Beta Cell Formation in vivo Through Cellular Networking, Integration and Processing (CNIP) in Wild Type Adult Mice

Abstract

Insulin replacement therapy is essential in type 1 diabetic individuals and is required in ~40-50% of type 2 diabetics during their lifetime. Prior attempts at beta cell regeneration have relied upon pancreatic injury to induce beta cell proliferation, dedifferentiation and activation of the embryonic pathway, or stem cell replacement. We report an alternative method to transform adult non-stem (somatic) cells into pancreatic beta cells. The Cellular Networking, Integration and Processing (CNIP) approach targets cellular mechanisms involved in pancreatic function in the organ’s adult state and utilizes a synergistic mechanism that integrates three important levels of cellular regulation to induce beta cell formation: (i) glucose metabolism, (ii) membrane receptor function, and (iii) gene transcription. The aim of the present study was to induce pancreatic beta cell formation in vivo in adult animals without stem cells and without dedifferentiating cells to recapitulate the embryonic pathway as previously published (1-3). Our results employing CNIP demonstrate that: (i) insulin secreting cells can be generated in adult pancreatic tissue in vivo and circumvent the problem of generating endocrine (glucagon and somatostatin) cells that exert deleterious effects on glucose homeostasis, and (ii) long-term normalization of glucose tolerance and insulin secretion can be achieved in a wild type diabetic mouse model. The CNIP cocktail has the potential to be used as a preventative or therapeutic treatment or cure for both type 1 and type 2 diabetes.

A novel experimental model for human mixed acinar–ductal pancreatic cancer

“…We have previously shown that 48 h after the injection of Lentivirus coding for GFP is detected only in pancreatic tissues ( 14 ). We demonstrated previously using quantitative morphometric analysis of pancreatic transduction by the lentivirus vector (based on GFP expression) that 60% of the tissues expressed GFP ( 14 , 15 ) and this was confirmed in the present study with the expression of green fluorescent protein in the control group. Lentivirus vesicular stomatitis virus-glycoprotein envelope transduced both exocrine and endocrine tissues ( 14 , 15 ).…”

“…We demonstrated previously using quantitative morphometric analysis of pancreatic transduction by the lentivirus vector (based on GFP expression) that 60% of the tissues expressed GFP ( 14 , 15 ) and this was confirmed in the present study with the expression of green fluorescent protein in the control group. Lentivirus vesicular stomatitis virus-glycoprotein envelope transduced both exocrine and endocrine tissues ( 14 , 15 ). Importantly, expression was detected in the pancreas even after 4 weeks ( 14 , 15 ).…”

“…Lentivirus vesicular stomatitis virus-glycoprotein envelope transduced both exocrine and endocrine tissues ( 14 , 15 ). Importantly, expression was detected in the pancreas even after 4 weeks ( 14 , 15 ). The lentivirus vector expressed GFP was not found in any other tissues in the body, including heart, lung, liver, small intestine, brain, leg muscle and kidney by histology ( 14 , 15 ), and this was confirmed in the present study.…”

“…Importantly, expression was detected in the pancreas even after 4 weeks ( 14 , 15 ). The lentivirus vector expressed GFP was not found in any other tissues in the body, including heart, lung, liver, small intestine, brain, leg muscle and kidney by histology ( 14 , 15 ), and this was confirmed in the present study. Consistent with previous studies for our lab ( 14 , 15 ), no evidence of inflammation was observed in the present study with our method of lentivirus injection.…”

“…In contrast, we used wild-type adult mice, thus obviating the influence of the transgenic model, Cre protein toxicity and CRISPR/Cas9 off-target mutations to develop a model of pancreatic cancer in adult mice. To avoid these problems, we developed a novel approach, which utilizes viral injection into the adult mice pancreas ( 14 , 15 ) to recreate the pathophysiology of pancreatic cancer that develops in adulthood and thus avoids the development of compensatory mechanisms that occur when a gene is deleted during embryonic development.…”