Reversal of diabetes following transplantation of an insulin-secreting human liver cell line: Melligen cells

Abstract

As an alternative to the transplantation of islets, a human liver cell line has been genetically engineered to reverse type 1 diabetes (TID). The initial liver cell line (Huh7ins) commenced secretion of insulin in response to a glucose concentration of 2.5 mmol/l. After transfection of the Huh7ins cells with human islet glucokinase, the resultant Melligen cells secreted insulin in response to glucose within the physiological range; commencing at 4.25 mmol/l. Melligen cells exhibited increased glucokinase enzymatic activity in response to physiological glucose concentrations, as compared with Huh7ins cells. When transplanted into diabetic immunoincompetent mice, Melligen cells restored normoglycemia. Quantitative real-time polymerase chain reaction (qRT-PCR) revealed that both cell lines expressed a range of β-cell transcription factors and pancreatic hormones. Exposure of Melligen and Huh7ins cells to proinflammatory cytokines (TNF-α, IL-1β, and IFN-γ) affected neither their viability nor their ability to secrete insulin to glucose. Gene expression (microarray and qRT-PCR) analyses indicated the survival of Melligen cells in the presence of known β-cell cytotoxins was associated with the expression of NF-κB and antiapoptotic genes (such as BIRC3). This study describes the successful generation of an artificial β-cell line, which, if encapsulated to avoid allograft rejection, may offer a clinically applicable cure for T1D.

Genomic Characterization of Metformin Hepatic Response

“…Interestingly, reducing the hepatic eIF2α signaling pathway in mice was shown to lead to reduced hepatic glucose production through reduced hepatic gluconeogenic gene expression [39]. BIRC3 codes for the protein baculoviral IAP repeat containing 3, and its expression was associated with the survival of insulin-secreting human liver cell line, which restored normoglycemia when transplanted into diabetic immunoincompetent mice [40]. Furthermore, PCK1 encodes the phosphoenolpyruvate carboxykinase 1 (PEPCK) enzyme, which catalyzes the conversion of oxaloacetate to phosphoenolpyruvate and carbon dioxide and is considered a key pathway for hepatic gluconeogenesis [41].…”

Antigen-Encoding Bone Marrow Terminates Islet-Directed Memory CD8+ T-Cell Responses to Alleviate Islet Transplant Rejection

“…With significant advances made recently in generation of stem cell-derived b-cells (42,43) and genetically engineered insulin-secreting cells (44,45), it is conceivable that generation of personalized insulin-secreting cells will be feasible in the future. However, persistent anti-islet memory T-cell responses represent a threat to clinical application of such approaches.…”

“…from non-b-cells such as hepatocytes (44,45). However, prevention of T1D progression in at-risk or recent-onset human subjects may require a more complex approach.…”

Bioengineering Priorities on a Path to Ending Organ Shortage

“…For example, pancreatic islet transplants delivered into the liver can function, but do not replicate the microenvironmental pancreas map. Human liver cells can also be engineered to reverse type 1 diabetes [14]. It remains to be determined if these approaches will provide long-term correction, or if these organs may become impaired over time, with deleterious effects to the patient.…”