Assessing the Comparative Effectiveness of Upregulation of Beta Cell Identity Genes and Downregulation of Senescence-Associated Markers for Senescence Reversal: A Research Protocol

Abstract

Introduction: Type 1 diabetes (T1D) is a chronic condition characterized by the immune-mediated destruction of pancreatic beta cells, resulting in insulin deficiency and elevated blood glucose levels. Despite substantial advancements in understanding the pathogenesis, epidemiology and management of T1D, a treatment for the disease is yet to be discovered. Diabetic environment appears to trigger cellular senescence in a subset of beta cells resulting in proliferation arrest and drastic phenotypic and genotypic changes in these cells. It has been suggested that senescent beta cells may exacerbate T1D progression by establishing a senescence-associated secretory profile (SASP), contributing to chronic inflammation and tissue dysfunction. Despite the potential therapeutic significance of overturning senescence, the optimal approach for such intervention is largely overlooked. This proposal, therefore, aims to unveil the relative effectiveness of two predominant strategies in senescence reversal.

Methods: Senescence is induced through UV irradiation and doxorubicin treatment in beta cells, extracted from pancreases of male and female NOD mouse models. Lipid Nanoparticle (LNP) delivery is subsequently used to overexpress two beta-cell identity genes and underexpress two senescence markers in different conditions. Changes in the expression of the predominant SASP factors, IL-6, IL-8, and TNF-α are measured and compared through Nanostring technology and one-way ANOVA, respectively. This could quantify the absolute and relative effectiveness of the aforementioned strategies in senescence reversal.

Discussion: Both reversal mechanisms are anticipated to successfully overturn senescence, which can be indicated by a significant decrease in the mean levels of SASP factors post-treatment. Nevertheless, downregulation of senescence markers may be the more effective of the two, yielding more substantial results with regards to senescence reversal. It directly addresses the issue of senescence without overwhelming cellular machinery. In addition, fewer compensatory mechanisms seem to be associated with senescence-associated genes, hence the treatments are expected to be longer lasting.

Conclusion: Results of this study can contribute to the development of therapeutic regimens for diabetes prevention and reversal. Moreover, these findings have broad applicability across various contexts where senescence reversal is of value.