Adenoviral vector encoding soluble Flt-1 engineered human endometrial mesenchymal stem cells effectively regress endometriotic lesions in NOD/SCID mice

This study was undertaken to study the efficiency of Adsflt-1 engineered human eutopic mesenchymal stem cells (MSCs) secreting anti-angiogenic sFlt-1 as a targeted cell-based therapy for endometriosis (EM). Eutopic MSCs were transduced with Adsflt-1/AdV0 viral vectors and were evaluated for expression and secretion of sFlt-1. EM was created in NOD/SCID mice using subcutaneous implantation techniques. Four doses of 106 MSC-Adsflt-1/MSC-AdV0 were administered to the model and therapeutic anti-angiogenic ability was analyzed by lesion size measurement, microvessel density, immunohistochemistry and real-time reverse transcriptase-PCR analysis. Approximately 86% of transduced MSCs expressed and secreted sFlt-1. MSC-Adsflt-1-treated animals exhibited significant reduction (52.8±1.8%) in size of endometriotic lesions. We observed a 2.3-fold decrease in the number and a 10-fold decrease in the size of endometrial glands in MSC-Adsflt-1-treated animals. A two-fold decrease in stromal cell densities was also observed in MSC-Adsflt-1-treated animals compared with the MSC-AdV0 group. Specific positive immunostaining for MSC marker, CD146 and sFlt-1 in the lesion sites of the MSC-Adsflt-1 group suggests possible homing of transduced MSCs, their survival and secretion of sFlt-1 at the target sites. A marked reduction in size of microvessels and microvessel density within endometriotic lesions and surrounding host subcutaneous layers was observed in MSC-Adsflt-1 group along with significantly downregulated expression of transcripts for vascular endothelial growth factor, fetal liver kinase 1 and matrix metalloproteinases (2 and 9). Our findings indicate the efficacy of a novel eutopic MSC-Adsflt-1 therapy in EM study models. Evaluating long-term effects of genetically modified MSCs in vivo is essential in translating MSC-Adsflt-1 therapy to the clinics. This is a preview of subscription content, access via your institution Access options Subscribe to this journal Receive 6 print issues and online access 251,40 € per year only 41,90 € per issue Buy this article - Purchase on SpringerLink - Instant access to the full article PDF. 39,95 € Prices may be subject to local taxes which are calculated during checkout Similar content being viewed by others

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We are grateful to Dr Ronald Crystal for being generous in providing the Adsflt-1 vector. We also thank Dr SD Mahale, Director, NIRRH, Mumbai for providing necessary facilities for research. Ms Sushama, Ms Gayatri and Dr S Mukherjee are acknowledged for their help in flow cytometry and Ms Reshma, Ms Shobha and Dr N. Balasinor for help in confocal microscopy. We thank Mr Vaibhav, graphics expert, for his help in improving the resolution of figures. This work was supported by the Post-doctoral fellowship program of the Indian Council of Medical Research, New Delhi, India. Author information Authors and Affiliations Corresponding author Ethics declarations Competing interests The authors declare no conflict of interest. Additional information Supplementary Information accompanies this paper on Gene Therapy website Rights and permissions About this article Cite this article Koippallil Gopalakrishnan, A., Pandit, H., Metkari, S. et al. Adenoviral vector encoding soluble Flt-1 engineered human endometrial mesenchymal stem cells effectively regress endometriotic lesions in NOD/SCID mice. Gene Ther 23, 580–591 (2016). https://doi.org/10.1038/gt.2016.30 Received: Revised: Accepted: Published: Issue date: DOI: https://doi.org/10.1038/gt.2016.30