SPUR 2020: Molecular Activation of Quiescent Cardiac Fibroblast to Myofibroblast In Heart Failure Induced by Myocardial Injury

Background

Ischemic heart disease and heart failure HF is the leading cause of death and disability worldwide. Activated myofibroblast induced by injury or heart diseases adversely affects the cardiac function by depositing excess extracellular matrix and remodel the structure of myocardium resulting in increased stiffness and reduced compliance of cardiac muscle. Currently there are no effective interventions that specifically attenuate or reverse pathological cardiac fibrosis. To identify and quantify the cellular populations that correlated with quiescent and activated cardiac fibroblasts, we performed single cell RNA sequencing of non-myocytes from human non-failing hearts (donor hearts that were not allocated for transplant for non-cardiac related reasons) and failing hearts (HF). RNA expression profiles showed 2 distinct clusters characteristic of fibroblasts and myofibroblasts respectively in HF whereas non-failing heart donor reveal only one cluster of quiescent fibroblasts. The goal of this project is to investigate the molecular activation of quiescent cardiac fibroblast to myofibroblast based on the differential expression profile observed in human normal fibroblast, HF fibroblast, and myofibroblast. We will use ischemic heart model in mice to analyze expression of candidate proteins that correlated with the activation of myofibroblast after myocardial injury. The RNA inhibitor and overexpression strategies will be utilized to explore the mechanism of candidate genes in myofibroblast activation of primary cultured cardiac fibroblast.

The stipend for this SPUR project is funded by an American Heart Association grant awarded to Dr. Stavros Drakos, MD, PhD. The stipend for this project is $4,000 instead of $5,000 due to grant funding limitations.

Student Role

The student will assist in the procedures that generate ischemic injury in mouse model. The role of student includes collecting and processing the tissue specimens from animal models in the experiments. The responsibility will include tissue sectioning and immunofluorescence staining of control and ischemic myocardial tissue samples. The project activities will involve isolation and culture of primary fibroblast and gene specific modification. Since the project involves surgical procedure in mice, the student will assist with basic husbandry of the mice.

Student Learning Outcomes & Benefits

A great advantage of working in Dr. Drakos' lab specifically is that student will gain exceptional experiences in different areas of biomedical research, both basic and translational science. Student will have plenty of hands-on experiences in conducting laboratory research and learn a great deal about pathological fibrosis involvement in mechanisms of human disease. Since the student is required to present progress report in weekly lab meeting, he/she will have the learning experience in critical evaluation of the data, communication skills, and problem solving. The learning experience from this project will help and encourage the undergraduate student to prepare for research related career and professional development to become a scientist or physician devoted to evidence based medicine.

Stavros Drakos
Professor

Internal Medicine
School of Medicine

Technical skills:

The student will be assigned to work closely with a graduate student who is responsible for the research project and will initially assist the graduate student and technical specialist to perform the assays and experiments. The detail written protocols that are available in the laboratory will help guide the student to perform the assays required for experiments. The student is encouraged to operate on his/her own once he/she is proficient in performing the experiments. I will counsel the student occasionally to ensure that student understands the hypothesis being tested by experiments and the concluded outcome is correctly evaluated.

Communication skills:

Undergraduate trainee is required to participate in a weekly laboratory meeting to learn and discuss the research in progress in my laboratory. The trainee will assist and present the findings of project. I expect the student to develop the skill for data analysis, presentation preparation, and oral communication. The student will be required to present research publication related to the project that he/she involved in a monthly journal club meeting. My expectation is that student will learn the skills for literature search, scientific critique and reasoning, and communication in preparation for the required final presentation at the Undergraduate Research Summer Symposium.

The undergraduate trainee will be given increasing personal responsibilities and independence in performing the experiments, analysis, and presentation once the student has demonstrated under supervision that he/she proficiently developed those capabilities.