRC cases [ 16 ]. Somatic loss of APC results in aberrant activation of the Wnt/β-catenin pathway, which is known to play a pivotal role in the development and prognosis of CRC [ 16 , 17 ]. Additionally, cysteinyl leukotrienes (CysLTs) have been implicated in colon tumorigenesis [ 18 ]. Previous studies have investigated the tumour-promoting effects of CysLT 1 R and have demonstrated its association with poor prognosis in CRC patients, as well as its tumour-promoting effects in colon cancer (CC) cell lines and several mouse models [ 19 , 20 , 21 ].

In this study, using CRC patient material, animal models, and CC cell lines, we investigated how ERα influences the prognosis of CRC patients and promotes metastasis in CRC. We report that an increase in ERα expression leads to poor outcomes in patients with CRC and that activated ERα expression stimulates the expression of tumour promoters and drives metastasis via regulation of tight junction proteins. Furthermore, we demonstrate that targeting ERα either through pharmacological inhibition or siRNA silencing could decrease the metastatic burden in preclinical models of CC.

Materials and methods

The patients involved in this study were represented from two cohorts: the Kvinno cohort, composed of a total of 333 CRC samples collected between January 1, 2008, and June 30, 2012; and the Malmö cohort, comprising 120 CRC samples collected during 1990. All the samples collected were incorporated into tissue microarrays (TMAs). Details about the study design and patient follow-up for each cohort were provided earlier [ 22 , 23 ]. Both studies were approved by the Ethics Committee at Lund University. The flowchart of patient inclusion in the study is shown in Supplementary fig. S 1 .

Tissue microarray (TMA) and immunohistochemical (IHC) analyses

The tissues from CRC samples incorporated into the TMAs were stained to evaluate the expression of ERα using a mouse anti-ERα antibody cocktail (overnight incubation, pH 6.0) and an anti-ERα monoclonal antibody D-12 (overnight incubation, pH 6.0) as mentioned earlier [ 24 ]. Additionally, anti-CysLT 1 R (overnight incubation) and anti-total β-catenin antibodies were used. Immunoreactivity was assessed by two blinded independent investigators (GT and RE) using the immunoreactive score (IRS) calculated as follows: IRS = (staining intensity) × (% of stained cells) [ 22 , 24 ]. Only the nuclear ERα expression was taken into consideration. All the cores with positive nuclear staining in more than 10% of the cells, regardless of the staining intensity, were considered positive for ERα expression. Tissue cores with less than 10% of positive nuclear staining were considered negative for ERα expression. For antibody dilutions and details, see Table  1 .

The human CC cell lines HT-29 and Caco-2 were obtained from the American Type Culture Collection (ATCC, Manassas, Virginia, USA). HT-29 cells were cultured in McCoy’s 5A medium supplemented with 10% foetal bovine serum (FBS), 1% L-glutamine and 100 μg/ml penicillin/streptomycin. Caco-2 cells were maintained in MEM supplemented with 20% foetal bovine serum (FBS), 1% L-glutamine, 1% non-essential amino acids, and 100 μg/ml penicillin/streptomycin. Both cell lines were incubated at 37 °C and 5% CO 2 . Cells were treated for 48 h with the ERα-selective agonist, PPT (40 nM) with or without treatment with the ERα-selective antagonist AZD9496 (0.3 nM) 30 min before PPT stimulation.

Quantitative real-time PCR

The isolation of total RNA from CC cells was performed using RNeasy Mini Kit (Qiagen, Hilden, Germany), and first-strand cDNA synthesis was accomplished using a cDNA synthesis kit (Invitrogen, Carlsbad, California, USA) [ 25 , 26 , 27 , 28 ]. The TaqMan probes used were specific for ESR1 (ERα, Hs01046816_m1), CYSLTR1 (CysLT 1 R, Hs00272624-s1), and CTNNB1 (β-catenin, Hs00991818_ml). The samples were analysed, expression levels were normalized to those of the endogenous housekeeping gene HPRT1 (Hs99999909_m1), and fold changes were quantified with the 2-ΔΔCt method using MxPro software (Agilent Technologies, Santa Clara, CA, USA).

Western blotting

Western blotting was performed based on a protocol described previously with minor modifications [ 25 , 26 , 27 , 28 , 29 ]. The following primary antibodies were used: anti-ERα, anti-phospho-β-catenin (Ser45/Thr41), anti-non-phospho (active)-β-catenin, and anti-total β-catenin. Anti-α-tubulin and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibodies were used as loading control antibodies as indicated in the figures. Visualization of protein expression was performed with enhanced chemiluminescence (ECL) reagents Millipore with a Bio-Rad ChemiDoc™ imaging system (Hercules, CA, USA), and densitometric analysis was performed using Bio-Rad Image Lab software. For antibody dilutions and details (see Table 1 ).

siRNA transfection in colon cancer cells

An ESR1 -specific siRNA ( siESR1 ) was employed to study ERα specificity in functional assays using CC cells in accordance with a previously reported protocol [ 27 ].

Colonosphere model

HT-29 and Caco-2 CC cell-derived colonospheres were generated as described previously [ 25 , 26 , 27 ].

