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: Chang-Mu Sung, Chau-Ting Yeh, Shuen-Shian Shiau, Chun-Kai Liang, Ming-Ling Chang
• Journal: Diabetes technology & therapeutics (Diabetes Technol Ther) Vol. 13 Issue 5 Pg. 519-25 (May 2011) ISSN: 1557-8593 [Electronic] United States
• PMID: 21406008 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Basic Helix-Loop-Helix Transcription Factors
• Betacellulin
• Btc protein, mouse
• Insulin
• Intercellular Signaling Peptides and Proteins
• Neurod1 protein, mouse
• Streptozocin

Topics

• Animals
• Basic Helix-Loop-Helix Transcription Factors (administration & dosage, genetics, therapeutic use)
• Betacellulin
• Cell Line
• Diabetes Mellitus, Type 1 (chemically induced, metabolism, therapy)
• Feasibility Studies
• Female
• Genetic Therapy (adverse effects, methods)
• Hemodynamics
• Hepatocytes (metabolism, pathology)
• Hydrodynamics
• Hyperglycemia (prevention & control)
• Injections, Intravenous
• Insulin (biosynthesis, blood)
• Intercellular Signaling Peptides and Proteins (administration & dosage, genetics, therapeutic use)
• Male
• Mice
• Plasmids (administration & dosage, adverse effects, therapeutic use)
• Streptozocin (toxicity)
• Time Factors
• Transfection (methods)