Understanding cellular and transcriptional regulatory changes in human aging.

摘要

ABSTRACT In this project, we propose to add Ms. Marliette Rodriguez Matos as a central member in our project to study the aging of CD4+ T lymphocytes. The parent R01 represents an ambitious project to study a primary, purified human cell type that has been associated with age-related diseases and has been shown to manifest the systematic changes in DNA methylation that occur with age in multiple tissue types and species. Our goal is to understand how much of the spectrum of changes of molecular genomic properties with age are due to cell subtype changes, how many are cell-autonomous, how these reflect cell signalling effects on transcription factor biology, and how inter-individual DNA sequence variation interacts with these molecular and cellular phenotypes. Towards these goals we are performing genotyping, T cell receptor amplicon sequencing, and chromatin accessibility studies using ATAC-seq on the samples. Our original plan was to add bulk RNA-seq and DNA methylation studies, but we are now exploring instead a single cell RNA-seq approach with pseudobulking, and are concerned that the DNA methylation studies will be uninformative given more recent data that indicates these changes represent a decrease in multipotent stem cell proportions in tissues with age, which may explain why the DNA methylation clock has not been demonstrated to work in purified cells. We have purified our cells with the CD4+ surface marker, which depletes all precursor cells. Our ongoing work led by Dr. Katherine Crocker is designed to shed light on this issue so that we make our experimental investments wisely. Ms. Rodriguez Matos will take the lead in performing the chosen set of molecular genomic assays on the cohort of 400 samples. She comes to our group with strong wet bench experience, and has proven herself to be exceptional technically with genomic assays. Given the extensive project delays enforced by the COVID-19 pandemic, adding Ms. Rodriguez Matos is based on the practical consideration that this will allow us to deliver the genomic data needed for the final, analytical stage of the project. We will develop Ms. Rodriguez Matos’ participation in this project as part of a structured training experience. We describe in this proposal how her project activities will involve activities that are designed to foster transferrable skills. These skills will not only be related to the field of aging but also project management, communication, leadership, and supervisory skills, and training in ethics and integrity. Her analytical skills will be fostered by working with the group of co-PI Dr. Tuuli Lappalainen at the New York Genome Center, rounding out her skills. Ms. Rodriguez Matos’ has defined her career goal explicitly as a future independent investigator. Our goal during the R01 project is to give her the skills and publications that will give the foundation needed for that outcome. We would welcome the opportunity to train a Hispanic-American woman as a future leader in human genomics and in the field of aging research in particular.