Immunofluorescence analysis

Immunofluorescence analysis was performed in HT-29 and Caco-2 cells to visualize the expression of the tight junction proteins ZO-1 and Occludin as previously described [ 29 ]. Fluorescence images were captured with a Zeiss LSM 700 confocal microscope (Carl Zeiss Microscopy GmbH, Jena, Germany) using a 63× oil objective and analysed using LSM ZEN Blue software.

Immunofluorescence staining in colonospheres was performed by adopting a protocol designed for organoids with some modifications [ 30 ]. Briefly, CC cells were used to generate colonospheres as described previously [ 25 , 26 , 27 ]. After 72 h, colonospheres were harvested and cultured in Matrigel in eight-well glass-bottom chamber slides (Ibidi, GmbH, Germany). After 24 h, the medium was removed from each chamber, and the colonosphere embedded in the Matrigel was washed with 1x PBS and fixed with 4% PFA for 1 h at room temperature (RT). After fixation, the colonospheres were washed with 1x PBS and permeabilized with 0.3% Triton X-100 for 15 min at RT. Blocking was performed with 5% BSA prepared in PBS containing 0.1% Tween 20 overnight at 4 °C. Colonospheres were incubated with the primary antibody (see Table 1 for dilutions) overnight at 4 °C prior to washing with PBS containing 0.1% Tween 20. Next, incubation with a secondary antibody combined with a nuclear stain was performed for 4 h at RT prior to washing with 1x PBS containing 0.1% Tween 20. Colonospheres were mounted using Dako mounting medium (Dako, Denmark), and images were acquired with a confocal microscope (Zeiss, Germany).

Colon cancer mouse models

We evaluated the expression of ERα in two different CC mouse models. C57BL/6 J- Apc Min/+ mice (The Jackson Laboratory, Bar Harbor, Maine, USA) and in a CAC model with Cysltr1 gene disruption on the C57BL/6 N background [ 31 ]. The experimental design for the CAC model is described in detail earlier [ 32 ]. Tissues from these mice were stained with an anti-ERα antibody (1D5 + 6F11, 1:50) to evaluate ERα expression. Five mice per genotype were included, and four different sections per mouse were evaluated.

Zebrafish xenograft model

The role of ERα expression in CC cell metastasis was validated using the zebrafish xenograft model [ 25 , 26 , 27 , 33 ]. HT-29 CC cells were treated with PPT with or without AZD9496 for 48 h, labelled with Vybrant DiI and microinjected into the perivitelline space of the 2 days post-fertilization zebrafish embryos. siCTRL- or siESR1 -transfected cells were treated with PPT for 48 h before DiI labelling and were injected into the embryos. After 48 h, the metastatic spread of CC cells into the tail veins of the embryos was monitored and photographed using a Nikon fluorescence microscope (Nikon Eclipse 80i, USA). The mean fluorescence intensity (MFI) of the metastatic cells was measured using ImageJ software (NIH, USA). The zebrafish housing and maintenance were as described before [ 34 ].

Statistical analysis

Continuous variables were compared using the Mann-Whitney U test or Student’s t test as indicated, and categorical variables were compared using the chi-square test. Kaplan–Meier- survival curves were compared with the log-rank test. Statistical analyses were performed using SPSS version 23.0 (SPSS, IBM, Armonk, NY, USA) and GraphPad Prism version 9 (GraphPad Software, Inc., San Diego, CA, USA). All tests were two sided, and P values < 0.05 were considered to indicate a statistically significant difference.

Public CRC dataset analysis

We also used public CRC patient data. The mRNA expression based microarray data from the GSE39582 dataset were downloaded. ERα gene expression in this public database was categorised into high ERα and low ERα. Data for a total of 566 patients were included in the study. The mRNA expression data in the dataset were normalized using the TMM method and were log2 transformed for further analysis.

ERα expression and its prognostic association in colorectal cancer patients

Recently, we reported that KRAS oncogenic mutation correlates with positive ERα expression in CC patients [ 24 ]. However, the involvement of ERα in CC growth and metastasis is not yet clear. Here, using paired normal and tumour samples of CRC patients, we found consistently elevated expression of ERα in the tumour tissue compared to the normal mucosal tissue (Fig.  1 A). Previously, we reported that high ERα expression in CRC patients is associated with poor overall survival (OS) [ 24 ]. Here, we combined the same cohort (Kvinno cohort, n  = 270) with another cohort of patients with CRC (Malmö cohort, n  = 67) and grouped the patients based on ERα expression (positive or negative) (Supplementary Fig. S 1 ). After excluding patients from both cohorts with metastatic disease or an unknown recurrence status or date, 233 patients were available for disease free survival (DFS) analysis. The median follow-up times for the Kvinno cohort were 5.8 years for OS and 5 years for DFS. For the Malmö cohort, the median follow-up time for OS was 10 years. To investigate the prognostic role of ERα expression in CRC patients, we performed Cox regression analysis and evaluated OS and DFS. Patients with negative ERα expression had a significantly reduced risk for overall mortality of 61% (HR, 0.39; CI, 0.28–0.55) after adjustment for age and TNM stage (Fig. 1 B). This result was consistent in the subgroups of patients with stage I-III CRC disease and colon cancer and, interestingly, also in the subgroup of patients with rectal cancer, in which negative ERα expression showed the greatest benefit, with a reduction in overall mortality of 80% (Fig. 1 B-D). Similar results were obtained for patients who did not receive any adjuvant treatment after CRC surgery, for both patients with colon or rectal cancer (Fig. 1 E-G). Together these data indicates that high ERα expression is associated with poor prognosis in CRC patients.

