Comparative Genomics RNAi Screen Identifies Eftud2 as a Novel Regulator of Innate Immunity

Arras, Lesly De; Laws, Rebecca L.; Leach, Sonia M.; Pontis, Kyle; Freedman, Jonathan H.; Schwartz, David A.; Alper, Scott

doi:10.1534/genetics.113.160499

Cited by 52 publications

(47 citation statements)

References 97 publications

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“…EFTUD2 is a GTPase responsible for pre-mRNA splicing and was recently identified as a novel regulator of innate immunity (29). Using a genome-wide siRNA screen, our laboratory recently identified EFTUD2's role in restricting HCV infection.…”

Section: Discussionmentioning

confidence: 99%

EFTUD2 Is a Novel Innate Immune Regulator Restricting Hepatitis C Virus Infection through the RIG-I/MDA5 Pathway

Zhu

Xiao

Hong

et al. 2015

J Virol

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The elongation factor Tu GTP binding domain-containing protein 2 (EFTUD2) was identified as an anti-hepatitis C virus (HCV) host factor in our recent genome-wide small interfering RNA (siRNA) screen. In this study, we sought to further determine EFTUD2's role in HCV infection and investigate the interaction between EFTUD2 and other regulators involved in HCV innate immune (RIG-I, MDA5, TBK1, and IRF3) and JAK-STAT1 pathways. We found that HCV infection decreased the expression of EFTUD2 and the viral RNA sensors RIG-I and MDA5 in HCV-infected Huh7 and Huh7.5.1 cells and in liver tissue from in HCVinfected patients, suggesting that HCV infection downregulated EFTUD2 expression to circumvent the innate immune response. H epatitis C virus (HCV) poses a threat to public health by infecting approximately 170 million people worldwide and causing chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma (1, 2). Fifty to ninety percent of acute infections become chronic due to failure to mount a productive immune response to clear the virus. The innate immune system is the first line of defense responsible for recognizing viral pathogens and is therefore one of the crucial elements in determining the outcome of HCV infection.It is known that HCV RNA is recognized by the combined actions of protein kinase R (PKR), retinoic acid-inducible gene 1 (RIG-I), and Toll-like receptor 3 (TLR3) after viral entry into host cells (3-5). Pathogen recognition triggers downstream signaling to activate transcription factors, such as interferon (IFN) regulatory factors (IRFs), to induce type I interferon secretion and stimulation of the JAK-STAT signaling pathway, leading to the expression of IFN-stimulated genes (ISGs), which presumptively mediate control of viral replica-

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Section: Discussionmentioning

confidence: 99%

EFTUD2 Is a Novel Innate Immune Regulator Restricting Hepatitis C Virus Infection through the RIG-I/MDA5 Pathway

Zhu

Xiao

Hong

et al. 2015

J Virol

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“…Transient transfections were performed as previously described (De Arras et al 2014b). Briefly, 100,000 cells plated in 24-well format were transfected with 3.75 ml Fugene HD (Roche), 300 ng NF-kB-AP1-luc (a derivative of the IL-8 promoter) (Mukaida et al 1989), 100 ng SV40-rluc (Promega, Madison, WI), and 600 ng of overexpression plasmid.…”

Section: Whole Genome Association Mapping Analysismentioning

confidence: 99%

Novel Innate Immune Genes Regulating the Macrophage Response to Gram Positive Bacteria

