Gene therapy for diabetes: strategies for β-cell modification and replacement

Levine, Fred

doi:10.1002/(sici)1099-0895(199712)13:4<209::aid-dmr198>3.0.co;2-n

Cited by 25 publications

(11 citation statements)

References 137 publications

(181 reference statements)

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“…Genetic engineering may potentially provide transplantable b cells with resistance to cytokine-media ted damage and in particular to cytokine-media ted apoptosis (Levine, 1997;Efrat, 1998;Iwahashi et al, 1998). For instance, insulinoma cells have been engineered to permanently express manganese superoxide dismutase, thus preventing IL-1b -induced b cell cytotoxicity via downregulation of iNOS transcription (Hohmeier et al, 1998).…”

Section: Discussionmentioning

confidence: 99%

“…These advantages facilitate evaluating the ex vivo efficiency of ribozymes, since one can avoid the variability and limitations associated with primary cell cultures. We chose to work primarily with the b TC-3 cell line because of its wide use and because its phenotype has been extensively characterized (Levine, 1997). Moreover, studies of this cell line include the demonstration, obviously relevant to our study, that b TC-3 cells maintain an active caspase 3 response in Fas-mediated apoptosis (Yamada et al, 1999).…”

Section: Discussionmentioning

confidence: 99%

“…In this category, and relevant to the anti-Fas ribozyme presented here, experimental islet cell transplantation is a logical testing ground for ribozyme-med iated manipulation of gene expression for therapeutic purposes. The continuous developm ent and increasing availability of new viral vectors for the delivery of genes into live primary cells, and in particular into pancreatic islets cells (Levine, 1997;Leibowitz et al, 1999;Konkal-Matt et al, 1999), should allow the testing of ribozymes in in vivo experimental models of islet cell transplantation.…”

Section: Color Platementioning

confidence: 99%

“…In particular, proapoptotic genes, such as those encoding the inducible nitric oxide synthase (iNOS) and Fas, are activated by proinflamm atory cytokines and significantly impair graft survival by inducing apoptosis of transplanted islet cells (Corbett and McDaniel, 1992;Stassi et al, 1995;Eizirik et al, 1996;Mandrup-Poul sen, 1996;Suarez-Pinzon et al, 1999). Genetic engineering could improve the outcom e of islet transplantation if transplantable islets or artificial b cells could be rendered less susceptible to apoptosis or other noxae (Levine, 1997;Efrat, 1998;Iwahashi et al, 1998). In experimental models, this goal has been achieved by introducing anti-apoptotic genes into b -cells (Hotta et al, 1998;Rabinovitch et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

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Inhibition of Fas-Mediated Apoptosis in Mouse Insulinoma betaTC-3 Cells via an Anti-Fas Ribozyme

Klein

Ricordi²,

Pugliese³

et al. 2000

Human Gene Therapy

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In this study we have designed and constructed an anti-Fas ribozyme and show that it can specifically cleave the Fas mRNA in vitro. Moreover, to test its efficacy ex vivo, we transfected the anti-Fas ribozyme into betaTC-3 insulinoma cells, using a RNA polymerase III promoter to drive its expression. Like pancreatic beta cells, betaTC-3 cells do not constitutively express Fas, but Fas expression can be induced with IL-1 and IFN-gamma. Transfected cells expressed an average of 5000 copies of anti-Fas ribozyme transcript per cell as assessed by reverse transcriptase-real-time PCR. After IL-1/IFN-gamma treatment, betaTC-3 cells transfected with the anti-Fas ribozyme expressed 80% less Fas compared with mock-transfected cells. In addition, the anti-Fas ribozyme also inhibited Fas expression in NIT-1 insulinoma cells and in primary cultures of dispersed pancreatic islet cells. Inhibition of de novo Fas expression in betaTC-3 cells expressing the anti-Fas ribozyme correlated with resistance to Fas-mediated apoptosis as determined by the number of cells exhibiting caspase 3 proteolytic activity. Hence, we have engineered a ribozyme capable of preventing Fas expression in the betaTC-3 pancreatic insulinoma cell line and conferring resistance to Fas-mediated apoptosis. We suggest that ribozymes may be potentially useful to engineer resistance to apoptosis in transplantable beta cells, a feature that may significantly improve the survival of islet cell grafts.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Color Platementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Inhibition of Fas-Mediated Apoptosis in Mouse Insulinoma betaTC-3 Cells via an Anti-Fas Ribozyme

Klein

Ricordi²,

Pugliese³

et al. 2000

Human Gene Therapy

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“…The development of in vitro culture techniques to generate large populations of beta cells from human cadaveric islets is an attractive approach to increasing the number of cells available for transplantation [63][64][65]. In vitro culture systems are advantageous for studying islet cell proliferation because the specific effects of particular trophic factors on growth and function can be tested in a controlled manner, individually or in defined combinations.…”

Section: Beta Cells From Beta Cells: Ex Vivo Expansion Of Primary Celmentioning

confidence: 99%

Diabetes Mellitus-Cell Transplantation and Gene Therapy Approaches.

Halvorsen¹,

Levine²

2001

CMM

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Diabetes mellitus affects millions of people in the United States and worldwide. It has become clear over the past decade that the chronic complications of diabetes result from lack of proper blood glucose concentration regulation, and particularly the toxic effects of chronic hyperglycemia on organs and tissues. Pancreas transplants can cure insulin-dependent diabetes mellitus (IDDM). Furthermore, recent advances in pancreatic islet isolation and immunosuppressive regimens have resulted in dramatic improvements in the survival and function of islet allografts. Therefore, islet replacement strategies are becoming increasingly attractive options for patients at risk for severe diabetic complications. A major limitation of these approaches is the small number of organs available for transplantation or islet isolation. Thus, an important next step in developing curative treatments for type I diabetes will be the generation of a replenishable source of glucose-responsive, insulin-secreting cells that can be used for beta cell replacement. This review focuses on approaches to developing robust and widely applicable beta-cell replacement strategies with an emphasis on manipulating beta-cell growth and differentiation by genetic engineering.

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