ERα expression and its prognostic association in colorectal cancer (CRC) patients. A Western blot of ERα protein expression in six paired colon cancer (CC) patient samples of normal (N) and tumour (T) colon tissue. GAPDH was used as a loading control. Graph showing the densitometric analysis of ERα protein expression compared between normal (N) and tumour (T) tissues. Kaplan-Meier survival curves for OS: B  multivariate model for CRC patients with stage I-III disease, n  = 299; C  multivariate model for patients with colon cancer, n  = 271; and D  multivariate model for patients with rectal cancer, n  = 63. Kaplan-Meier survival curves for DFS: E  multivariate model for patients who did not receive adjuvant treatment after surgery, n  = 128; F  multivariate model for patients with colon cancer, n  = 142; and G  multivariate model for patients with rectal cancer, n  = 41. P values < 0.5 were considered significant and determined by the log-rank test

ERα expression positively correlates with tumour promoter expression in colon cancer

In this cohort, we previously showed that the poor prognosis of the included patients was associated with high CysLT 1 R and nuclear β-catenin expression. These two tumour promoters in colon cancer are known to act through a CysLT 1 R/Wnt-β-catenin signalling axis [ 25 , 28 , 32 , 35 ]. Here, we tested for possible associations between ERα and CysLT 1 R and/or β-catenin. As expected, we noted higher expression of CysLT 1 R and nuclear β-catenin in patients with positive ERα expression ( n  = 185) compared to patients with negative ERα expression ( n  = 82) (Fig.  2 A, B). This finding was further validated by analysis of a public CRC dataset (GSE39582, n  = 566), which revealed a significant positive correlation between the mRNA level of CYSLTR1 or CTNNB1 (β-catenin) and ESR1 (ERα) (Fig. 2 C, D). In addition, patients with high ESR1 expression showed higher transcript levels of both CYSLTR1 and CTNNB1 (Fig. 2 E, F).

ERα expression positively correlates with tumour promoter expression in colon cancer. Mean immuno-reactive score (IRS) for ( A ) Cysteinyl leukotriene receptor 1 (CysLT 1 R) and ( B ) nuclear β-catenin expression in colorectal cancer (CRC) patients ( n  = 267) with negative and positive ERα expression. XY scatter plots of the mRNA levels of ( C ) ERα ( ESR1 ) and CysLT 1 R ( CYSLTR1 ) and ( D ) ERα ( ESR1 ) and β-catenin ( CTNNB1 ) in the GSE39582 public dataset ( n  = 566) of CRC patients. Violin plots showing the mRNA expression of ( E ) CYSLTR1 and ( F ) CTNNB1 in CRC patients with low or high ESR1 expression. G Immunohistochemical (IHC) images of ERα expression in the colons of wild-type ( WT ) and Cysltr1 knockout ( Cysltr1 −/− ) mice in a colitis-associated colon cancer (CAC) mouse model (n = 5). Bar graph showing the IRS of ERα expression compared between the WT and Cysltr1 −/− mouse groups. H IHC images of ERα expression in the colons of WT and Apc Min/+ mice, n  = 5. Bar graph showing the IRS of ERα expression in the WT and Cysltr1 −/− mouse groups. For both mouse models, four random regions of interest (ROIs; marked with dotted lines) in colon tissue were evaluated for each mouse. Representative images of one ROI are shown as insets. The scale bars represent 2 mm ( G ) and 500 μm ( H ) in the image of the whole colon and 50 μm in the zoomed insets. P values were calculated using an unpaired Student’s t test for the bar graphs in G and H. Relative mRNA expression levels of ESR1 , CYSLTR1 , and CTNNB1 in ( I ) HT-29 and ( J ) Caco-2 CC cells after treatment with PPT (ERα specific agonist, 40 nM) or AZD9496 (ERα specific antagonist, 0.3 nM for 30 min) alone or in combination of PPT (40 nM) with AZD9496 (0.3 nM for 30 min before the PPT treatment). The data are presented as the mean ± SEM ( n  = 3 independent experiments). P values < 0.5 were considered significant were calculated using an unpaired Student’s t test

Next, we used both the colitis-associated colon cancer (CAC) mouse model and spontaneous CC mouse model ( Apc Min/+ ) with either the functional presence or functional absence of Cysltr1 (n = five mice per genotype) (Fig. 2 G, H). Cysltr1 −/− mice are reported to exhibit a less aggressive tumour phenotype, with fewer polyps/tumours in their colon [ 32 , 35 ]. Colon tissue sections from mice lacking functional Cysltr1 ( Cysltr1 −/− ) showed two-fold reduced ERα expression compared to those from WT mice (Fig. 2 G). On the other hand, we found that Apc Min/+ mice, which have higher amount of β-catenin accumulation in the nucleus due to the Apc mutation and are reported to develop more and larger intestinal polyps [ 36 ], had >two-fold higher nuclear ERα expression than mice with wild-type Apc (Fig. 2 H).