et al. 2016

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Host variation in Toll-like receptors and other innate immune signaling molecules alters infection susceptibility. However, only a portion of the variability observed in the innate immune response is accounted for by known genes in these pathways. Thus, the identification of additional genes that regulate the response to Gram positive bacteria is warranted. Bone marrow-derived macrophages (BMMs) from 43 inbred mouse strains were stimulated with lipotechoic acid (LTA), a major component of the Gram positive bacterial cell wall. Concentrations of the proinflammatory cytokines IL-6, IL-12, and TNF-a were measured. In silico whole genome association (WGA) mapping was performed using cytokine responses followed by network analysis to prioritize candidate genes. To determine which candidate genes could be responsible for regulating the LTA response, candidate genes were inhibited using RNA interference (RNAi) and were overexpressed in RAW264.7 macrophages. BMMs from Bdkrb1-deficient mice were used to assess the effect of Bdkrb1 gene deletion on the response to LTA, heat-killed Streptococcus pneumoniae, and heat-killed Staphylococcus aureus. WGA mapping identified 117 loci: IL-6 analysis yielded 20 loci (average locus size = 0.133 Mb; 18 genes), IL-12 analysis produced 5 loci (0.201 Mb average; 7 genes), and TNF-a analysis yielded 92 loci (0.464 Mb average; 186 genes of which 46 were prioritized by network analysis). The follow-up small interfering RNA screen of 71 target genes identified four genes (Bdkrb1, Blnk, Fbxo17, and Nkx6-1) whose inhibition resulted in significantly reduced cytokine production following LTA stimulation. Overexpression of these four genes resulted in significantly increased cytokine production in response to LTA. Bdkrb1-deficient macrophages were less responsive to LTA and heat-killed S. aureus, validating the genetic and RNAi approach to identify novel regulators of the response to LTA. We have identified four innate immune response genes that may contribute to Gram positive bacterial susceptibility.KEYWORDS innate immunity; Gram positive bacteria; lipotechoic acid; whole genome association mapping; inbred strains of mice; RNA interference G RAM positive bacterial infections are a major public health concern, with pneumonia for example, being the most prevalent lower respiratory infection and third leading cause of death around the world (Liu et al. 2015). Around 10% of healthy adults and 20-40% of healthy children have airways colonized by the most common pneumonia pathogen, Streptococcus pneumoniae (van der Poll and Opal 2009) but only a small portion of these individuals become actively infected (Brouwer et al. 2009). Similarly, the rate of Staphylococcus aureus infection is high in hospitals and community settings. However, animal and human studies have shown variable host response and course of disease (Shukla et al. 2015), suggesting genetic factors in the innate immune response to the pathogen are likely at play.The innate immune response begins with binding of the pathogen-as...

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“…[13][14][15] Among these factors, the mRNA splicing regulator Eftud2, whose mutation causes a multiple malformation syndrome termed to mandibulofacial dysostosis with microcephaly, 16 was recently shown to control the alternate splicing of the MyD88 adaptor to modulate the extent of the innate immune response in murine macrophages. 17 However, the precise role of Eftud2 in chronic intestinal inflammation and the associated tumorigenesis is yet unknown.…”

Section: Introductionmentioning

confidence: 99%

Spliceosome protein Eftud2 promotes colitis-associated tumorigenesis by modulating inflammatory response of macrophage

Lv¹,

Wang

Luo³

et al. 2019

Mucosal Immunology

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Alternative splicing (AS) of mRNA is known to be involved in regulation of immune cell differentiation and activation. Elongation factor Tu GTP binding domain containing 2 (Eftud2) is an AS factor to potentially modulate innate immune response in macrophages. In this study, we investigate its involvement in the pathogenesis of colitis-associated cancer (CAC). Using an established mouse model of CAC, we show that Eftud2 is constantly overexpressed in the colonic tissues as well as infiltrating macrophages. Myeloid-specific knockout of Eftud2 remarkably suppresses chronic intestinal inflammation and tumorigenesis, which is associated with decreased production of inflammatory cytokines and tumorigenic factors. Repression of colonic inflammation and colorectal tumor development in Eftud2-deficient mice is due to the impaired activation of NF-κB signaling in LPS-challenged macrophages. Furthermore, the alteration of Eftud2-mediated AS involving the components of TLR4-NF-κB cascades underlies the impairment of NF-κB activation. Overall, these findings provide new insights into the tight link between inflammation and cancer and modulation of AS in innate immune signals may be a potentially therapeutic avenue for CAC treatment.

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Comparative Genomics RNAi Screen Identifies Eftud2 as a Novel Regulator of Innate Immunity

Cited by 52 publications

References 97 publications

EFTUD2 Is a Novel Innate Immune Regulator Restricting Hepatitis C Virus Infection through the RIG-I/MDA5 Pathway

EFTUD2 Is a Novel Innate Immune Regulator Restricting Hepatitis C Virus Infection through the RIG-I/MDA5 Pathway

Novel Innate Immune Genes Regulating the Macrophage Response to Gram Positive Bacteria

Spliceosome protein Eftud2 promotes colitis-associated tumorigenesis by modulating inflammatory response of macrophage

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