Kidney-specific transposon-mediated gene transfer in vivo.
Methods enabling kidney-specific gene transfer in adult mice are needed to develop new therapies for kidney disease. We attempted kidney-specific gene transfer following hydrodynamic tail vein injection using the kidney-specific podocin and gamma-glutamyl transferase promoters, but found expression primarily in the liver. In order to achieve kidney-specific transgene expression, we tested direct hydrodynamic injection of a DNA solution into the renal pelvis and found that luciferase expression was strong in the kidney and absent from extra-renal tissues. We observed heterogeneous, low-level transfection of the collecting duct, proximal tubule, distal tubule, interstitial cells, and rarely glomerular cells following injection. To assess renal injury, we performed the renal pelvis injections on uninephrectomised mice and found that their blood urea nitrogen was elevated at two days post-transfer but resolved within two weeks. Although luciferase expression quickly decreased following renal pelvis injection, the use of the piggyBac transposon system improved long-term expression. Immunosuppression with cyclophosphamide stabilised luciferase expression, suggesting immune clearance of the transfected cells occurs in immunocompetent animals. Injection of a transposon expressing erythropoietin raised the haematocrit, indicating that the developed injection technique can elicit a biologic effect in vivo. Hydrodynamic renal pelvis injection enables transposon mediated-kidney specific gene transfer in adult mice.
Department of Veterans Affairs, Nashville, TN 37212 USA.|Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232 USA.|Department of Medicine, Baylor College of Medicine, Houston, TX 77030, USA.
Lauren E
LE
Woodard
Department of Medicine, Baylor College of Medicine, Houston, TX 77030, USA.
Jizhong
J
Cheng
Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232 USA.
Richard C
RC
Welch
Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232 USA.
Felisha M
FM
Williams
Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232 USA.
Wentian
W
Luo
Department of Veterans Affairs, Nashville, TN 37212 USA.|Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232 USA.|Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN 37232 USA.
Leslie S
LS
Gewin
Department of Veterans Affairs, Nashville, TN 37212 USA.|Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232 USA.|Department of Medicine, Baylor College of Medicine, Houston, TX 77030, USA.|Department of Cell and Developmental Biology, Vanderbilt University Medical Center, Nashville, TN 37232 USA.|Department of Pharmacology, Vanderbilt University Medical Center, Nashville, TN 37232 USA.|Department of Veterans Affairs, Houston, TX 77030 USA.
Matthew H
MH
Wilson
D058186:Acute Kidney Injury / Q000209:etiology / Q000473:pathology; D000818:Animals; D004251:DNA Transposable Elements; D004921:Erythropoietin / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005786:Gene Expression Regulation / Q000187:drug effects; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D057446:Hydrodynamics; D007166:Immunosuppressive Agents / Q000494:pharmacology; D007668:Kidney / Q000378:metabolism* / Q000473:pathology; D008297:Male; D051379:Mice; D009928:Organ Specificity; D011401:Promoter Regions, Genetic; D014162:Transfection / Q000379:methods
D004251:DNA Transposable Elements; D007166:Immunosuppressive Agents; D004921:Erythropoietin