Furthermore, we evaluated the basal mRNA levels of ERα expression in different CC cell lines, using the MCF7 breast cancer cell line, with very high expression of ERα, as a positive control (data not shown). Based on these results we used the ERα expressing cell lines HT-29 and Caco-2 cells for further studies. We pharmacologically induced or blocked ERα expression in HT-29 and Caco-2 cells by treatment with the ERα-selective agonist PPT alone or ERα-specific antagonist AZD9496 or combination of both (AZD9496 + PPT) (see Materials and Methods for experimental details). We observed a more than 30–50% increase in the mRNA expression levels of both CYSLTR1 and CTNNB1 with PPT treatment, and this increase was restricted after AZD9496 alone treatment or in PPT stimulated cells pre-treated with AZD9496 (PPT + AZD9496) in both HT-29 and Caco-2 cells (Fig. 2 I, J). No statistical significance was noted between AZD9496 or AZD9496 + PPT experimental groups (Fig. 2 I, J). Taken together, these results indicate a direct involvement of ERα in promoting the expression of the tumorigenic markers CysLT 1 R and β-catenin in CC.

ERα activation in colon cancer cells promotes survival

WNT/β-catenin signalling in CC is known to support tumour growth via nuclear β-catenin. Based on the observations in Fig. 2 B and H, we next sought to investigate whether activation of ERα can promote cell survival via the WNT/β-catenin pathway. Indeed, we observed a 2-fold increase in the number of colonies formed by both HT-29 and Caco-2 CC cells treated with PPT compared to untreated control cells, and the number of colonies formed by cells pretreated with AZD9496 was substantially reduced (Fig.  3 A). Furthermore, western blot analysis of HT-29 and Caco-2 cells stimulated with PPT showed increased levels of non-phosphorylated active β-catenin compared to unstimulated cells (Fig. 3 B). Interestingly, the increase in the β-catenin level was inhibited in cells pretreated with AZD9496 prior to PPT stimulation, possibly due to the significant increase in phosphorylated β-catenin (three-fold for HT-29 and six-fold for Caco-2 cells), which is known to be rapidly degraded by ubiquitination.

ERα activation in colon cancer cells promotes survival. A Alterations in the colonies formed by HT-29 and Caco-2 colon cancer (CC) cells treated with PPT (40 nM) alone for 48 h or in combination with AZD9496 (0.3 nM, for 30 min before PPT treatment). Bar graphs show the percentage of survival and are representative of n  = 3 independent experiments. B Western blots showing the protein levels of phospho-β-catenin (Ser33/37/Thr41), non-phospho (active)-β-catenin, total β-catenin, and ERα in HT-29 and Caco-2 cells untreated or treated with PPT (40 nM) alone or in combination with AZD9496 (0.3 nM, for 30 min). Graphs showing the densitometric analysis of alterations in phospho- and non-phospho (active)-β-catenin and ERα protein levels as percentages of the loading control (α-tubulin). The blots are representative of n = 3 independent experiments. C Alterations in the colonies formed by HT-29 and Caco-2 cells transfected with either siCTRL or siESR1 prior to PPT (40 nM) treatment for 48 h. The graphs show the percentage of survival in each group. D Western blots showing the protein levels of non-phospho (active)-β-catenin, total β-catenin, and ERα in both HT-29 and Caco-2 cells transfected with either siCTRL or siESR1 prior to PPT (40 nM) treatment. Graphs showing the densitometric analysis of alterations in ERα and non-phospho (active)-β-catenin protein levels as percentages of the loading control (α-tubulin). The blots are representative of n = 3 independent experiments. The data are presented as the means ± SEMs. P values < 0.5 were considered significant calculated using an unpaired Student’s t test

To further validate the involvement of ERα in CC cell survival, we employed ESR1 siRNA to silence functional ERα, and the results were compared with those in siCTRL -transfected cells (Fig. 3 C, D). Interestingly, we noted a significantly reduced colony number in the siESR1 -transfected cell group compared to the siCTRL -transfected cell group (Fig. 3 C). Treatment with PPT did not significantly affect the colony number in the siESR1 -transfected cell group. Moreover, the whole-cell lysates of HT-29 and Caco-2 cells transfected with siESR1 showed a reduction in the level of active β-catenin, supporting the earlier observation of the increase in active β-catenin in PPT-treated CC cells (Fig. 3 D).

