SPUR 2021: Vascular ceramide biosynthesis disrupts endothelial cell barrier function and contributes to cognitive impairment in mice with and without ischemic stroke, Defining the contribution from endothelial cell autophagy and PKC activation to vascular cognitive impairment, Characterizing metabolism in endothelial cells obtained from healthy individuals and patients with cardiovascular disease

Background

Project 1. Vascular dysfunction contributes to cognitive decline. Our lab has shown that ceramide accrual in preclinical models of type 2 diabetes (T2D) leads to vascular dysfunction. It is unknown whether vascular ceramide biosynthesis leads to cognitive decline in preclinical models of T2D and / or exacerbates cognitive decline in preclinical models of T2D upon exposure to ischemic stroke. We will use novel, vascular specific, "gain of ceramide" and "loss of ceramide" murine models to explore this.

Project 2. Cardiovascular diseases affect 80% of persons > 65. Cognitive decline is associated with an aging endothelial cell phenotype, but the mechanisms are unknown. Repressed endothelial cell autophagy mimics an aging phenotype and we will determine whether this is sufficient to evoke cognitive decline in young mice. Importantly, we will assess the potential for a novel small molecule - bryostatin - to correct EC homeostasis to an extent that maintains cognitive function in older mice.

Project 3. We obtain endothelial cells (ECs) that line the arteries of healthy individuals and patients with cardiovascular disease. After verifying the purity of the ECs, we treat them with a variety of agonists and antagonists to evaluate their integrity. Next, we use unbiased discovery approaches to discern signaling pathways that might be upregulated or downregulated in the context of aging, hypertension, diabetes, heart failure.

Student Role

Students play an extremely important role in every experiment. Past student involvement has included:

  1. experimental set-up;
  2. assessing arterial function;
  3. quantifying intravascular signaling pathways;
  4. metabolic phenotyping;
  5. sectioning and staining brain tissue for infarction, neuroinflammation, neurodegeneration, ceramide accrual;
  6. animal husbandry;
  7. cognitive assessment of mice;
  8. data collection, reduction, analysis, interpretation, and presentation;
  9. flow cytometry;
  10. qRT PCR, western blotting; and
  11. evaluating subject and patient records.

Student Learning Outcomes & Benefits

Learning outcomes. Students will learn:

  1. the daily routines of an academic researcher, MS student, PhD student, Postdoctoral Scholar, and Junior faculty member;
  2. cardiovascular physiology in general, and vascular biology in particular;
  3. the scientific method;
  4. various laboratory tests / procedures;
  5. how to critically evaluate data that are being collected;
  6. organization and time management skills;
  7. how to make an effective data presentation in oral and / or poster format.

Benefits. If you choose to go on to one of the pre-health professions, and your interviewer asks "tell me about your research" ... you will be able provide a succinct answer that includes:

  1. the background/rationale/clinical relevance of your topic;
  2. your directional hypothesis(es);
  3. methods you used;
  4. main results you obtained;
  5. how your findings synthesize with what is known currently; and
  6. how you moved the field forward in a manner that was NOT incremental. Furthermore, it is hoped that you will be able to tell them that you presented your findings at a local, national, and / or international conference. We will help you to accomplish this.

Remote Contingency Plan

Remote undergraduate research internship contingency plans have been in effect since March 2020. Examples include : 1) vascular data reduction, entry, analysis, and statistical evaluation; 2) histology imaging, analysis. and evaluation; 3) subject and patient record interrogation and organization. All students attend lab meetings and seminars via zoom when their schedule permits, and one-on-one meetings with the PI when their schedule permits. Another potential scenario is restricted laboratory access vs. completely remote. We have implemented this successfully as well. Students come to lab on pre arranged schedules, to facilitate physical distancing and minimize laboratory density. Students have been trained and completed experiments in this mannerl.

J David Symons
Professor

Nutrition & Integrative Physiology
College of Health
Molecular Biology Program

Philosophy. I try to give the student sufficient knowledge, training, and resources to foster success. It is up to them to seize the opportunity to elevate their level of understanding concerning a certain topic e.g., the role of the vasculature in contributing to cognitive decline in the context of type 2 diabetes.

Mentoring activities. Multi PI lab meeting are held Monday AM. My lab meeting is Wed AM. Seminars in Metabolism is Thurs PM. Other seminars are attended based on topic e.g,. Cardiology, Nephrology, Molecular Medicine Chalk Talk. I perform experiments with the students and therefore am in the lab daily for one-on-one. Each student will present their particular data set at least twice during the summer. I will directly assist with poster and / or oral presentation. I will strongly encourage presentation of data at SPUR summer symposium, UU URS, UCUR, NCUR. Undergraduate students from my lab have routinely presented their work at International Conferences and have coauthored publications.