Generation of Human Inflammation-Resistant Endothelial Progenitor Cells by A20 Gene Transfer

Abstract

Inflammatory activation of the vascular endothelium is a major contributory factor to ischemic cardiovascular disease. Endothelial progenitor cells (EPCs) are being investigated for the treatment of ischemic disease or to coat vein grafts for bypass surgery. As an inflammatory environment might reduce their therapeutic efficacy, we sought to generate EPCs that are less sensitive to inflammatory activation. EPCs were obtained from human umbilical cord blood and transduced with a lentiviral vector for stable expression of A20, an anti-inflammatory protein. Nontransduced and green-fluorescent-protein-transduced cells were used as controls. Expression of A20 by EPCs did not modify cell morphology or expression of a panel of 20 proteins known to contribute to angiogenesis. Also, A20 had no effect on the capacity of EPCs to form tube-like structures in Matrigel™. A20 expression reduced EPC activation by tumor necrosis factor-α and interleukin-1β as determined from changes in vascular cell adhesion molecule 1 and E-selectin expression and decreased monocyte transmigration through a monolayer of EPCs. In conclusion, EPCs can be genetically modified to overexpress A20 in a stable fashion. These cells become less sensitive to inflammatory stimuli. This may be of interest in cell-based therapeutic approaches for clinical settings where inflammation is an important pathogenic factor.

Induction of Vasculogenic Mimicry Overrides VEGF-A Silencing and Enriches Stem-like Cancer Cells in Melanoma

“…Real-time quantitative PCR (qRT-PCR) was performed subsequently on a StepOnePlus™ Real-Time PCR System (Applied Biosystems, Foster City, CA) using human-specific primers. The primer sequences were as follows: CD133 forward 5’-TTCTTGACCGACTGAGAC-3’ and reverse 5’-CCAAGCACAGAGGGTCAT-3’; CD144 forward 5’-ATATGTCAGTGATGACTA-3’ and reverse 5’-CTTACCAGGGCGTTCAGG-3’; CD271 forward 5’-ACTCACTGCACAGACTCT-3’ and reverse 5’-GAAGCTTCTCAACGGCTC-3’; MMP-2 forward 5’-TTTCCATTCCGCTTCCAGGGCAC-3’ and reverse 5’-TCGCACACCACATCTTTCCGTCACT-3’ (24); Tie-1 forward 5’-CACGACCATGACGGCGAAT-3’ and reverse 5’-CGGCAGCCTGATATGCCTG-3’ (25); and GAPDH forward 5'-CGACCACTTTGTCAAGCTCA-3' and reverse 5'-AGGGGAGATTCAGTGTGGTG-3’. All samples were run in triplicate and normalized to the housekeeping gene, GAPDH.…”

Dysfunctional Endothelial Progenitor Cells in Chronic Kidney Disease

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Involvement of endothelial progenitor cells in the formation of plexiform lesions in broiler chickens: possible role of local immune/inflammatory response

“…Our results also suggest that the local immune cell infiltrate may contribute to the development of PLs by inducing EPC death and limiting their response to angiogenic stimuli. Given the therapeutic potential of EPCs in repairing vascular damage (Sukmawati and Tanaka, 2015), transplantation of gene-modified inflammationresistant EPCs (Liu et al, 2010) may be beneficial for intervention of the formation of PLs. Further studies are warranted to test this hypothesis.…”