ERα promotes colon cancer cell metastasis by downregulating tight junction proteins

As reported earlier, ERβ activation could reduce metastasis in CC, with a reduced number of migrated cells observed in vitro and in vivo [ 27 ] . Here, we intended to explore the metastatic potential of ERα, which is known to act in an opposite manner in breast cancer [ 37 ]. We used a public dataset of CC patients with liver metastasis ( n  = 18) and found a strong and significant positive correlation between CYSLTR1 and ESR1 (R = 0.77, p  < 0.005) as well as between CTNNB1 and ESR1 expression (R = 0.62, p  < 0.041) (Fig.  4 A, B). Next, we tested the impact of ERα activation on the migration capacity of CC cells in vitro using wound-healing assay (Supplementary Fig. S 2 A). HT-29 cells stimulated with PPT showed more than 2-fold increase in the wound closure compared to unstimulated control. Interestingly, cells pre-treated with AZD9496 showed reduced percentage (5-fold) of wound closure compared to the PPT treated cells suggesting an involvement of ERα in the migration of CC cells. Next, to validate this observation in vivo, we used a transgenic zebrafish xenograft model established with HT-29 cells, interestingly, we observed a higher number of embryos with metastasis as well as an increased metastatic burden in the tail veins of the embryos in the group injected with PPT-treated HT-29 cells (M1 = 24; M0 = 6) compared to the group injected with untreated HT-29 cells (M1 = 10; M0 = 20) (Fig. 4 C, D). The increase in metastasis was inhibited in the embryos injected with AZD9496 + PPT-treated HT-29 cells (M1 = 5; M0 = 38). PPT treatment not only resulted in a greater number of embryos with metastasis but also was able to establish a high metastatic burden, as evidenced by quantification of the mean fluorescence intensity (MFI) (mean MFI in the CTRL group = 187; PPT group = 330; AZD9496 + PPT group = 172) (Fig. 4 D’).

Activation of ERα promotes colon cancer cell metastasis. An external dataset composed of data for CC patients with liver metastasis (GSE77955, n  = 18) was used to analyse the correlations of CysLT 1 R ( CYSLTR1 ) and β-catenin ( CTNNB1 ) with ERα ( ESR1 ). The scatter plots show the positive correlations between ESR1 and both ( A ) CYSLTR1 and ( B ) CTNNB1 . C Schematic cartoon showing the zebrafish embryo-based colon cancer metastasis model. DiI-labelled HT-29 cells left untreated or treated with PPT alone or in combination with AZD9496 were injected into the perivitelline space of 2 dpf zebrafish embryos, and the embryos were incubated for 48 h. Images showing the metastatic spread of HT-29 cells in the tail veins of zebrafish embryos in each group (CTRL, n  = 30; PPT, n = 30; AZD9496 + PPT, n  = 43). Scale bars: full-size images; 10 μm, insets; 2 μm. The insets show the regions enclosed in the dotted lines in the full-size tail images. The arrows point to the metastatic foci and transendothelial migration of cancer cells. D Graphs showing the number of embryos with (M1, mets) or without (M0, no mets) metastasis in each group and D′, quantification of tail vein metastasis using the mean fluorescence intensity (MFI) of the embryos with metastasis. E Western blots showing the expression of the tight junction protein ZO-1 in HT-29 and Caco-2 cells treated with PPT (40 nM) alone or in combination with AZD9496 (0.3 nM, 30 min). Graph showing the densitometric analysis of alterations in protein expression as a percentage of the loading control (α-tubulin). The blots are representative of n = 3 independent experiments. For the bar graphs, unpaired t-test was used. F Immunofluorescence analysis of ZO-1 and Occludin expression in HT-29 cells treated with PPT (40 nM) alone or in combination with AZD9496 (0.3 nM, 30 min). Greyscale images (insets) showing a representative region of interest (dotted line) for ZO-1 and Occludin staining. Scale bars: full-size images; 5 μm, insets; 1 μm. Violin plots showing the mean fluorescence intensity of ZO-1 (CTRL, n  = 116; PPT, n  = 105, AZD9496 + PPT, n  = 107) and Occludin in random cell-cell junctions (CTRL, n  = 103; PPT, n  = 110, AZD9496 + PPT, n  = 108). P values were calculated with unpaired Student’s t test. The arrows indicate gaps in ZO-1 expression. G Immunofluorescence analysis of ZO-1 and Occludin expression in Caco-2 cells treated with PPT (40 nM) alone or in combination with AZD9496 (0.3 nM, 30 min). Greyscale images (insets) showing representative regions of interest for ZO-1 and Occludin staining. Scale bars: full-size images; 5 μm, insets; 1 μm. Violin plots showing the mean fluorescence intensity of ZO-1 (CTRL, n  = 108; PPT, n  = 116, AZD9496 + PPT, n  = 105) and Occludin (CTRL, n = 105; PPT, n  = 115, AZD9496 + PPT, n = 116) in random cell-cell junctions. The arrows indicate gaps in ZO-1 or Occludin expression. H Immunofluorescence analysis of ZO-1 in HT-29 cell-derived colonospheres treated with PPT (40 nM) alone or in combination with AZD9496 (0.3 nM, 30 min). Scale bars: 10 μm. Violin plot showing the mean fluorescence intensity of ZO-1 in random colonospheres (CTRL, n  = 31; PPT, n  = 28, AZD9496 + PPT, n  = 30). The arrows indicate ZO-1 expression in the disseminated cells from the colonospheres. I Immunofluorescence analysis of ZO-1 in Caco-2 CC cell-derived colonospheres treated with PPT (40 nM) alone or in combination with AZD9496 (0.3 nM, 30 min). Scale bars: 10 μm. Violin plot showing the mean fluorescence intensity of ZO-1 in random colonospheres (CTRL, n  = 29; PPT, n  = 26, AZD9496 + PPT, n = 28). The data are presented as the mean ± SEM of three experiments. P values < 0.5 were considered significant were calculated using the chi-square test in D and an unpaired Student’s t test in D′-I

