Partial pancreatic transdifferentiation of primary human hepatocytes in the livers of a humanised mouse model

Abstract

These data show that human hepatocytes can be induced to undergo partial pancreatic transdifferentiation in vivo, indicating that the technology holds promise for the treatment of type 1 diabetes.

Use of a Hybrid Adeno-Associated Viral Vector Transposon System to Deliver the Insulin Gene to Diabetic NOD Mice

“…Additionally, there will be no requirement for the ex vivo manipulation of a patient’s liver, and subsequent adoptive transfer of artificial β cells. Hepatocytes have been shown to be suitable candidates for the generation of artificial β cells [ 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 ]. In most cases, hyperglycaemia has been ameliorated by the expression of various β-cell transcription factors in liver cells, delivered most commonly by adenoviral vectors [ 13 , 16 , 17 , 19 ].…”

“…Our laboratory has established that the dual expression of insulin and specific β-cell transcription factors in liver cell lines and primary hepatocytes has a synergistic effect causing pancreatic transdifferentiation, storage of insulin in granules, regulated insulin secretion to glucose and other β-cell secretogogues, and, most importantly, the ability to permanently reverse diabetes [ 9 , 10 , 20 , 25 ]. In order to obtain consistently high transduction rates and transgene isolation to the liver in diabetic animals, we developed a novel microsurgical procedure of intervallic infusion in full flow occlusion (FFO) to deliver a lentiviral vector.…”

“…These results are likely attributable to the FFO technique, eliminating significant quantities of blood that would have rapidly inactivated the vector by a complement-mediated mechanism. In a world first, we have also observed the expression of human β-cell transcription factors in primary human hepatocytes engrafted into the liver of the humanised FRG mouse [ 25 ], in which chimeric human/mouse livers can be generated [ 26 , 27 ].…”

“…In previous successful studies, the lentiviral vector system stably incorporated the INS-FUR gene into liver cells [ 9 , 10 , 11 , 25 ]. In an attempt to investigate whether it was the episomal expression of INS - FUR using the AAV8 system that resulted in the absence of pancreatic transdifferentiation and persistence of abnormal glucose tolerance, we also employed the AAV8/ piggyBac system, which can mediate the transposition of transgenes into the host genome [ 33 , 34 , 35 ].…”

“…In an attempt to investigate whether it was the episomal expression of INS - FUR using the AAV8 system that resulted in the absence of pancreatic transdifferentiation and persistence of abnormal glucose tolerance, we also employed the AAV8/ piggyBac system, which can mediate the transposition of transgenes into the host genome [ 33 , 34 , 35 ]. As the piggyBac system has shown sustained gene expression in adult mice [ 35 ], we hypothesised that the AAV8- INS-FUR - piggyBac system would result in somatic integration and long-term gene expression, therby reversing their diabetes in a similar manner to the lentiviral system [ 9 , 10 , 25 ]. Expression of INS-FUR resulted in euglycaemia and normal IPGTTs in the mice that received AAV8/ piggyBac - INS-FUR and these were subsequently subjected to FFO surgery.…”