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High-Throughput CRISPR Screens To Dissect Macrophage- Shigella Interactions

Authors
  • Lai, Yong
  • Cui, Liang
  • Babunovic, Gregory H.
  • Fortune, Sarah M.
  • Doench, John G.
  • Lu, Timothy K.
Type
Published Article
Journal
mBio
Publisher
American Society for Microbiology
Publication Date
Dec 21, 2021
Volume
12
Issue
6
Identifiers
DOI: 10.1128/mBio.02158-21
PMID: 34933448
PMCID: PMC8689513
Source
PubMed Central
Keywords
License
Unknown

Abstract

Shigellosis causes most diarrheal deaths worldwide, particularly affecting children. Shigella invades and replicates in the epithelium of the large intestine, eliciting inflammation and tissue destruction. To understand how Shigella rewires macrophages prior to epithelium invasion, we performed genome-wide and focused secondary CRISPR knockout and CRISPR interference (CRISPRi) screens in Shigella flexneri -infected human monocytic THP-1 cells. Knockdown of the Toll-like receptor 1/2 signaling pathway significantly reduced proinflammatory cytokine and chemokine production, enhanced host cell survival, and controlled intracellular pathogen growth. Knockdown of the enzymatic component of the mitochondrial pyruvate dehydrogenase complex enhanced THP-1 cell survival. Small-molecule inhibitors blocking key components of these pathways had similar effects; these were validated with human monocyte-derived macrophages, which closely mimic the in vivo physiological state of macrophages postinfection. High-throughput CRISPR screens can elucidate how S. flexneri triggers inflammation and redirects host pyruvate catabolism for energy acquisition before killing macrophages, pointing to new shigellosis therapies.

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