Cancer cell dissemination due to loss of cell-cell junctions is a key and early step in metastasis. Tight junction proteins are responsible for maintaining physical connections between epithelial cells. Important among tight junction proteins is the ZO-1/Occludin complex [ 38 ]. Therefore, we next sought to determine whether ERα-mediated metastasis in CC occurs via disruption of tight junctions with a possible change of the ZO-1 complex. Indeed, we found a decrease in ZO-1 expression in HT-29 (50%) and Caco-2 (40%) cells treated with PPT (Fig. 4 E). This was further supported by immunofluorescence staining of HT-29 and Caco-2 cells, where it was clearly evident that there was disruption of ZO-1 expression in cells after PPT treatment, while in both untreated and AZD9496-treated cells, ZO-1 expression remained unchanged (Fig. 4 F, G). Furthermore, Occludin, a member of the ZO-1 complex, was expressed at significantly lower levels in PPT-treated HT-29 or Caco-2 cells owing to disrupted cell-cell junctions (Fig. 4 F, G). Moreover, analysis of ZO-1 expression in HT-29 and Caco-2 cell-derived colonospheres validated the above observations (Fig. 4 H, I). These results indicate the metastasis-promoting role of ERα induction in CC cells and the beneficial effects of antagonizing ERα expression by treatment with a selective antagonist such as AZD9496.

The functional absence of ERα inhibits colon cancer cell metastasis

Next, we tested the effect of the functional absence of ESR1 on colon cancer metastasis in vitro as well as in vivo. Wound healing assay performed in the siCTRL or siESR1 transfected cells with or without PPT treatment showed > 2-fold increase in wound closure in the siCTRL group treated with PPT (Supplementary fig. S 2 B), while siESR1 transfected cells failed to show significant wound closure even after PPT treatment.

While injection of PPT-treated siCTRL -transfected cells resulted in metastasis in a higher number of embryos (embryos with metastasis (M1) = 3; embryos with no metastasis (M0) = 29), siESR1 transfection failed to result in metastasis even after PPT treatment (M1 = 5, M0 = 24) (Fig.  5 A, A’). Zebrafish embryos injected with HT-29 cells transfected with siESR1 showed reduced tail vein metastasis compared to embryos injected with siCTRL -transfected HT-29 cells, with a 3-fold reduction in the MFI indicating the metastatic burden in embryos with metastasis (Fig. 5 A’).

Functional absence of ERα inhibits colon cancer cell metastasis. DiI-labelled HT-29 cells transfected with either siCTRL or siESR1 and treated with or without PPT for 48 h were injected into the perivitelline space of 2 dpf zebrafish embryos, and the embryos were incubated for 48 h. A Images showing the metastatic spread of HT-29 cells in the tail veins of zebrafish embryos in each group ( siCTRL ; CTRL, n  = 30; PPT, n = 30; siESR1 ; CTRL, n  = 33, PPT, n  = 32). Graphs showing A ’, the number of embryos with (M1, mets) or without metastasis (M0, no mets) in each group and A ”, the quantification of tail vein metastasis using the mean fluorescence intensity (MFI) of the embryos with metastasis (M1 group). Scale bars: full-size images; 10 μm, insets; 2 μm. The insets show the regions enclosed in the dotted lines in the full-size tail images. The arrows point to the metastatic foci and transendothelial migration of cancer cells. B Western blots showing the expression of the tight junction protein ZO-1 in HT-29 and Caco-2 cells transfected with either siCTRL or siESR1 prior to PPT (40 nM) treatment. Graphs showing the densitometric analysis of alterations in protein expression as a percentage of the loading control (α-tubulin). The blots are representative of n = 3 independent experiments. C Immunofluorescence analysis of ZO-1 and Occludin expression in HT-29 cells transfected with either siCTRL or siESR1 prior to treatment with the ERα agonist PPT (40 nM). Greyscale images (insets) showing representative regions of interest for ZO-1 and Occludin staining. Scale bars: full-size images; 5 μm, insets; 1 μm. Violin plots showing the mean fluorescence intensity of ZO-1 ( siCTRL (CTRL, n  = 105; PPT, n = 115), siESR1 (CTRL, n  = 108; PPT, n = 116)) and Occludin ( siCTRL (CTRL, n = 105; PPT, n = 105), siESR1 (CTRL, n  = 107; PPT, n  = 102)) in random cell-cell junctions. The arrows indicate gaps in ZO-1 or Occludin expression. D Immunofluorescence analysis of ZO-1 and Occludin expression in Caco-2 cells transfected with either siCTRL or siESR1 prior to treatment with the ERα agonist PPT (40 nM). Greyscale images (insets) showing representative regions of interest for ZO-1 and Occludin staining. Scale bars: full-size images; 5 μm, insets; 1 μm. Violin plots showing the mean fluorescence intensity of ZO-1 and Occludin in random cell junctions. For ZO-1 staining in the siCTRL -transfected group (CTRL, n = 105; PPT, n = 105) and in the siESR1 -transfected group (CTRL, n = 108; PPT, n = 105), random cell junctions were evaluated. For Occludin staining in the siCTRL -transfected group (CTRL, n = 105; PPT, n = 105) and in the siESR1 -transfected group (CTRL, n = 102; PPT, n = 108), random cell junctions were evaluated. The arrows indicate gaps in ZO-1 or Occludin expression. Immunofluorescence analysis of ZO-1 in colonospheres derived from either siCTRL or siESR1 transfected ( E ) HT-29 and ( F ) Caco-2 cells. Scale bars: 10 μm. Violin plots showing the mean fluorescence intensity of ZO-1 in random ( E’ ) HT-29 ( siCTRL , n = 30; siESR1 , n = 32) or ( F′ ) Caco-2 ( siCTRL , n  = 28; siESR1 , n  = 31) colonospheres. The MFIs of the indicated proteins were measured using ImageJ software (NIH, USA). G Graphical representation of the summary of the study. Upon binding to the agonist PPT, ERα dimerizes and shuttles into the nucleus. This upregulates the transcription of CYSLTR1 and CTNNB1 . In addition, it promotes metastasis by disrupting the tight junction proteins ZO-1 and Occludin. However, blocking the binding of PPT to ERα by employing an antagonist, AZD9496, prevents the activation and hence the dimerization of the receptor. This further leads to downregulation of CYSLTR1 and CTNNB1 and upregulation of the tight junction proteins ZO-1 and Occludin. The data are presented as the mean ± SEM of three experiments. P values were calculated with the chi-square test for A’ and unpaired Student’s t test for A”, B-F

