Reprogramming of Human Pancreatic Organoid Cells into Insulin-Producing β-Like Cells by Small Molecules and in Vitro Transcribed Modified mRNA Encoding Neurogenin 3 Transcription Factor

摘要

Reprogramming of non-endocrine pancreatic cells into insulin-producing cells represents apromising therapeutic approach for the restorationof endogenous insulin production in diabetic patients. In this paper, we report that human organoidcells derived from the pancreatic tissue can be reprogrammed into the insulin-producing cells (IPCs) bythe combination of in vitro transcribed modifiedmRNA encoding transcription factor neurogenin 3and small molecules modulating the epigenetic stateand signalling pathways. Upon the reprogramming,IPCs formed 4.6 ± 1.2 % of the total cells and expressed typical markers (insulin, glucokinase, ABCC8,KCNJ11, SLC2A2, SLC30A8) and transcription factors (PDX1, NEUROD1, MAFA, NKX2.2, NKX6.1,PAX4, PAX6) needed for the proper function of pancreatic β-cells. Additionally, we have revealed a positive effect of ALK5 inhibitor RepSox on the overallreprogramming efficiency. However, the reprogrammed IPCs possessed only a partial insulin-secretory capacity, as they were not able to respond tothe changes in the extracellular glucose concentration by increasing insulin secretion. Based on theachieved results we conclude that due to the incomplete reprogramming, the IPCs have immature character and only partial properties of native humanβ-cells.