PROJECT SUMMARY
Diabetes is characterized by the loss of functional pancreatic ß-cell mass. With the prevalence of diabetes
increasing across the globe, it is crucial that we develop a complete understanding of the gene regulatory
mechanisms governing the maintenance of insulin-producing ß-cell identity and function. In our previous work,
we demonstrated that the Isl1 transcription factor and its histone-modifying co-regulators Rnf20 and Rnf40 are
crucial for ß-cell gene expression. Recently, we observed that in mice, ß-cell-specific loss of Rnf20 (Rnf20¿ß-cell)
prompts a disruption of glucose homeostasis, loss of ß-cell identity gene expression, and an upregulation of cell
cycle inhibitor mRNA, p19Arf. In aging human and mouse ß-cells, cell cycle inhibitors encoded by Cdkn2a,
p19Arf and p16Ink4a, are upregulated and impart a reduction in functional capacity known as senescence. While
suppression of the age-associated upregulation in Cdkn2a expression is known to reverse ß-cell senescence,
the transcriptional mechanisms that repress Cdkn2a in healthy ß-cells are understudied. In this proposal, I will
investigate the regulatory relationship between Rnf20 and Cdkn2a to assess whether Rnf20 mediates ß-cell
senescence. My primary hypothesis is that Rnf20 protects against senescence-associated loss of ß-cell
functional identity through direct regulation of the Cdkn2a locus. I will address this hypothesis with two aims. In
Aim 1, I will evaluate how the ß-cell specific loss of Rnf20 affects the ß-cell cycle and the appearance of the
senescence-associated secretory phenotype. I will also perform systemic senolytic treatment to determine if
targeted elimination of senescent cells rescues the glucose intolerance and reduced plasma insulin levels that
occur in Rnf20¿ß-cell mice. In Aim 2, I will determine whether Rnf20 regulates the transcription of Cdkn2a. I will
perform Chromatin Immunoprecipitation (ChIP) to assess if Rnf20 directly occupies the Cdkn2a promoter and
ATAC-seq to see if loss of Rnf20 increases Cdkn2a chromatin accessibility. Our results will increase
understanding of the transcriptional mechanisms regulating both the maintenance of ß-cell identity and the
process of ß-cell senescence. Additionally, our outcomes may be used to inform future diabetes therapies
through either the incorporation of Rnf20 into either ß-cell differentiation protocols or as a biomarker for ß-cell
health. Furthermore, the proposed research plan provides a balanced training platform wherein I will expand
both my critical thinking and technical skillset through my investigations of novel regulatory mechanisms of ß-
cell homeostasis. As a trainee at the University of Alabama at Birmingham (UAB) and in UAB’s Diabetes
Research Center, I have access to a strong community of supportive faculty members to assist in the successful
completion of the proposed study. Taken together, this research and training plan is a fundamental catalyst of
my trajectory toward becoming an independent research scientist.
