Constitutive and regulated expression of processed insulin following in vivo hepatic gene transfer

Auricchio, Alberto; Gp, Gao; Qc, Yu; Raper, Steven E.; Rivera, Victor M.; Clackson, Tim; Wilson, James M.

doi:10.1038/sj.gt.3301746

Cited by 79 publications

(49 citation statements)

References 42 publications

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“…Cleavage of modified proinsulin is likely to occur in the trans-Golgi network where furin is localized 13 and where transgenic insulin expressed in hepatocytes has been located. 14 A third modification was introduced because the HisB10Asp variant has four-to five-fold higher biological potency than native insulin, possibly due to higher avidity of receptor binding 15 and more rapid secretion via the constitutive pathway. …”

Section: Discussionmentioning

confidence: 99%

Plasmid-electroporated primary hepatocytes acquire quasi-physiological secretion of human insulin and restore euglycemia in diabetic mice

et al. 2005

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We describe the durable correction of streptozotocin-induced murine diabetes by in vivo implantation of primary mouse hepatocytes electroporated ex vivo with a human proinsulin cDNA plasmid construct controlled by glucose and zinc regulatory elements. Transfected hepatocytes increased insulin transgene transcription and secretion within 10-20 min of exposure to 25 mM glucose or 60 mM zinc. Insulin release did not occur from secretory granules. Electroporated Rosa26 hepatocytes (B8 Â 10 5 viable cells) were implanted in C57BL/6J diabetic mice in one of three sites: unresected liver, regenerating liver or mesentery. Control diabetic mice were implanted with untransfected hepatocytes. At 30 days after implantation, 8/15 control mice were alive, while 19/19 treated mice were alive. The ratio of body weight on day 30/nadir body weight was significantly higher for all treated groups compared with controls. All eight surviving control mice were hyperglycemic 30 days postimplantation, while 16/19 treated diabetic mice remained normoglycemic. Treated mice had lower mean glucose values (Pp0.001) without fasting hypoglycemia and better glucose tolerance (Pp0.0003) than untreated controls. All (6/6) diabetic mice implanted in regenerating liver and 71% (5/7) implanted in unresected liver were alive 77 days after implantation. Engrafted hepatocytes were identified, mainly around central veins, by staining for b-galactosidase activity and with anti-human insulin antibody. Gene Therapy (2005) 12, 655-667.

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Section: Discussionmentioning

confidence: 99%

Plasmid-electroporated primary hepatocytes acquire quasi-physiological secretion of human insulin and restore euglycemia in diabetic mice

et al. 2005

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“…Removal of the nonessential E3 region further increased the cloning capacity to 8 kb. 21 There are numerous examples of the successful application of Ad in gene therapy of animal models of various autoimmune diseases including diabetes, 22 rheumatoid arthritis (RA), 23 multiple sclerosis (MS), 24 25 Sjogren's syndrome, 26 autoimmune myocarditis, 27 and autoimmune uveitis. 28 The routes of Ad delivery in these studies include i.v., 25 intra-articular (i.a.)…”

Section: Dna Complexesmentioning

confidence: 99%

“…180 Other in vivo studies have also examined systems in muscle. 10,22 Transcriptionally regulated systems are characterized by a significant time lag (approximately 6 h) between induction of gene expression and protein synthesis. In comparison, secretion of endogenous insulin from b islet cells occurs within approximately 20 min after glucose uptake.…”

Section: Targeting Immune Cellsmentioning

confidence: 99%

Vectors for the treatment of autoimmune disease

Gould

Favorov

2003

Gene Ther

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“…Rivera et al demonstrated in different cells with a constitutive pathway that proteins can be accumulated as aggregates in the endoplasmic reticulum and secreted by the stimulation of a synthetic small-molecule drug that induces protein disaggregation (105). Thus, in vivo experiments using this strategy have already proven this principle that insulin secretion in hepatocytes can be triggered by rapamycin (61). An emerging approach that holds promise is the transdifferentiation of hepatocytes into insulin-secreting cells that are capable of storing and secreting insulin in a very similar way like b-cells.…”

Section: Regulation Of Insulin Secretion In Transduced Hepatocytementioning

confidence: 99%