Diabetes mellitus is a growing problem worldwide. In the US it affects over 18 million people and results in annual health care costs exceeding $130 billion. Insulin therapy of Type 1 diabetes, and in advanced cases of Type 2 diabetes, does not prevent serious long-term complications including neuropathy, vascular disease, retinopathy and renal failure. Transplantation of pancreatic islets to restore insulin production offers significant promise. However, the supply of donor pancreata falls far short of meeting the medical need. New sources of insulin producing cells will be required to realize the full potential of cell therapy for diabetes. We propose to generate pancreatic beta lineage cells by in vitro differentiation of stem cells isolated from amniotic fluid. These """"""""AFS cells"""""""" are capable of both extensive expansion and differentiation into derivatives of all three embryonic germ layers. Our Preliminary Studies showed that mouse AFS cells can yield insulin producing cells and islet- like cell clusters (""""""""neo-islets""""""""), promoted by expression of the pancreatic transcription factor PDX-1. We now propose to produce neo-islets from human and nonhuman primate (NHP) AFS cells. To efficiently generate insulin producing cells, we will optimize delivery of a plasmid vector to express PDX-1, and will systematically test growth factors and substrates shown previously to promote pancreatic beta cell differentiation. The resulting neo-islets will be compared with authentic human and NHP pancreatic islets using tests developed for clinical transplantation. To assess their ability to restore control of glucose metabolism and production of insulin and C-peptide, neo-islets will be implanted in immunodeficient mice made diabetic with streptozotocin (STZ). Transplantation in STZ-treated NHP will assess the function of neo-islets in a model physiologically more similar to humans. NHP AFS cell lines will be derived after amniocentesis of pregnant mothers. These stem cells will be used to generate neo-islets for autologous transplantation into the corresponding offspring. The same donor cells will be compared in allogeneic recipients using clinically relevant immunosuppression regimens. Successful development of an abundant source of transplantable insulin producing cells potentially would have a profound impact on the treatment of a major public health problem.

Public Health Relevance

Transplantation of insulin producing cells of the pancreas may prevent the severe long-term complications faced by millions of diabetes patients, but is limited by the extreme scarcity of donated organs. Stem cells have been isolated from a novel source, namely, amniotic fluid collected for prenatal genetic testing. The proposed program will utilize amniotic stem cells to generate an abundant supply of insulin producing cells which will be tested for their ability to restore control of blood sugar levels in preclinical models of diabetes.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
3R01EB008009-01S1
Application #
7630939
Study Section
Special Emphasis Panel (ZEB1-OSR-C (O1))
Program Officer
Hunziker, Rosemarie
Project Start
2008-08-01
Project End
2009-07-31
Budget Start
2008-08-01
Budget End
2009-07-31
Support Year
1
Fiscal Year
2008
Total Cost
$111,000
Indirect Cost
Name
Wake Forest University Health Sciences
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
937727907
City
Winston-Salem
State
NC
Country
United States
Zip Code
27157
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Faleo, Gaetano; Berggren, Per-Olof; Pileggi, Antonello (2014) Intravital imaging of cytotoxic T lymphocytes. Methods Mol Biol 1186:121-9
Zhou, Yu; Mack, David L; Williams, J Koudy et al. (2013) Genetic modification of primate amniotic fluid-derived stem cells produces pancreatic progenitor cells in vitro. Cells Tissues Organs 197:269-82
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Pileggi, Antonello; Ricordi, Camillo (2013) A new home for pancreatic islet transplants: the bone marrow. Diabetes 62:3333-5
Pileggi, Antonello; Xu, Xiumin; Tan, Jianming et al. (2013) Mesenchymal stromal (stem) cells to improve solid organ transplant outcome: lessons from the initial clinical trials. Curr Opin Organ Transplant 18:672-81
Pileggi, Antonello (2012) Mesenchymal stem cells for the treatment of diabetes. Diabetes 61:1355-6
Skardal, Aleksander; Mack, David; Kapetanovic, Edi et al. (2012) Bioprinted amniotic fluid-derived stem cells accelerate healing of large skin wounds. Stem Cells Transl Med 1:792-802

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