title: Failure to achieve normal metabolic response in non-obese diabetic mice and streptozotocin-induced diabetic mice after transplantation of primary murine hepatocytes electroporated with the human proinsulin gene (p3MTChins).
abstract: BACKGROUND
  A recent study by Chen et al described a therapy for diabetes that involved electroporation of primary hepatocytes with human proinsulin cDNA, p3MTChins. Intrahepatic transplantation of treated hepatocytes into streptozotocin (STZ) murine and porcine models led to euglycemia, weight maintenance, and normal insulin production. We tested the repeatability of their basic experiments and transplantation technique and expanded the study to include an autoimmune model.
  
  
  METHODS
  Hepatocytes were isolated from B6 mice, electroporated with p3MTChins, and glucose-challenged or were injected into hepatic or spleen parenchyma of STZ-diabetic B6 and non-obese diabetic mice. Outcomes included survival, serum glucose levels, insulin, and c-peptide release. Untransfected primary hepatocytes and mice transplanted with these cells served as controls.
  
  
  RESULTS
  p3MTChins-hepatocytes secreted insulin during glucose challenge, but glucose levels did not change with increasing glucose concentrations. Direct hepatic injection led to high mortality rates. Mice that underwent intrasplenic injection survived for >50 days (control = 4 days) and had a mild but stable improvement in hyperglycemia. C-peptide in both mouse models was detectable but eventually declined to baseline in the non-obese diabetic mice.
  
  
  CONCLUSIONS
  Hepatocytes can be transfected with p3MTChins to produce human insulin but may lack the proper glucose-sensing or complex storage and secretion capabilities that allow for a finely tuned dynamic insulin response. Treatment is subtherapeutic, and p3MTChins-hepatocyte function may not endure in an autoimmune model. Without successful preliminary findings, cell therapy involving electroporation of p3MTChins does not appear to be practical as a therapy for diabetes and may not be a strategy to pursue at this time.
authors.affiliation: Department of Surgery, University of California-San Francisco, San Francisco, California, USA. Electronic address: ryalee@alamedahealthsystem.org.
authors.forename: R H
authors.initials: RH
authors.lastname: Lee
authors.affiliation: Department of Surgery, University of California-San Francisco, San Francisco, California, USA.
authors.forename: G
authors.initials: G
authors.lastname: Roll
authors.affiliation: Department of Surgery, University of California-San Francisco, San Francisco, California, USA.
authors.forename: V
authors.initials: V
authors.lastname: Nguyen
authors.affiliation: Department of Surgery, University of California-San Francisco, San Francisco, California, USA.
authors.forename: H
authors.initials: H
authors.lastname: Willenbring
authors.affiliation: Department of Surgery, University of California-San Francisco, San Francisco, California, USA.
authors.forename: Q
authors.initials: Q
authors.lastname: Tang
authors.affiliation: Department of Surgery, University of California-San Francisco, San Francisco, California, USA.
authors.forename: S-M
authors.initials: SM
authors.lastname: Kang
authors.affiliation: Department of Surgery, University of California-San Francisco, San Francisco, California, USA.
authors.forename: P G
authors.initials: PG
authors.lastname: Stock
mesh_terms: D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D002096:C-Peptide / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D003921:Diabetes Mellitus, Experimental / Q000209:etiology / Q000378:metabolism* / Q000628:therapy*; D018274:Electroporation; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D022781:Hepatocytes / Q000637:transplantation*; D006801:Humans; D007328:Insulin / Q000378:metabolism; D008099:Liver; D008297:Male; D051379:Mice; D016688:Mice, Inbred NOD; D011384:Proinsulin / Q000235:genetics*; D013154:Spleen; D013311:Streptozocin; D013552:Swine
chemical_list: D001786:Blood Glucose; D002096:C-Peptide; D018076:DNA, Complementary; D007328:Insulin; D013311:Streptozocin; D011384:Proinsulin