AGE REVERSAL BY TRANSCRIPTOMIC REPROGRAMMING

Abstract

Aging is a progressive multifaceted functional decline of a biological system. Chronic age-related conditions such as cancer, cardiovascular, and neurodegenerative diseases are leading causes of death worldwide. As the elderly population is experiencing an exponential growth, aging is becoming a pressing problem for our society. To address this global challenge, there is a need for novel, safe, and effective rejuvenation therapies aimed at reversing age-related phenotypes and improving human health. In my dissertation work, I sought to advance the discovery of age reversal therapies by developing novel cell engineering tools and applying them to the aging problem. With gene expression being a key determinant of cell identity and function, I have focused my efforts on reprogramming the cell transcriptome to a youthful state. To this end, we first developed an integrated pipeline that enables fast and efficient mammalian cell engineering using algorithms for target identification, modular tools for library cloning, and next-generation sequencing-coupled readouts. Next, we built a functionally interpretable RNA clock as an integrative aging assay that can accurately predict the age of human fibroblasts, as well as the effect of several pro- and anti- aging interventions. Lastly, we used our cell engineering pipeline along with our aging clock to perform the first age reversal screen in primary human cells. Our work uncovered several targets for cellular rejuvenation that induced younger transcriptomes in middle-aged and old human fibroblasts. The most promising of these targets was serine and arginine rich splicing factor 1 (SRSF1), whose overexpression had a robust rejuvenating effect on both the transcriptome and function of aged human fibroblasts. Throughout these studies I propose a new paradigm for the discovery of age reversal interventions by using transcriptomic reprogramming screens.