Research Projects

India is the diabetes capital of the world. T1DM is an autoimmune disease wherein the immune system erroneously targets and destroys the insulin-secreting beta (β) cells of the pancreas, preventing adequate insulin production, resulting in high glucose levels, ketoacidosis, and potentially fatal complications. T1DM patients require daily insulin injections to maintain their blood sugar and experience pain and distress due to the lifelong administration of these injections. The field needs alternative approaches and innovative β-cell replacement therapy. Encouraging preclinical animal studies indicate that induced pluripotent stem cells (iPSCs) can reverse diabetes, but lacked β-cell functionality and is termed “β-cell-like” cells. As a novel stem cell source Intestinal Stem Cell (ISC) can be a calculative bargain. The intestine and pancreas share a close ontogenetic link that makes their interconversion potential highly anticipated. A few proof-of-principle experiments suggest that gut enterochromaffin cells can be converted to β-cells. To address this research gap, we will use a unique strategy to alter the plasticity of human ISCs to insulin-producing pancreatic endocrine lineage, providing an autologous alternative supply of insulin-producing β-cells. Based on our preliminary data, we are proposing that intestinal stem cells of mice/humans can be differentiated into glucose-responsive, insulin-producing β-cells by altering the epigenetic marks (by deleting DNMT1) and modulating Wnt/β-catenin signaling in a stage-specific manner. We hypothesize that DNMT1 inhibition and stepwise Wnt modulation will create a permissive chromatin environment to facilitate reprogramming of ISC to pancreatic progenitor, and then to pancreatic endocrine lineage and finally to β-cells. As a model system we will use the “Enteroid”/organoid model-derived from primary mice/human intestinal crypts, and we propose to delete DNMT1 using CRISPR/Cas genome editing technology or by using specific inhibitors (Aza-5, GSK-3484862) and stage-specific Wnt/β-catenin modulation. We will use RNA-Seq, confocal immunofluorescence, methylome studies, and qPCR to confirm the β-cell lineage reprogramming. We will also use the STZ-treated diabetic mice model for in vivo enteroid transplant to rescue diabetes. NOD/Scid mice will be used for human enteroid differentiation and β-cell transplantation. As an outcome our findings will raise the possibility of using DNMT1 inhibition and Wnt-modulated of ISCs as a source of insulin-producing cells to treat TIDM. Upon success, we will be the first to identify the ISC and their potential in β-cell reprogramming and contribute to developing a viable diabetes therapy. Our application is distinctly innovative, as we propose to use mice ISCs/patient-derived autologous ISCs and cutting-edge organoid models. Reprogramming autologous human ISCs to create native glucose-responsive insulin-secreting β-cells is a highly novel and translational approach.