title: Targeted delivery of AAV-transduced mesenchymal stromal cells to hepatic tissue for ex vivo gene therapy.
abstract: Adeno-associated virus (AAV)-mediated gene therapy holds great promise if challenges related to vector neutralization by pre-existing antibodies are circumvented. The use of autologous or allogeneic cells to shield the vector might offer the possibility of successful gene transfer in such a situation. In the present study, we evaluated the feasibility of AAV-transduced mesenchymal stromal cells (MSCs) as a vehicle for hepatic gene transfer in a murine liver injury model. In our initial studies to determine the most suitable vector, we observed that AAV1 (91%) and AAV6 (72%) serotypes are highly efficient in transducing MSCs. Subsequently, we generated a transient liver injury model to analyse the efficacy of MSCs homing to the liver, as well as their hepatic gene transfer efficiency; our data show that administration of acetaminophen (500 mg/kg) served as a cue for the homing of MSCs to the liver. Furthermore, sex-mismatched transplantation of AAV1-infected MSCs demonstrated a 3.5-fold (day 7) and 2.2-fold (day 28) higher hepatic gene transfer efficiency. To further corroborate this, we estimated the donor cell Y chromosome copies in the liver of recipient female mice. Our data revealed a 12.7-fold increase in average genome copies of male MSCs in the livers of recipient mice with injury compared to control, 60 days after transplantation. However, in vivo administration of AAV-transduced MSCs in the presence of neutralization antibodies (intravenous immunoglobulin, IVIG) was not beneficial. This is possibly due to the clearance of transplanted MSCs by circulating IVIG and underscores the need to develop suitable in vivo models to study such a mode of gene transfer. Copyright © 2015 John Wiley & Sons, Ltd.
authors.affiliation: Department of Haematology, Christian Medical College, Vellore, Tamil Nadu, India.
authors.forename: Nishanth
authors.initials: N
authors.lastname: Gabriel
authors.affiliation: Centre for Stem Cell Research, Christian Medical College, Vellore, Tamil Nadu, India.
authors.forename: Rekha
authors.initials: R
authors.lastname: Samuel
authors.affiliation: Department of Haematology, Christian Medical College, Vellore, Tamil Nadu, India.|Department of Biological Sciences and Bioengineering, Indian Institute of Technology, Kanpur, Uttar Pradesh, India.
authors.forename: Giridhara R
authors.initials: GR
authors.lastname: Jayandharan
mesh_terms: D000082:Acetaminophen / Q000009:adverse effects / Q000494:pharmacology; D064591:Allografts; D000818:Animals; D056486:Chemical and Drug Induced Liver Injury / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D000229:Dependovirus; D015316:Genetic Therapy; D008099:Liver / Q000378:metabolism* / Q000473:pathology; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D014161:Transduction, Genetic
chemical_list: D000082:Acetaminophen
keywords: adeno-associated virus; gene therapy; hepatocyte; liver injury model; mesenchymal stromal cell; viral vectors