A Genome-Wide Knockout Screen in Human Macrophages Identified Host Factors Modulating <italic toggle="yes">Salmonella</italic> Infection

ABSTRACT A genome-scale CRISPR knockout library screen of THP-1 human macrophages was performed to identify loss-of-function mutations conferring resistance to Salmonella uptake. The screen identified 183 candidate genes, from which 14 representative genes involved in actin dynamics (ACTR3, ARPC4, C...

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Autores principales:	Amy T. Y. Yeung, Yoon Ha Choi, Amy H. Y. Lee, Christine Hale, Hannes Ponstingl, Derek Pickard, David Goulding, Mark Thomas, Erin Gill, Jong Kyoung Kim, Allan Bradley, Robert E. W. Hancock, Gordon Dougan
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	CRISPR Salmonella bacteria genome-wide screen macrophages Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/88e1f65dbd364ffc8b38e80865afebb5
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spelling	oai:doaj.org-article:88e1f65dbd364ffc8b38e80865afebb52021-11-15T15:59:42ZA Genome-Wide Knockout Screen in Human Macrophages Identified Host Factors Modulating <italic toggle="yes">Salmonella</italic> Infection10.1128/mBio.02169-192150-7511https://doaj.org/article/88e1f65dbd364ffc8b38e80865afebb52019-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02169-19https://doaj.org/toc/2150-7511ABSTRACT A genome-scale CRISPR knockout library screen of THP-1 human macrophages was performed to identify loss-of-function mutations conferring resistance to Salmonella uptake. The screen identified 183 candidate genes, from which 14 representative genes involved in actin dynamics (ACTR3, ARPC4, CAPZB, TOR3A, CYFIP2, CTTN, and NHLRC2), glycosaminoglycan metabolism (B3GNT1), receptor signaling (PDGFB and CD27), lipid raft formation (CLTCL1), calcium transport (ATP2A2 and ITPR3), and cholesterol metabolism (HMGCR) were analyzed further. For some of these pathways, known chemical inhibitors could replicate the Salmonella resistance phenotype, indicating their potential as targets for host-directed therapy. The screen indicated a role for the relatively uncharacterized gene NHLRC2 in both Salmonella invasion and macrophage differentiation. Upon differentiation, NHLRC2 mutant macrophages were hyperinflammatory and did not exhibit characteristics typical of macrophages, including atypical morphology and inability to interact and phagocytose bacteria/particles. Immunoprecipitation confirmed an interaction of NHLRC2 with FRYL, EIF2AK2, and KLHL13. IMPORTANCE Salmonella exploits macrophages to gain access to the lymphatic system and bloodstream to lead to local and potentially systemic infections. With an increasing number of antibiotic-resistant isolates identified in humans, Salmonella infections have become major threats to public health. Therefore, there is an urgent need to identify alternative approaches to anti-infective therapy, including host-directed therapies. In this study, we used a simple genome-wide screen to identify 183 candidate host factors in macrophages that can confer resistance to Salmonella infection. These factors may be potential therapeutic targets against Salmonella infections.Amy T. Y. YeungYoon Ha ChoiAmy H. Y. LeeChristine HaleHannes PonstinglDerek PickardDavid GouldingMark ThomasErin GillJong Kyoung KimAllan BradleyRobert E. W. HancockGordon DouganAmerican Society for MicrobiologyarticleCRISPRSalmonellabacteriagenome-wide screenmacrophagesMicrobiologyQR1-502ENmBio, Vol 10, Iss 5 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	CRISPR Salmonella bacteria genome-wide screen macrophages Microbiology QR1-502
spellingShingle	CRISPR Salmonella bacteria genome-wide screen macrophages Microbiology QR1-502 Amy T. Y. Yeung Yoon Ha Choi Amy H. Y. Lee Christine Hale Hannes Ponstingl Derek Pickard David Goulding Mark Thomas Erin Gill Jong Kyoung Kim Allan Bradley Robert E. W. Hancock Gordon Dougan A Genome-Wide Knockout Screen in Human Macrophages Identified Host Factors Modulating <italic toggle="yes">Salmonella</italic> Infection
description	ABSTRACT A genome-scale CRISPR knockout library screen of THP-1 human macrophages was performed to identify loss-of-function mutations conferring resistance to Salmonella uptake. The screen identified 183 candidate genes, from which 14 representative genes involved in actin dynamics (ACTR3, ARPC4, CAPZB, TOR3A, CYFIP2, CTTN, and NHLRC2), glycosaminoglycan metabolism (B3GNT1), receptor signaling (PDGFB and CD27), lipid raft formation (CLTCL1), calcium transport (ATP2A2 and ITPR3), and cholesterol metabolism (HMGCR) were analyzed further. For some of these pathways, known chemical inhibitors could replicate the Salmonella resistance phenotype, indicating their potential as targets for host-directed therapy. The screen indicated a role for the relatively uncharacterized gene NHLRC2 in both Salmonella invasion and macrophage differentiation. Upon differentiation, NHLRC2 mutant macrophages were hyperinflammatory and did not exhibit characteristics typical of macrophages, including atypical morphology and inability to interact and phagocytose bacteria/particles. Immunoprecipitation confirmed an interaction of NHLRC2 with FRYL, EIF2AK2, and KLHL13. IMPORTANCE Salmonella exploits macrophages to gain access to the lymphatic system and bloodstream to lead to local and potentially systemic infections. With an increasing number of antibiotic-resistant isolates identified in humans, Salmonella infections have become major threats to public health. Therefore, there is an urgent need to identify alternative approaches to anti-infective therapy, including host-directed therapies. In this study, we used a simple genome-wide screen to identify 183 candidate host factors in macrophages that can confer resistance to Salmonella infection. These factors may be potential therapeutic targets against Salmonella infections.
format	article
author	Amy T. Y. Yeung Yoon Ha Choi Amy H. Y. Lee Christine Hale Hannes Ponstingl Derek Pickard David Goulding Mark Thomas Erin Gill Jong Kyoung Kim Allan Bradley Robert E. W. Hancock Gordon Dougan
author_facet	Amy T. Y. Yeung Yoon Ha Choi Amy H. Y. Lee Christine Hale Hannes Ponstingl Derek Pickard David Goulding Mark Thomas Erin Gill Jong Kyoung Kim Allan Bradley Robert E. W. Hancock Gordon Dougan
author_sort	Amy T. Y. Yeung
title	A Genome-Wide Knockout Screen in Human Macrophages Identified Host Factors Modulating <italic toggle="yes">Salmonella</italic> Infection
title_short	A Genome-Wide Knockout Screen in Human Macrophages Identified Host Factors Modulating <italic toggle="yes">Salmonella</italic> Infection
title_full	A Genome-Wide Knockout Screen in Human Macrophages Identified Host Factors Modulating <italic toggle="yes">Salmonella</italic> Infection
title_fullStr	A Genome-Wide Knockout Screen in Human Macrophages Identified Host Factors Modulating <italic toggle="yes">Salmonella</italic> Infection
title_full_unstemmed	A Genome-Wide Knockout Screen in Human Macrophages Identified Host Factors Modulating <italic toggle="yes">Salmonella</italic> Infection
title_sort	genome-wide knockout screen in human macrophages identified host factors modulating <italic toggle="yes">salmonella</italic> infection
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/88e1f65dbd364ffc8b38e80865afebb5
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A Genome-Wide Knockout Screen in Human Macrophages Identified Host Factors Modulating <italic toggle="yes">Salmonella</italic> Infection

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