Reprogramming of human pancreatic exocrine cells to β-like cells

Abstract

Rodent acinar cells exhibit a remarkable plasticity as they can transdifferentiate to duct-, hepatocyte- and islet β-like cells. We evaluated whether exocrine cells from adult human pancreas can similarly respond to proendocrine stimuli. Exocrine cells from adult human pancreas were transduced directly with lentiviruses expressing activated MAPK (mitogen-activated protein kinase) and STAT3 (signal transducer and activator of transcription 3) and cultured as monolayers or as 3D structures. Expression of STAT3 and MAPK in human exocrine cells activated expression of the proendocrine factor neurogenin 3 in 50% to 80% of transduced exocrine cells. However, the number of insulin-positive cells increased only in the exocrine cells grown initially in suspension before 3D culture. Lineage tracing identified human acinar cells as the source of Ngn3- and insulin-expressing cells. Long-term engraftment into immunocompromised mice increased the efficiency of reprogramming to insulin-positive cells. Our data demonstrate that exocrine cells from human pancreas can be reprogrammed to transplantable insulin-producing cells that acquire functionality. Given the large number of exocrine cells in a donor pancreas, this approach presents a novel strategy to expand cell therapy in type 1 diabetes.

Pancreas regeneration

“…Changes in α-cell to β-cell ratios and the appearance of cells positive for both glucagon and insulin have provided some evidence for such conversions; however, in the absence of lineage tracing, direct evidence is still lacking. Other cell types such as pancreatic acinar cells, ductal cells, gall bladder cells, and intestinal cells have also been used to generate insulin-expressing cells, but these cells did not form long-term stable grafts after transplantation, suggesting incomplete cell fate conversion or an unstable epigenetic state 121–124 . At this point, the main challenge in translating approach into the clinic is to define reliable methods for efficient production of human β-like cells that can develop stable and functional transplants.…”

β-cell replacement sources for type 1 diabetes: a focus on pancreatic ductal cells

“…Recent developments in their work showed the potential to induce NGN3 expression in human exocrine cells by lentiviral-induced overexpression of mitogenactivated protein kinase, and signal transducer and activator of transcription 3 [Lemper et al 2015]. The NGN3-expressing cells entered an endocrine program and specified into functional β cells 42 days after transplantation into normoglycemic immunocompromised mice.…”

<i>In vivo</i> direct reprogramming of liver cells to insulin producing cells by virus-free overexpression of defined factors

“…The reprogrammed cells can significantly improve the STZ diabetic mice symptoms of hyperglycemia, and insulin secretion can reach 23% of the normal insulin secretion level. Currently, adenoviral vectors have been widely used in the reprogramming system for exogenous gene transfer because of its high transfection efficiency [22][23][24][25][26][27], but still face the risks of integration and immune rejection. Studies have shown that up to 2×10…”

“…The efficiency of luciferase plasmid by hydrodynamic delivery was assessed by in vivo bioluminescence imaging of luciferase activity using an IVIS Spectrum system (Perkin-Elmer, Waltham, MA) at 3,6,12,24,48, and 72 hours post-injection. Mice were anesthetized with isoflurane, followed by intraperitoneally injection of 150 mg/kg D-luciferin (Goldbio, USA).…”