Egr1 and Egr2 regulate opposite transcriptional programs in macrophages

Project Information

MacrophageEGR

Grant agreement ID: 895411

DOI

10.3030/895411

Project closed

EC signature date 10 March 2020

Start date 1 September 2021

End date 31 August 2023

Funded under

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Total cost

€ 175 572,48

EU contribution

€ 175 572,48

Coordinated by

STICHTING AMSTERDAM UMC
Netherlands

Periodic Reporting for period 1 - MacrophageEGR (Egr1 and Egr2 regulate opposite transcriptional programs in macrophages)

Reporting period: 2021-09-01 to 2023-08-31

Summary of the context and overall objectives of the project

My research aims to understand why two similar proteins, EGR1 and EGR2, have different effects on the activation of genes related to inflammation. Specifically, it investigates the molecular mechanisms behind these differences in gene activation.

Understanding how these proteins regulate inflammation-related genes can have significant implications for the development of therapies for various diseases associated with inflammation, such as autoimmune disorders, infections, and cardiovascular diseases. By gaining insights into the molecular processes, researchers can potentially identify new targets for drug development and improve our understanding of the immune system.

Objectives:

1: Investigate the differences between EGR1 and EGR2 in their ability to activate genes related to inflammation. This includes studying the timing of their activity and their interactions with other proteins in different strains of cells.

2: Examine the effects of removing or reducing EGR1 and EGR2 in macrophages (a type of immune cell) on gene activation and cell behavior. This helps confirm the findings from Objective 1 and provides insights into how these proteins influence cell phenotype.

3: Study the role of EGR2 in real-life disease models, specifically in the context of parasitic infections and atherosclerosis. This objective aims to understand how EGR2 affects disease outcomes in living organisms, providing valuable insights into its potential as a therapeutic target.

Overall, the research seeks to uncover the molecular mechanisms behind the behavior of EGR1 and EGR2 in controlling inflammation-related genes and their significance in disease contexts.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

Overview of Results:

In the first objective, I conducted experiments to understand why two proteins, EGR1 and EGR2, which have similar DNA binding characteristics, have different effects on genes related to inflammation. I performed experiments using a technique called ChIP-seq in macrophages from five different strains of mice. This involved studying when these proteins are active in response to different stimuli. I also analyzed the genetic motifs that influence the binding of these proteins to DNA.

In the second objective, I investigated the effects of removing or reducing EGR1 and EGR2 in macrophages. I confirmed that EGR2 plays a role in enhancing the activation of certain genes related to inflammation after a specific stimulus (IL-4). I used genetic techniques to delete EGR2 in mouse cells and performed RNA-seq experiments to understand how it affects gene expression. I also studied these proteins in human macrophages.

In the third objective, I studied the role of EGR2 in live disease models. I chose to focus on a parasitic infection and atherosclerosis (plaque buildup in arteries). I found that EGR2 had a minor impact on the outcomes of these diseases, such as the number of parasites or the size of plaques in the arteries. However, I did observe changes in the types of immune cells present in certain disease contexts.

Exploitation and Dissemination:

The work performed has expanded the researcher's scientific knowledge and technical skills. It has also helped secure future grants and career advancements. I have shared my findings at various scientific conferences and meetings, both through oral and poster presentations. The impact on peers and society is expected to increase once the research is published in scientific journals and the data is made accessible through online databases. Additionally, I mentored and transferred knowledge to several students during the project.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

I discovered that EGR2 plays a significant role in enhancing the activation of genes related to inflammation, especially after 24 hours of IL-4 stimulation. I also identified motifs in DNA that affect the binding of EGR2 and other transcription factors, shedding light on the mechanisms behind gene regulation. Confirming the findings from Objective 1, I showed that EGR2 indeed influences the activation of genes related to inflammation. I found that EGR2 regulates a substantial portion of these genes and plays a role in cytokine production and signaling as the deletion of EGR2 led to decreased gene activation after stimulation with IL-4. In disease models, I found that EGR2 only had a minor impact on the outcomes of parasitic infection and atherosclerosis. However, I did observe changes in the types of immune cells present in the liver during infection and in the size of plaques in the arteries.

EGR1-EGR2 in macrophages