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https://doi.org/10.1371/journal.pone.0001734
Title: | Nonvirally modified autologous primary hepatocytes correct diabetes and prevent target organ injury in a large preclinical model | Authors: | Chen N.K.F. Wong J.S. Kee I.H.C. Lai S.H. Thng C.H. Ng W.H. Ng R.T.H. Tan S.Y. Lee S.Y. Tan M.E.H. Sivalingam J. Chow P.K.H. Kon O.L. |
Keywords: | C peptide complementary DNA glucose insulin messenger RNA transcriptome C peptide insulin messenger RNA streptozocin animal cell animal experiment animal model animal tissue aorta article cell isolation cell transplantation controlled study diabetes mellitus dyslipidemia electroporation ex vivo study expression vector eye female gene transfer glucose blood level glucose intolerance hyperglycemia kidney liver liver cell liver function male metabolic disorder nonhuman organ injury plasmid streptozocin diabetes swine animal DNA microarray experimental diabetes mellitus gene expression profiling gene therapy genetics glucose tolerance test human injury metabolism pancreas islet beta cell pathology radioimmunoassay retina reverse transcription polymerase chain reaction secretion transplantation Suidae Animals Aorta Blood Glucose C-Peptide Diabetes Mellitus, Experimental Electroporation Gene Expression Profiling Gene Therapy Gene Transfer Techniques Glucose Tolerance Test Hepatocytes Humans Insulin Insulin-Secreting Cells Kidney Liver Oligonucleotide Array Sequence Analysis Radioimmunoassay Retina Reverse Transcriptase Polymerase Chain Reaction RNA, Messenger Streptozocin Swine |
Issue Date: | 2008 | Citation: | Chen N.K.F., Wong J.S., Kee I.H.C., Lai S.H., Thng C.H., Ng W.H., Ng R.T.H., Tan S.Y., Lee S.Y., Tan M.E.H., Sivalingam J., Chow P.K.H., Kon O.L. (2008). Nonvirally modified autologous primary hepatocytes correct diabetes and prevent target organ injury in a large preclinical model. PLoS ONE 3 (3) : e1734. ScholarBank@NUS Repository. https://doi.org/10.1371/journal.pone.0001734 | Rights: | Attribution 4.0 International | Abstract: | Background: Current gene- and cell-based therapies have significant limitations which impede widespread clinical application. Taking diabetes mellitus as a paradigm, we have sought to overcome these limitations by ex vivo electrotransfer of a nonviral insulin expression vector into primary hepatocytes followed by immediate autologous reimplantation in a preclinical model of diabetes. Methods and Results: In a single 3-hour procedure, hepatocytes were isolated from a surgically resected liver wedge, electroporated with an insulin expression plasmid ex vivo and reimplanted intraparenchymally under ultrasonic guidance into the liver in each of 10 streptozotocin-induced diabetic Yorkshire pigs. The vector was comprised of a bifunctional, glucose-responsive promoter linked to human insulin cDNA. Ambient glucose concentrations approriately altered human insulin mRNA expression and C-peptide secretion within minutes in vitro and in vivo. Treated swine showed correction of hyperglycemia glucose intolerance, dyslipidemia and other metabolic abnormalities for ?47 weeks. Metabolic correction correlated significantly with the number of hepatocytes implanted. Importantly, we observed no hypoglycemia even under fasting conditions. Direct intrahepatic implantation of hepatocytes did not alter biochemical indices of liver function or induce abnormal hepatic lobular architecture. About 70% of implanted hepatocytes functionally engrafted, appeared histological normal, retained vector DNA and expressed human insulin for ?47 weeks. Based on structural tissue analyses and transcriptome data, we showed that early correction of diabetes attenuated and even prevented pathological changes in the eye, kidney, liver and aorta. Conclusions: We demonstrate that autologous hepatocytes can be efficiently, simple and safely modified by electroporation of a nonviral vector to express, process and secrete insulin durably. This strategy, which achieved significant and sustained therapeutic efficacy in a large preclinical model without adverse effects, warrants consideration for clinical development especially as it could have broader future applications for the treatment of other acquired and inherited diseases for which systemic reconstitution of a specific protein deficiency is critical. � 2008 Chen et al. | Source Title: | PLoS ONE | URI: | https://scholarbank.nus.edu.sg/handle/10635/161858 | ISSN: | 19326203 | DOI: | 10.1371/journal.pone.0001734 | Rights: | Attribution 4.0 International |
Appears in Collections: | Staff Publications Elements |
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