Development of small molecule and CRISPR based approaches in endothelial cell growth

Endothelial cells are an essential component of the vascular system and became a great candidate for cell-based therapies in the manner of cancer, vascular diseases, and dysfunction as atherosclerosis, hypertension, diabetes, peripheral and cardiovascular diseases. There are novel therapeutic approaches to solve these conditions which involve the development of tissue-engineered veins with ex vivo expanded endothelial cells, development of antiangiogenic therapeutic molecules, and gene therapies with novel approaches. The development of tissue-engineered veins is limited due to the high cell number requirement to accelerate the expansion of vascular forming cells. We have screened small molecules and found that CASIN and AMD3100 increase endothelial expansion, induce tube formation, increase cell migration, and upregulate key endothelial marker genes. In the manner of the development of antiangiogenic therapeutic molecules, we hypothesized that SC1 from screened molecules could be used to inhibit endothelial growth and prevent angiogenesis of endothelial cells due to its antiangiogenic effect on tumor growth. We have demonstrated that SC1 decrease endothelial proliferation, prevent tube formation, increase apoptosis, and upregulate key apoptosis-related genes. In the manner of gene therapy, we hypothesized that CRISPR/Cas9 system could be used to provide a platform for the treatment of various diseases related to endothelial as human immunodeficiency virus (HIV) infection. We have indicated a basic and successful transfection system and on target mutagenesis of CCR5 based on the endonuclease assay. These findings suggest that endothelial related diseases could be solved from the perspective of different approaches. Growth inhibition study of endothelial contributes to the understanding of SC1 treatment in clinical studies in tumor microenvironments. AMD3100 and CASIN treatment can be used to handle high cell number requirements of endothelial cells and to improve new blood vessels ex vivo. Endothelial diseases originated genetically or by infection could be solved with the platform as defined in CRISPR/Cas9 based gene therapy for endothelial in the manner of CCR5 targeting.