Transplantation of Genetically Engineered Insulin-Producing Hepatocytes Into Immunoincompetent Mice

Tuch, B E.; Tabiin, Muhammad T; Casamento, Frances M.; Simpson, Ann M.; Marshall, G M

doi:10.1016/s0041-1345(97)01361-4

Cited by 8 publications

(8 citation statements)

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“…6 The ability of the transplanted HEP G2ins/g cells to secrete (pro)insulin in response to glucose was impaired, with the cells having no immediate effect on the blood glucose levels of the diabetic recipient mice. 24 This was different from the results we obtained with transplanted HUH7-ins cells ( Figure 10). Our results with the HUH7-ins cells support the conclusion of other researchers, [7][8][9][10][11][12][13] namely that liver cells, or at least their precursors, 9 can be genetically altered to act as b cells, and replace those which are lost in type I diabetes.…”

Section: Discussioncontrasting

confidence: 94%

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Function of a genetically modified human liver cell line that stores, processes and secretes insulin

Tuch

Szymańska

et al. 2003

Gene Ther

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An alternative approach to the treatment of type I diabetes is the use of genetically altered neoplastic liver cells to synthesize, store and secrete insulin. To try and achieve this goal we modified a human liver cell line, HUH7, by transfecting it with human insulin cDNA under the control of the cytomegalovirus promoter. The HUH7-ins cells created were able to synthesize insulin in a similar manner to that which occurs in pancreatic b cells. They secreted insulin in a regulated manner in response to glucose, calcium and theophylline, the dose-response curve for glucose being near-physiological. Perifusion studies showed that secretion was rapid and tightly controlled. Removal of calcium resulted in loss of glucose stimulation while addition of brefeldin A resulted in a 30% diminution of effect, indicating that constitutive release of insulin occurred to a small extent. Insulin was stored in granules within the cytoplasm. When transplanted into diabetic immunoincompetent mice, the cells synthesized, processed, stored and secreted diarginyl insulin in a rapid regulated manner in response to glucose.Constitutive release of insulin also occurred and was greater than regulated secretion. Blood glucose levels of the mice were normalized but ultimately became subnormal due to continued proliferation of cells. Examination of the HUH7-ins cells as well as the parent cell line for b cell transcription factors showed the presence of NeuroD but not PDX-1. PC1 and PC2 were also present in both cell types. Thus, the parent HUH7 cell line possessed a number of endocrine pancreatic features that reflect the common endodermal ancestry of liver and pancreas, perhaps as a result of ontogenetic regression of the neoplastic liver cell from which the line was derived. Introduction of the insulin gene under the control of the CMV promoter induced changes in these cells to make them function to some extent like pancreatic b cells. Our results support the view that neoplastic liver cells can be induced to become substitute pancreatic b cells and become a therapy for the treatment of type I diabetes.

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Section: Discussioncontrasting

confidence: 94%

“…11 The HEP G2ins/g cells do secrete (pro)insulin rapidly in response to glucose in vitro but the cells are unstable when transplanted. 24 Secretion of the singlechain insulin analog from rats does occur when glucose is administered, but is delayed with the peak effect at 4 hours, presumably because control is at a translational level.…”

Section: Discussionmentioning

confidence: 99%

Function of a genetically modified human liver cell line that stores, processes and secretes insulin

Tuch

Szymańska

et al. 2003

Gene Ther

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“…[26][27][28][29] However, transfer of these regulatory mechanisms to in vivo models has been difficult. 5 In contrast, data obtained from in vitro studies using our transgene, appear to accurately reflect function in vivo. 10 We are proceeding to describe the secretory dynamics of our system, and anticipate that refinements to our system will reduce abnormal glycemic excursions.…”

Section: Used Continuousmentioning

confidence: 83%

“…[2][3][4] However, attempts to regulate transgenic insulin production have proven inadequate. 5,6 Consequently, secretion of transgenic insulin has been either insufficient to normalize blood glucose, 4,[6][7][8][9] or has produced lethal hypoglycemia. 3,4,7 We have designed a system of insulin gene therapy that utilizes transcription to regulate hepatic production of transgenic insulin.…”

Section: Introductionmentioning

confidence: 99%

Regulated hepatic insulin gene therapy of STZ-diabetic rats

2000

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Effective and safe insulin gene therapy will require regulation of transgenic insulin secretion. We have created a livertargeted insulin transgene by engineering glucose responsive elements into a hepatic promoter containing an inhibitory insulin response sequence. In this work, we demonstrate application of this transgene for the treatment of diabetes mellitus in vivo, by administering a recombinant adenovirus vector, Ad/(GlRE) 3 BP-1 2xfur, to rats made diabetic with streptozotocin. We verified hepatic expression of transgenic insulin by RT-PCR, and confirmed glucose responsive stimulation of transgenic insulin secretion in vivo by serum RIA. Following a portal system injection of either Ad/(GlRE) 3 BP-1 2xfur, or an empty adenoviral vector, animals made diabetic with either low (120 mg/kg), or high (290 mg/kg) dose streptozotocin (STZ) were monitored for changes in body weight, and blood glucose. Without subcutaneous insulin injections, blood glucose values of sham-

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“…In the second approach, hepatoma-derived cell lines have been modified for glucose-inducible insulin secretion in vitro. However, these engineered hepatoma cells either failed to be regulated in vivo [11] or expressed insulin at levels that were deemed too low for testing in vivo [12]. Notwithstanding these problems, hepatocyte-derived in vivo bioimplants are serious candidates for cellular therapy of diabetes mellitus, not only because liver and b cells have shared physiological attributes, but also because primary hepatocytes can be genetically modified ex vivo to acquire desired phenotypic features that provide therapeutic effects in vivo after reimplantation [13,14].…”

Section: Introductionmentioning

confidence: 99%

Human liver‐derived cells stably modified for regulated proinsulin secretion function as bioimplants in vivo

Chen

Patil

Lok

et al. 2002

The Journal of Gene Medicine

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Transplantation of Genetically Engineered Insulin-Producing Hepatocytes Into Immunoincompetent Mice

Cited by 8 publications

References 1 publication

Function of a genetically modified human liver cell line that stores, processes and secretes insulin

Function of a genetically modified human liver cell line that stores, processes and secretes insulin

Regulated hepatic insulin gene therapy of STZ-diabetic rats

Human liver‐derived cells stably modified for regulated proinsulin secretion function as bioimplants in vivo

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