A Systematic Approach to the Establishment and Characterization of Endothelial Progenitor Cells for Gene Therapy

Abstract

It has been recently demonstrated that endothelial progenitor cells (EPCs) have increasing potential for gene therapy or regenerative cell therapy for cardiovascular diseases and cancer. However, current therapies involving EPCs are inefficient because of the very low level of EPCs in the available sources, for example, in blood. One solution is to derive in vitro an expanded population of EPCs from circulation. In addition, EPCs like other progenitor cells have an intrinsic predisposition of differentiating into mature cell types, for example, mature endothelial cells; therefore, establishing a sufficient amount of EPCs alongside maintaining the EPC characteristic phenotype during genetic modification and long-term culture presents a significant challenge to the field of gene and cell therapies. In this study, we have systematically investigated EPCs from different sources and used multiple parameters, including cell surface markers and a tubule formation assay to identify factors that influence the establishment, characteristics, and vector transduction capability of EPCs. Our results show the considerable promise, as well as certain limitations in the establishment and manipulation of genetically modified EPCs for gene therapy. While obtaining high transduction efficiency and robust in vitro tubule formation of EPCs using lentiviral vectors, we also observed that lentiviral vector transduction significantly altered EPC phenotype as demonstrated by an increased percentage of CD34 + progenitor cells and increased expression of adhesion molecule CD144 (VE-cadherin). Taking account of the increased expression of CD144 reported in cancer patients, the altered expression of EPC-related markers, for example, VE-cadherin and the enrichment of CD34 + cells, after vector transduction indicates the importance of extensive characterization and vigorous safety control of genetically modified EPCs before they are accepted for clinical use.

Pharmaceutical Activation of Nrf2 Accelerates Diabetic Wound Healing by Exosomes from Bone Marrow Mesenchymal Stem Cells

“…Late EPCs have a high proliferative capacity and can form vascular tubes in Matrigel in vitro . These cells are typically identified by the coexpression of CD31 and CD34 and negative expression of CD45 ( 25 ). As indicated by the flow cytometric analysis, a vast majority of passage-3 cells showed positive expression of CD31 (98.5%) and CD34 (96.8%), but negative expression of CD45 (1.27%) ( Fig.…”

Capsid Engineering Overcomes Barriers Toward Adeno-Associated Virus Vector-Mediated Transduction of Endothelial Cells

“…50 In addition, EPC home to damaged vasculature, making them promising targets for genetic modification to develop novel therapeutic strategies in the context of regenerative medicine, gene therapy, organ transplantation, and cardiovascular disease 54 as well as antitumor therapy. 55 Given the recent advancement in iPSC technology, in particular, with regard to ''off the shelf cell products,'' iPSC-derived EPC will likely replace conventional EPC in the clinical setting. With its impressive efficacy in transducing iPSC-derived EPC, AAV-V EC may represent a highly potent delivery system also in this setting.…”

Research Status of Tissue Engineering of Endovascular Stent and Spring Ring

“…DA Dichet [6] by using retrovirus mediated gene delivery method, the human tissue plasminogen activator gene was inserted into sheep endothelial cells, endothelial cells were cultured in vitro, and the number of cells attached to the scaffold and secreted high levels of tissue type plasminogen activator. He thinks stent implantation coating of genetically modified endothelial cells, the endothelial cells directly into the artery wall, and the expression of anticoagulant, antithrombotic, anti proliferative molecules to prevent complications after stent implantation.Werling NJ [7] after the recombinant retrovirus recombinant of endothelial progenitor cells and the gene of the protection of the gene of the vascular protection of rabbit endothelial progenitor cells and encoding, the common carotid artery after balloon angioplasty. After 2 weeks, the expression product of the gene was detected by the animal blood vessel, the endothelial cell nitric oxide synthase, heme and green fluorescent protein, and the local free of thrombosis, the new intima was small and the proliferation was lower than that in the control group.…”