We observed a reduced number of embryos with tail metastasis upon ESR1 silencing ( siESR1 ) (M1, n  = 5) (Fig. 5 A), we posited that cells, with ESR1 gene silencing, might be tightly adhered to each other, indicating increased ZO-1 expression. Initially, we employed western blotting to assess ZO-1 expression under the mentioned experimental conditions. As observed in Fig. 4 ; in Fig. 5 B, we noted reduced ZO-1 expression in whole-cell lysates of siCTRL -transfected HT-29 (50%) and Caco-2 (50%) cells after PPT treatment. Surprisingly, we found a similar expression in the siESR1 -transfected control group compared to the siCTRL -transfected control group in HT-29 cells but not in Caco-2 cells (Fig. 5 B). However, ZO-1 expression in siESR1 -transfected HT-29 and Caco-2 CC cells remained unaffected even after PPT treatment (Fig. 5 B). To validate this, we performed immunofluorescence analysis in HT-29 and Caco-2 CC cells for visual confirmation. Indeed, in the immunofluorescence images, elevated ZO-1 expression was evident at the ‘cell-cell junctions’ of siESR1 transfected HT-29 and Caco-2 cells (Fig. 5 C, D). Finally, HT-29 and Caco-2 CC cell-derived colonospheres lacking functional ESR1 (derived from siESR1- transfected cells) showed intact ZO-1 expression compared to their siCTRL -transfected counterparts (Fig. 5 E, E’, F, F′). Taken together, these observations indicate a role of ERα in promoting and supporting cell survival and metastasis in CC (Fig. 5 G).

In this study, we identified the cellular mechanisms by which the nuclear receptor ERα drives metastasis and confers a worse prognosis of CRC patients (Fig. 1 ). The expression of ERα positively correlates with that of tumour promoters in CC. Activated ERα increases the level of active β-catenin (Figs.  2  and  3 ) and disrupts the tight junction proteins ZO-1 and Occludin (Figs.  4  and  5 ), leading to increased CC metastasis (Figs. 4  and  5 ).

Oestrogen receptors are well studied in breast and cervical cancers [ 2 , 3 , 4 ]. In colon cancer, ERβ plays an important role in limiting tumour progression and metastasis [ 5 , 39 , 40 ]. We have also previously reported that activating ERβ with the selective agonist ERB-041 reduced CC cell survival, induced apoptosis, and inhibited metastatic spread [ 27 ]. However, few studies have explored ERα expression levels in CC tissues. In our earlier report [ 24 ], we showed the prognostic relevance of ERα expression in CRC patients with higher ERα expression in tumour tissue than in normal mucosa. In addition, we established the importance of considering both ERα and ERβ protein expression for better predicting the prognosis of CRC patients [ 24 ].

Here, we investigated the correlation of ERα expression with that of tumour promoters in CRC. We noted a poorer prognosis in CRC patients with high ERα expression compared to the patients with low ERα expression (Fig. 1 ), which could be due to the significant correlations between ERα and both CysLT 1 R and β-catenin, all known tumour promoters [ 18 , 20 , 25 , 28 , 32 , 35 ]. Our results were supported by mRNA expression data from public databases (Fig. 2 C, D), which showed positive correlations between ESR1 and both CYSLTR1 and CTNNB1 mRNA levels in CRC patients. Moreover, we have reported earlier that CysLT 1 R, and β-catenin expression are positively correlated in CRC patients and patients with high CysLT 1 R, and high nuclear β-catenin have poorer prognosis [ 22 ]. To strengthen our findings, we examined the expression of ERα in two different CC mouse models. Cysltr1 −/− mice, known to have a less aggressive phenotype, exhibited decreased ERα expression, while Apc Min/+ mice showed higher ERα expression (Fig. 2 G, H). The Apc Min/+ mouse model exhibits activation of Wnt/β-catenin signalling, which promotes the translocation of β-catenin into the nucleus and positively correlates with CysLT 1 R expression as per our earlier report [ 32 ]. In addition, using both colon and breast cancer cells, Kouzmenko et al. have shown interaction between ERα and β-catenin via immunoprecipitation [ 41 ]. Taken together this indicate the possibility of an ERα/CysLT 1 R/Wnt signalling axis.

