Genome‐wide CRISPR‐Cas9 viability screen reveals genes involved in TNF‐α‐induced apoptosis of human umbilical vein endothelial cells
Journal of Cellular Physiology
Published online on October 14, 2018
Abstract
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- "\nAbstract\nTumor necrosis factor α (TNF‐α), a pivotal cytokine in sepsis, protects
the host against pathogens by promoting an inflammatory response while simultaneously
inducing apoptosis of the vascular endothelium. Unfortunately, inhibitors targeting
certain components of the TNF‐α signaling pathway to reduce cellular apoptosis have
failed to translate into clinical applications, partly due to the adverse effects
of excessive immunosuppression. In an attempt to discover potential targets in the
TNF‐α signaling pathway to modulate moderate inflammation and apoptosis during the
development of sepsis, we performed a pooled genome‐wide CRISPR/Cas9 knockout screen
in human umbilical vein endothelial cells (HUVECs). Tumor necrosis factor receptor
superfamily member 1A (TNFRSF1A), B‐cell lymphoma 2 (BCL2), Bcl2‐associated death
promoter (BAD), and NLR family member X1 (NLRX1) deficiencies were identified as
the effective genetic suppressors of TNF‐α cytotoxicity on a list of candidate regulators.
CRISPR‐mediated \nNLRX1 knockout conferred cellular resistance to challenge with
TNF‐α, and NLRX1 could be induced to colocalize with mitochondria following TNF‐α
stimulation. Thus, our work demonstrates the advantage of genome‐scale screening
with Cas9 and validates NLRX1 as a potential modulator of TNF‐α‐induced vascular
endothelial apoptosis during sepsis."
- Journal of Cellular Physiology, EarlyView.