Long-term survival of neonatal porcine islets in nonhuman primates by targeting costimulation pathways

Cardona, Kenneth; Korbutt, Gregory S.; Milas, Zvonimir L.; Lyon, James; Cano, Jose; Jiang, Wanhong; Bello-Laborn, Hameeda; Hacquoil, Brad V; Strobert, Elizabeth; Gangappa, Shivaprakash; Weber, Collin J.; Pearson, Thomas C.; Rajotte, Ray V.; Larsen, Christian

doi:10.1038/nm1375

Cited by 442 publications

(469 citation statements)

References 14 publications

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“…Very recently, the capacity of neonatal and adult porcine islets to restore normoglycemia in diabetic NHP was demonstrated (16,17). However, the immune suppression required to maintain the implants was either too toxic or included anti-CD40L, which cannot be used in human patients due to its thrombotic properties.…”

Section: Discussionmentioning

confidence: 99%

Embryonic pig pancreatic tissue for the treatment of diabetes in a nonhuman primate model

Hecht

Eventov‐Friedman

Rosen

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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Xenotransplantation of pig tissues has great potential to overcome the shortage of organ donors. One approach to address the vigorous immune rejection associated with xenotransplants is the use of embryonic precursor tissue, which induces and utilizes host vasculature upon its growth and development. Recently, we showed in mice that embryonic pig pancreatic tissue from embryonic day 42 (E42) exhibits optimal properties as a ␤ cell replacement therapy. We now demonstrate the proof of concept in 2 diabetic Cynomolgus monkeys, followed for 393 and 280 days, respectively. A marked reduction of exogenous insulin requirement was noted by the fourth month after transplantation, reaching complete independence from exogenous insulin during the fifth month after transplantation, with full physiological control of blood glucose levels. The porcine origin of insulin was documented by a radioimmunoassay specific for porcine C-peptide. Furthermore, the growing tissue was found to be predominantly vascularized with host blood vessels, thereby evading hyperacute or acute rejection, which could potentially be mediated by preexisting anti-pig antibodies. Durable graft protection was achieved, and most of the late complications could be attributed to the immunosuppressive protocol. While fine tuning of immune suppression, tissue dose, and implantation techniques are still required, our results demonstrate that porcine E-42 embryonic pancreatic tissue can normalize blood glucose levels in primates. Its long-term proliferative capacity, its revascularization by host endothelium, and its reduced immunogenicity, strongly suggest that this approach could offer an attractive replacement therapy for diabetes.immune-suppression ͉ rejection ͉ xeno-transplantation

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Section: Discussionmentioning

confidence: 99%

Embryonic pig pancreatic tissue for the treatment of diabetes in a nonhuman primate model

Hecht

Eventov‐Friedman

Rosen

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

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“…Pigs seem to be the best candidates for this purpose: they are readily available, produce a large progeny, and regulate blood glucose levels similarly to humans. In addition, porcine neonatal islet cell clusters (Cardona et al 2006;MacKenzie et al 2003) and islets from adult pigs (Hering et al 2006) are an interesting source of insulin-producing tissue for transplantation purposes. In this regard, there is an urgent need of genetic tools for the tissue-specific delivery of gene products to porcine pancreatic -cells.…”

Section: Introductionmentioning

confidence: 99%

Specific transgene expression in mouse pancreatic β-cells under the control of the porcine insulin promoter

Grzech¹,

Dahlhoff²,

Herbach³

et al. 2010

Molecular and Cellular Endocrinology

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The availability of regulatory sequences directing tissue-specific expression of transgenes in genetically modified mice and large animals is a prerequisite for the development of adequate models for human diseases. The rat insulin 2 gene (Ins2) promoter, widely used to achieve transgene expression in pancreatic -cells of mice, also directs expression to extrapancreatic tissues and performs poorly in isolated pancreatic islets of human, mouse, and pig. To evaluate whether the full 5' untranslated region (UTR) of the porcine insulin gene (INS) confers robust and specific expression in -cells we generated an expression cassette containing 1500 bp of the porcine INS 5' UTR and the 3' UTR of the bovine growth hormone gene (GH). The cassette was designed to allow easy exchange of the sequences to be expressed and easy removal of the vector backbone from the expression cassette. To evaluate the properties of the cassette, we initially cloned a cDNA encoding human betacellulin, a growth factor known to affect structural and functional parameters of -cells. After confirming the functionality and specificity of the construct in vitro, transgenic mouse lines were generated by pronuclear DNA microinjection. Using RT-PCR, immunohistochemistry and immunofluorescence, we show that transgenic mice expressed human betacellulin exclusively in -cells. Confirming the proposed insulinotropic effect of betacellulin, transgenic mice showed improved glucose tolerance. We conclude that the newly designed expression cassette containing 1.500 bp of the porcine insulin promoter 5' UTR confers robust and specific transgene expression to -cells in vitro and in transgenic mice.

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“…expand their beta cell mass following transplantation (23). Notably, porcine islets have been shown to maintain long-term function following intraportal transplantation into nonhuman primates with production of detectable porcine C-peptide and restoration of normoglycemia (24)(25)(26)(27)(28). In a limited number of case reports, xenotransplantation of porcine islets into diabetic humans has improved glycemic control (29).…”

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confidence: 99%

Islet amyloid deposition limits the viability of human islet grafts but not porcine islet grafts

Potter¹,

Abedini

Marek

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

159

160

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Islet transplantation is a promising treatment for diabetes but longterm success is limited by progressive graft loss. Aggregates of the beta cell peptide islet amyloid polypeptide (IAPP) promote beta cell apoptosis and rapid amyloid formation occurs in transplanted islets. Porcine islets are an attractive alternative islet source as they demonstrate long-term graft survival. We compared the capacity of transplanted human and porcine islets to form amyloid as an explanation for differences in graft survival. Human islets were transplanted into streptozotocin-diabetic immune-deficient mice. Amyloid deposition was detectable at 4 weeks posttransplantation and was associated with islet graft failure. More extensive amyloid deposition was observed after 8 weeks. By contrast, no amyloid was detected in transplanted neonatal or adult porcine islets that had maintained normoglycemia for up to 195 days. To determine whether differences in IAPP sequence between humans and pigs could explain differences in amyloid formation and transplant viability, we sequenced porcine IAPP. Porcine IAPP differs from the human sequence at 10 positions and includes substitutions predicted to reduce its amyloidogenicity. Synthetic porcine IAPP was considerably less amyloidogenic than human IAPP as determined by transmission electron microscopy, circular dichroism, and thioflavin T binding. Viability assays indicated that porcine IAPP is significantly less toxic to INS-1 beta cells than human IAPP. Our findings demonstrate that species differences in IAPP sequence can explain the lack of amyloid formation and improved survival of transplanted porcine islets. These data highlight the potential of porcine islet transplantation as a therapeutic approach for human diabetes.

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Long-term survival of neonatal porcine islets in nonhuman primates by targeting costimulation pathways

Cited by 442 publications

References 14 publications

Embryonic pig pancreatic tissue for the treatment of diabetes in a nonhuman primate model

Embryonic pig pancreatic tissue for the treatment of diabetes in a nonhuman primate model

Specific transgene expression in mouse pancreatic β-cells under the control of the porcine insulin promoter

Islet amyloid deposition limits the viability of human islet grafts but not porcine islet grafts

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