To maintain normal epithelial tissue integrity and cell polarity, tight junctions sustain cell-cell adhesion and regulates intercellular signalling [ 42 ]. Loss of tight junctions in a cluster of cancer cells in the primary tumour could result in dissemination and metastasis [ 42 ]. ZO-1 is a member of the tight-junction family of proteins (including Occludin) expressed in epithelial and endothelial cells and positively associated with β-catenin expression [ 43 , 44 ]. Similar to ZO-1, loss of membrane β-catenin is associated with the migratory phenotype of cancer cells. Previously, the expression levels of the tight junction proteins ZO-1 and Occludin were reported to be reduced after oestrogen treatment, resulting in ERβ activation in human gut tissue [ 45 ]. Zhou et al. used both male and female gut tissues to highlight that ZO-1 is crucial for maintaining epithelial integrity in human gut tissue. Moreover, ZO-1 loss increases gut permeability and confers vulnerability to mucosal pathogens [ 45 ]. In a more recent study, Li et al. emphasized the metastasis-promoting role of ZO-1 in KRAS mut CRC patients in a sex-specific manner [ 46 ].

We noted that pharmacological induction of ERα expression with a specific agonist, PPT, reduced the protein levels of ZO-1 and Occludin, which were restored using the specific antagonist AZD9496 (Fig. 4 F, G). This is in coordination with the alteration noted in the β-catenin expression in the mRNA or protein level (Figs. 2 I, J; 3 C). Silencing of ESR1 (by siESR1 transfection) in CC cells also protected ZO-1 and Occludin expression. This was also reflected in the number of embryos with metastasis in the zebrafish model (Fig. 4 C). Compared to injection of either AZD9496-treated cells or siESR1 -transfected cells, injection of CC cells with reduced expression of tight junction proteins upon PPT treatment resulted in an increased number of embryos with tail vein metastasis (Figs. 4 C and  5 A). This indicates that tight junction disruption is a prerequisite step in ERα-mediated CC metastasis.

Taken together, our findings show that positive expression of ERα correlates with poor prognosis and metastasis in CRC patients. These findings highlight the potential therapeutic opportunities of blocking ERα expression in CRC [ 47 ]. However, further mechanistic studies are needed to analyse the pathway in more detail. The mechanistic insights provided here could be used as a framework for the development of precision therapeutic agents targeting ERα in metastatic CRC, particularly in cases where its expression is increased, such as cases in female patients with Lynch syndrome and breast or colorectal cancer.

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Acknowledgements

We would like to acknowledge Kristina Ekstöm-Holka from the Tissue Micro-Array Centre at Lund University, Malmö, Sweden, for her assistance with immunohistochemistry analysis.

Open access funding provided by Lund University. This study was supported by grants to AS from the Malmö University Hospital Cancer Foundation, the Swedish Cancer Foundation Grant (21 1453), and the Governmental Funding of Clinical Research within the National Health Services and to SRS from the Royal Physiographic Society in Lund (Per Eric and Ulla Schyberg research grant) and the Crafoord Foundation. The funding agencies had no influence on the study design, data collection, data analysis, data interpretation or report writing.

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Geriolda Topi and Shakti Ranjan Satapathy contributed equally to this work.

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Division of Cell and Experimental Pathology, Department of Translational Medicine, Lund University, Malmö, Sweden

Geriolda Topi, Shakti Ranjan Satapathy, Souvik Ghatak & Anita Sjölander

Chemical Biology & Therapeutics Group, Department of Experimental Medical Science, Lund University, Lund, Sweden

Karin Hellman, Fredrik Ek & Roger Olsson

Department of Pathology, Skåne University Hospital, Malmö, Sweden

Roy Ehrnström

Division of Surgery, Skåne University Hospital, Malmö, Sweden

Marie-Louise Lydrup

Department of Endocrinology, Skåne University Hospital, Malmö, Sweden

Geriolda Topi

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Conception and design: GT, SRS, and AS. Development of methodology: GT, SRS, and SG. Acquisition of data: GT, SRS, SG, KH, FE, and RO. Analysis and interpretation of data: GT, SRS, and SG. Writing, review, and/or revision of the manuscript: SRS, GT, and AS. Study supervision: SRS and AS. All authors have read and approved the final version of the manuscript.

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Topi, G., Satapathy, S.R., Ghatak, S. et al. High Oestrogen receptor alpha expression correlates with adverse prognosis and promotes metastasis in colorectal cancer. Cell Commun Signal 22 , 198 (2024). https://doi.org/10.1186/s12964-024-01582-1

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Received : 12 October 2023

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DOI : https://doi.org/10.1186/s12964-024-01582-1

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