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Am J Transl Res 2013;5(2):184-199

Original Article
Genetically reprogrammed, liver-derived insulin-producing cells are
glucose-responsive, but susceptible to autoimmune destruction in
settings of murine model of type 1 diabetes

Dong-Qi Tang, Lu Shun, Vijay Koya, Yuping Sun, Qiwei Wang, Hai Wang, Shi-Wu Li, Yu Sun, Daniel L Purich,
Clare Zhang, Barbara Han-sen, Keping Qian, Mark Atkinson, M Ian Phillips, Li-Jun Yang

Department of Pathology, Immunology, and Laboratory Medicine, Department of Biochemistry & Molecular
Biology, University of Florida Col-lege of Medicine, Gainesville, Florida 32610, USA; Obesity, Diabetes and Aging
Research Center and Departments of Internal Medicine and Pediatrics, College of Medicine, University of South
Florida, Tampa, FL 33612, USA; Center for Rare Disease Therapies, Keck Graduate Insti-tute, Claremont,
California, 91711, USA. *The first two authors contributed to this work equally.

Received February 20, 2013; Accepted March 8, 2013; Epub March 28, 2013; Published April 8, 2013

Abstract: Many previous studies demonstrate that hepatocytes can be reprogrammed into insulin-producing
cells (IPCs) utilizing viral vec-tor-mediated delivery of pancreatic transcription factors (PTFs). However, whether
these liver-derived IPCs are susceptible to autoimmune attack in animal models of type 1 diabetes remains
unclear, in part due to the immunogenicity of the viral vectors used to introduce PTF genes. Adeno-associated
virus serotype 2 vector-expressing Pdx1-VP16 (Pdx1) and Ngn3 were prepared and injected into the portal vein of
streptozotocin (Stz)/diabetic NOD/SCID mice. The presence of glucose-responsive liver-IPCs and their
susceptibility to anti-beta cell autoim-munity were assessed by blood glucose levels, insulin content, IPC cell
distribution, and intraperitoneal glucose tolerance test following subtotal pancreatectomy (Px) and passive
transfer of diabetogenic splenocytes isolated from diabetic female NOD mice. A combination of two PTF genes
(Pdx1/Ngn3) effectively reprogrammed liver cells into glucose-responsive IPCs. These IPCs corrected
hyperglycemia in Stz/diabetic NOD/SCID mice and maintained normoglycemia following subtotal Px, indicating
that liver-derived IPCs could maintain glucose homeostasis. Importantly, we also demonstrated that the glucose-
responsive liver–derived IPCs were susceptible to autoimmune destruction by diabetogenic splenocytes, as
indicated by progressive elevation in blood glucose levels as well as mixed T-, and B-lymphocytic infiltrates
surrounding liver-IPCs 2~3 weeks following transferring of diabetogenic splenocytes into NOD/SCID mice, and
confirmed by immunohistochemical studies. In conclusion, genetically reprogrammed liver-IPCs, like pancreatic
islet beta-cells, are susceptible to autoimmune attack, suggesting that for cell-replacement therapy of treating
type 1 diabetes, beta-cell surrogates may require concomitant immunotherapy to avoid autoimmune destruction

Keywords: Reprogram, hepatocytes, insulin-producing beta cells, autoimmunity, type 1 diabetes, gene therapy

Address correspondence to: Dr. Li-Jun Yang, Department of Pathology, Immunology, and Laboratory Medicine,
University of Florida College of Medicine, 1600 SW Archer Road, P.O. Box 100275, Gainesville, FL 32610, U.S.A.
Phone: 352-392-0005; E-mail: yanglj@pathology.ufl.edu