title: Hydrodynamics-based transfection of the combination of betacellulin and neurogenic differentiation 1 DNA ameliorates hyperglycemia in mice with streptozotocin-induced diabetes.
abstract: BACKGROUND
  The biohazards caused by the viral delivery of pancreatic transcription factors, including neurogenic differentiation 1 (Neurod1) and Betacellulin (Btc), to the murine liver limit application of this procedure in reversing diabetes. We aimed to evaluate the feasibility of hydrodynamics-based transfection (HBT) with Neurod1 and Btc in improving hyperglycemia.
  
  
  METHODS
  Murine hepatocellular carcinoma (Hepa1-6) cells were transfected with the combination of Neurod1-expressing plasmid, pcDNA3.1/V5-His A (pcDNA)-Neurod1, and Btc-expressing plasmid, pcDNA3.1/V5-His A (pcDNA)-Btc. Hepatic delivery of a combination of pcDNA-Neurod1 and pcDNA-Btc (experimental group) or pcDNA (control group) to mice with streptozocin-induced diabetes was achieved by HBT. The sequential serum glucose and alanine aminotransferase (ALT) levels were assessed.
  
  
  RESULTS
  On day 3 after transfection, the transfection efficiencies of pcDNA-Btc and pcDNA-Neurod1 in the Hepa1-6 cells were 20% and 8%, respectively; respective values in the mouse livers were 30% and 10%. At 1 week after HBT, aside from hepatic expression of insulin, the experimental mice had a significantly lower sugar level (8-14 days after HBT, P values ranged from 0.034 to <0.001) than the control mice; the difference remained for 1 week but diminished afterward. The ALT levels and the body weight change were not different between the two groups. No mortality was noted in both groups.
  
  
  CONCLUSIONS
  The hypoglycemic effect of Neurod1 and Btc delivered by HBT was transient and associated with negligible complications. In studies on the short-term hypoglycemic effects of Neurod1 and Btc in vivo, HBT is a potential alternative to viral delivery of Neurod1 and Btc to the murine liver.
authors.affiliation: Liver Research Center and Department of Hepatogastroenterology, Chang Gung Memorial Hospital, College of Medicine, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Chang-Mu
authors.initials: CM
authors.lastname: Sung
authors.forename: Chau-Ting
authors.initials: CT
authors.lastname: Yeh
authors.forename: Shuen-Shian
authors.initials: SS
authors.lastname: Shiau
authors.forename: Chun-Kai
authors.initials: CK
authors.lastname: Liang
authors.forename: Ming-Ling
authors.initials: ML
authors.lastname: Chang
mesh_terms: D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000008:administration & dosage / Q000235:genetics / Q000627:therapeutic use*; D066259:Betacellulin; D002460:Cell Line; D003922:Diabetes Mellitus, Type 1 / Q000139:chemically induced / Q000378:metabolism / Q000628:therapy*; D005240:Feasibility Studies; D005260:Female; D015316:Genetic Therapy / Q000009:adverse effects / Q000379:methods; D006439:Hemodynamics; D022781:Hepatocytes / Q000378:metabolism / Q000473:pathology; D057446:Hydrodynamics; D006943:Hyperglycemia / Q000517:prevention & control*; D007275:Injections, Intravenous; D007328:Insulin / Q000096:biosynthesis / Q000097:blood; D036341:Intercellular Signaling Peptides and Proteins / Q000008:administration & dosage / Q000235:genetics / Q000627:therapeutic use*; D008297:Male; D051379:Mice; D010957:Plasmids / Q000008:administration & dosage* / Q000009:adverse effects / Q000627:therapeutic use; D013311:Streptozocin / Q000633:toxicity; D013997:Time Factors; D014162:Transfection / Q000379:methods*
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D066259:Betacellulin; D007328:Insulin; D036341:Intercellular Signaling Peptides and Proteins; D013311:Streptozocin; C587852:Btc protein, mouse; C501008:Neurod1 protein, mouse