Poster V11 - Reprogramming adipocytes to produce Insulin as an alternative therapy in Diabetes.

Introduction: Cell reprogramming is a promising strategy for the treatment of various diseases involving the loss of cell function, such as diabetes mellitus (DM). DM is a chronic metabolic disease with an increasingly high incidence that is projected to double in the next 30 years (ONG et al., 2023). Therefore, there is a need to explore novel, large-scale, and reliable methods for insulin production to address the growing demand required to treat DM. Moreover, new therapies are necessary to overcome the limitations and deficiencies inherent in the existing standard-of-care treatment, such as the deficiency of beta cells and suboptimal control of blood glucose levels in patients with DM. Accordingly, there is great importance in methods that are capable of restoring beta cell functionality as a more efficient alternative for the treatment of DM, especially type 1 DM. Thus, the capacity to create cells that produce insulin by utilizing modern reprogramming techniques offers a reliable and innovative solution for insulin production and treatment of DM

Materials and

Methods: Here we explore the potential for mouse embryonic fibroblast cell line (3T3L1), a widely recognized model of adipocytes, to be reprogrammed to insulin producing cells (iPCs) using different cocktails of transcription factors associated with insulin production in beta cells, Pdx1, Ngn3, Mafa, NeuroD1, Pax4, FoxA2, Nkx6.1, TCF3, Trp63 and Sox9

Results, Conclusions, and Discussions: At 3 days post-transfection, cells were stimulated with trypsin to produce aggregates to effectively simulate the microenvironment of beta cells in the pancreatic islets. At 4 days post-transfection, our results showed that the cells formed clusters similar to islets within pancreas, as confirmed visually by microscopy. Moreover, we observed a significant three fold increase in insulin gene expression in cells treated with Pdx1, Ngn3, Mafa TCF3 and Pax4 compared to the sham transfected control (p=0.01). Taken together, our results indicate that our methods represent a new avenue for developing iPCs, or cells similar to beta cells, that are capable of producing insulin, highlighting the potential of this novel approach for the transdifferentiation of adipocytes to iPCs for the treatment of DM.

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