Sertoli Cells Engineered to Express Insulin to Lower Blood Glucose in Diabetic Mice

Long-term survival of allo- and xenotransplanted immune-privileged Sertoli cells (SCs) is well documented suggesting that SCs can be used to deliver foreign proteins for cell-based gene therapy. The aim of this study was to use a lentivirus carrying proinsulin cDNA to achieve stable expression and lowering of blood glucose levels (BGLs). A SC line transduced with the lentivirus (MSC-LV-mI) maintained stable insulin expression in vitro. These MSC-LV-mI cells were transplanted and grafts were analyzed for cell survival, continued proinsulin mRNA, and insulin protein expression. All grafts contained MSC-LV-mI cells that expressed proinsulin mRNA and insulin protein. Transplantation of MSC-LV-mI cells into diabetic mice significantly lowered BGLs for 4 days after transplantation. Interestingly, in three transplanted SCID mice and one transplanted BALB/c mouse, the BGLs again significantly lowered by day 50 and 70, respectively. This is the first time SC transduced with a lentiviral vector was able to stably express insulin and lower BGLs. In conclusion, a SC line can be modified to stably express therapeutic proteins (e.g., insulin), thus taking us one step further in the use of SCs as an immune-privileged vehicle for cell-based gene therapy.

[1]  J. Dufour,et al.  Xenotransplanted Pig Sertoli Cells Inhibit Both the Alternative and Classical Pathways of Complement-Mediated Cell Lysis While Pig Islets Are Killed , 2016, Cell transplantation.

[2]  J. Dufour,et al.  Sertoli cells--immunological sentinels of spermatogenesis. , 2014, Seminars in cell & developmental biology.

[3]  J. Dufour,et al.  Sustained Expression of Insulin by a Genetically Engineered Sertoli Cell Line after Allotransplantation in Diabetic BALB/c Mice1 , 2014, Biology of reproduction.

[4]  L. Spain,et al.  Development of standardized insulin treatment protocols for spontaneous rodent models of type 1 diabetes. , 2012, Comparative medicine.

[5]  J. Dufour,et al.  Genetically engineered immune privileged Sertoli cells , 2012, Spermatogenesis.

[6]  J. Dufour,et al.  The Blood-Testis and Blood-Epididymis Barriers Are More than Just Their Tight Junctions1 , 2011, Biology of reproduction.

[7]  J. Dufour,et al.  Delivery of a Therapeutic Protein by Immune-Privileged Sertoli Cells , 2010, Cell transplantation.

[8]  R. Herzog,et al.  Progress and prospects: immune responses to viral vectors , 2010, Gene Therapy.

[9]  J. Dufour,et al.  Immunoprotective sertoli cells: making allogeneic and xenogeneic transplantation feasible. , 2010, Reproduction.

[10]  D. White,et al.  Diabetic rats and mice are resistant to porcine and human insulin: flawed experimental models for testing islet xenografts , 2009, Xenotransplantation.

[11]  P. van Endert,et al.  Therapy of experimental type 1 diabetes by isolated Sertoli cell xenografts alone , 2009, The Journal of experimental medicine.

[12]  James M. Wilson,et al.  Activation of Transgene-specific T Cells Following Lentivirus-mediated Gene Delivery to Mouse Lung , 2009, Molecular Therapy.

[13]  David A. Williams.  Gene therapy continues to mature and to face challenges. , 2009, Molecular therapy : the journal of the American Society of Gene Therapy.

[14]  Frederic D Bushman,et al.  Analysis of lentiviral vector integration in HIV+ study subjects receiving autologous infusions of gene modified CD4+ T cells. , 2009, Molecular therapy : the journal of the American Society of Gene Therapy.

[15]  D. Parsons,et al.  Gene delivery to airway epithelial cells in vivo: a direct comparison of apical and basolateral transduction strategies using pseudotyped lentivirus vectors , 2007, The journal of gene medicine.

[16]  Clelia Di Serio,et al.  Hematopoietic stem cell gene transfer in a tumor-prone mouse model uncovers low genotoxicity of lentiviral vector integration , 2006, Nature Biotechnology.

[17]  Takuji Igarashi,et al.  Suitability of allogeneic sertoli cells for ex vivo gene delivery in the injured spinal cord , 2006, Experimental Neurology.

[18]  G. Korbutt,et al.  Genetically engineered Sertoli cells are able to survive allogeneic transplantation , 2004, Gene Therapy.

[19]  Adam Bagg,et al.  Fatal systemic inflammatory response syndrome in a ornithine transcarbamylase deficient patient following adenoviral gene transfer. , 2003, Molecular genetics and metabolism.

[20]  G. Korbutt,et al.  Immunoprotection of rat islet xenografts by cotransplantation with sertoli cells and a single injection of antilymphocyte serum1 , 2003, Transplantation.

[21]  Mark A. Kay,et al.  Progress and problems with the use of viral vectors for gene therapy , 2003, Nature Reviews Genetics.

[22]  G. Korbutt,et al.  Long-term survival of intratesticular porcine islets in nonimmunosuppressed beagles , 2003, Transplantation.

[23]  M. Kay,et al.  Modified HIV-1 based lentiviral vectors have an effect on viral transduction efficiency and gene expression in vitro and in vivo. , 2001, Molecular therapy : the journal of the American Society of Gene Therapy.

[24]  B. Harder,et al.  Lentivirus vectors encoding both central polypurine tract and posttranscriptional regulatory element provide enhanced transduction and transgene expression. , 2001, Human gene therapy.

[25]  G. Korbutt,et al.  Testicular sertoli cells protect islet beta-cells from autoimmune destruction in NOD mice by a transforming growth factor-beta1-dependent mechanism. , 2000, Diabetes.

[26]  M. Lusky,et al.  Gene therapy with recombinant adenovirus vectors: evaluation of the host immune response. , 1997, Immunology letters.

[27]  G. Korbutt,et al.  Cotransplantation of Allogeneic Islets With Allogeneic Testicular Cell Aggregates Allows Long-Term Graft Survival Without Systemic Immunosuppression , 1997, Diabetes.

[28]  K. Tung,et al.  Autoantigenic germ cells exist outside the blood testis barrier. , 1988, Journal of immunology.

[29]  N. Mann,et al.  Human insulin and porcine insulin in the treatment of diabetic children: comparison of metabolic control and insulin antibody production. , 1983, British medical journal.

[30]  L. Romrell,et al.  Appearance of cell surface auto- and isoantigens during spermatogenesis in the rabbit. , 1977, Developmental biology.

[31]  S. Gray,et al.  Viral expression cassette elements to enhance transgene target specificity and expression in gene therapy. , 2015, Discovery medicine.

[32]  G. Korbutt,et al.  Nondividing, Postpubertal Rat Sertoli Cells Resumed Proliferation after Transplantation1 , 2014, Biology of reproduction.

[33]  M. Karaca,et al.  Transgenic expression of human INS gene in Ins1/Ins2 double knockout mice leads to insulin underproduction and diabetes in some male mice. , 2007, Frontiers in bioscience : a journal and virtual library.

[34]  J. Altomonte,et al.  Basal insulin gene expression significantly improves conventional insulin therapy in type 1 diabetic rats. , 2002, Diabetes.