Please use this identifier to cite or link to this item: 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
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
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