Lentiviral transduction of rat Sertoli cells as a means to modify gene expression

Nicholls, Peter K.; Stanton, Peter G.; Rainczuk, Katarzyna; Qian, Hongwei; Gregorevic, Paul; Harrison, Craig A.

doi:10.4161/spmg.22516

Cited by 2 publications

(2 citation statements)

References 36 publications

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“…The ability of immune-privileged Sertoli cells to survive across immunological barriers (i.e., allo-or xenotransplantation in rodents and humans without immunosuppressive drugs [5,7,12,25]) make them an ideal vehicle for cell-based gene therapy. Viral (adenovirus, retrovirus, and lentivirus) vectors have been used successfully to transduce Sertoli cells [34][35][36][37]. For instance, in vivo transduction of pubertal ''nondividing'' Sertoli cells with an adenoviral vector carrying the full-length mouse Steel gene partially restored spermatogenesis in infertile Steel/Steel dickie (Sl/Sl d ) mutant mouse testes at 6-10 wk after virus injection.…”

Section: Discussionmentioning

confidence: 98%

Sustained Expression of Insulin by a Genetically Engineered Sertoli Cell Line after Allotransplantation in Diabetic BALB/c Mice1

Kaur

Thompson

Pasham

et al. 2014

Biology of Reproduction

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Immune-privileged Sertoli cells (SCs) exhibit long-term survival after allotransplantation or xenotransplantation, suggesting they can be used as a vehicle for cell-based gene therapy. Previously, we demonstrated that SCs engineered to secrete insulin by using an adenoviral vector normalized blood glucose levels in diabetic mice. However, the expression of insulin was transient, and the use of immunocompromised mice did not address the question of whether SCs can stably express insulin in immunocompetent animals. Thus, the objective of the current study was to use a lentiviral vector to achieve stable expression of insulin in SCs and test the ability of these cells to survive after allotransplantation. A mouse SC line transduced with a recombinant lentiviral vector containing furin-modified human proinsulin cDNA (MSC-EhI-Zs) maintained stable insulin expression in vitro. Allotransplantation of MSC-EhI-Zs cells into diabetic BALB/c mice demonstrated 88% and 75% graft survival rates at 20 and 50 days post-transplantation, respectively. Transplanted MSC-EhI-Zs cells continued to produce insulin mRNA throughout the study (i.e., 50 days); however, insulin protein was detected only in patches of cells within the grafts. Consistent with low insulin protein detection, there was no significant change in blood glucose levels in the transplant recipients. Nevertheless, MSC-EhI-Zs cells isolated from the grafts continued to express insulin protein in culture. Collectively, this demonstrates that MSC-EhI-Zs cells stably expressed insulin and survived allotransplantation without immunosuppression. This further strengthens the use of SCs as targets for cell-based gene therapy for the treatment of numerous chronic diseases, especially those that require basal protein expression.

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Section: Discussionmentioning

confidence: 98%

Sustained Expression of Insulin by a Genetically Engineered Sertoli Cell Line after Allotransplantation in Diabetic BALB/c Mice1

Kaur

Thompson

Pasham

et al. 2014

Biology of Reproduction

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“…Ted and Valina Dawson from The Johns Hopkins University School of Medicine (Baltimore, MD); it was prepared (24) and lentiviral particles were generated as described previously (25).…”

Section: Construction and Generation Of Lentiviral Vectorsmentioning

confidence: 99%

Deficiency in Apoptosis-Inducing Factor Recapitulates Chronic Kidney Disease via Aberrant Mitochondrial Homeostasis

et al. 2016

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Apoptosis-inducing factor (AIF) is a mitochondrial flavoprotein with dual roles in redox signaling and programmed cell death. Deficiency in AIF is known to result in defective oxidative phosphorylation (OXPHOS), via loss of complex I activity and assembly in other tissues. Because the kidney relies on OXPHOS for metabolic homeostasis, we hypothesized that a decrease in AIF would result in chronic kidney disease (CKD). Here, we report that partial knockdown of Aif in mice recapitulates many features of CKD, in association with a compensatory increase in the mitochondrial ATP pool via a shift toward mitochondrial fusion, excess mitochondrial reactive oxygen species production, and Nox4 upregulation. However, despite a 50% lower AIF protein content in the kidney cortex, there was no loss of complex I activity or assembly. When diabetes was superimposed onto Aif knockdown, there were extensive changes in mitochondrial function and networking, which augmented the renal lesion. Studies in patients with diabetic nephropathy showed a decrease in AIF within the renal tubular compartment and lower AIFM1 renal cortical gene expression, which correlated with declining glomerular filtration rate. Lentiviral overexpression of Aif1m rescued glucose-induced disruption of mitochondrial respiration in human primary proximal tubule cells. These studies demonstrate that AIF deficiency is a risk factor for the development of diabetic kidney disease.

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Lentiviral transduction of rat Sertoli cells as a means to modify gene expression

Cited by 2 publications

References 36 publications

Sustained Expression of Insulin by a Genetically Engineered Sertoli Cell Line after Allotransplantation in Diabetic BALB/c Mice1

Sustained Expression of Insulin by a Genetically Engineered Sertoli Cell Line after Allotransplantation in Diabetic BALB/c Mice1

Deficiency in Apoptosis-Inducing Factor Recapitulates Chronic Kidney Disease via Aberrant Mitochondrial Homeostasis

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