CRISPR/Cas9 knockout of USP18 enhances type I IFN responsiveness and restricts HIV-1 infection in macrophages

Taylor, Jared P.; Cash, Melanie N.; Santostefano, Katherine E.; Nakanishi, Mahito; Terada, Naohiro; Wallet, Mark A.

doi:10.1002/jlb.3mia0917-352r

Cited by 47 publications

(54 citation statements)

References 79 publications

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“…These data strongly support a mechanistic model, wherein interferon beta drives the anti-inflammasome and inhibits the inflammasome module through activation of immune suppressors, including Usp18, implicating interferon beta as a potential suppressor of immune activity in the chronic phase of neurodegenerative tauopathies ( Fig. 4E), similar to what has been reported in chronic viral infections 62,65,66 .…”

Section: Identification Of the Inflammasome And Anti-inflammasome Relsupporting

confidence: 85%

“…Several features of the anti-inflammasome module suggest it too may represent aspects of interferonmediated immunosuppression, including its anti-correlation with the inflammasome module, and its inclusion of genes that function as immune checkpoints (Cd274 (PDL1), Il10rb, Lag3) 54,56-58 and inhibitors of immune activity (Usp18 59-62 , Nfkbiz, Nfkbia, Nfkbie, Tgfbr2 63,64 ). Among these genes is Usp18, an established negative feedback suppressor of type 1 interferon anti-viral immune activity [59][60][61][62]65,66 that is also highly connected with the anti-inflammasome module in microglia treated with interferon-beta ( Supplementary Fig. 4G).…”

Section: Identification Of the Inflammasome And Anti-inflammasome Relmentioning

confidence: 99%

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Dementia risk genes engage gene networks poised to tune the immune response towards chronic inflammatory states

Rexach

Swarup

Chang

et al. 2019

Preprint

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An emerging challenge in neurodegenerative dementia is understanding how immune-associated genes and pathways contribute to disease. To achieve a refined view of neuroinflammatory signaling across neurodegeneration, we took an integrative functional genomics approach to consider neurodegeneration from the perspective of microglia and their interactions with other cells. Using large-scale gene expression and perturbation data, regulatory motif analysis, and gene knockout studies, we identify and characterize a microglial-centric network involving distinct gene co-expression modules associated with progressive stages of neurodegeneration. These modules, which are conserved from mouse to human, differentially incorporate specific immune sensors of cellular damage and pathways that are predicted to eventually tune the immune response toward chronic inflammation and immune suppression. Notably, common genetic risk for Alzheimer's disease (AD), Frontotemporal dementia (FTD) and Progressive Supranuclear Palsy (PSP) resides in specific modules that distinguish between the disorders, but also show convergence on pathways related to anti-viral defense mechanisms. These results suggest a model wherein combinatorial microglial-immune signaling integrate specific immune activators and disease genes that lead to the establishment of chronic states of simultaneous inflammation and immunosuppression involving type 1 interferon in these dementias.

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Section: Identification Of the Inflammasome And Anti-inflammasome Relsupporting

confidence: 85%

Section: Identification Of the Inflammasome And Anti-inflammasome Relmentioning

confidence: 99%

Dementia risk genes engage gene networks poised to tune the immune response towards chronic inflammatory states

Rexach

Swarup

Chang

et al. 2019

Preprint

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“…KEYWORDS THP-1, USP18, human immunodeficiency virus 1 (HIV-1), p21, restriction factor C ellular antiretroviral restriction factors are an important component of the innate immune response against HIV-1 (1)(2)(3). Many of these proteins are induced or activated by interferons (IFNs) (4)(5)(6).…”

mentioning

confidence: 99%

“…Experimental knockout of USP18 enhances JAK/STAT (signal transducer and activator of transcription) signaling and increases ISGs with elevated levels of protein ISGylation, thus providing resistance to viral infections (3,42,43,52). Recent work by Honke et al (49) showed in 2012 that high expression of USP18 in murine CD169 ϩ macrophages in the splenic marginal zone was required to enforce a local replication of vesicular stomatitis virus (VSV), a negative-sense, single-stranded enveloped RNA virus belonging to the family Rhabdoviridae.…”

mentioning

confidence: 99%

USP18 (UBP43) Abrogates p21-Mediated Inhibition of HIV-1

Kuffour

Schott

Vasudevan

et al. 2018

J Virol

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The host intrinsic innate immune system drives antiviral defenses and viral restriction, which includes the production of soluble factors, such as type I and III interferon (IFN), and activation of restriction factors, including SAMHD1, a deoxynucleoside triphosphohydrolase. Interferon-stimulated gene 15 (ISG15)-specific ubiquitin-like protease 43 (USP18) abrogates IFN signaling pathways. The cyclin-dependent kinase inhibitor p21 (CIP1/WAF1), which is involved in the differentiation and maturation of monocytes, inhibits human immunodeficiency virus type 1 (HIV-1) in macrophages and dendritic cells. p21 inhibition of HIV-1 replication is thought to occur at the reverse transcription step, likely by suppressing cellular deoxynucleoside triphosphate (dNTP) biosynthesis and increasing the amount of antivirally active form of SAMHD1. SAMHD1 strongly inhibits HIV-1 replication in myeloid and resting CD4 T cells. Here, we studied how USP18 influences HIV-1 replication in human myeloid THP-1 cells. We found that USP18 has the novel ability to inhibit the antiviral function of p21 in differentiated THP-1 cells. USP18 enhanced reverse transcription of HIV-1 by downregulating p21 expression and upregulating intracellular dNTP levels. p21 downregulation by USP18 was associated with the active form of SAMHD1, phosphorylated at T592. USP18 formed a complex with the E3 ubiquitin ligase recognition factor SKP2 (S-phase kinase associated protein 2) and SAMHD1. CRISPR-Cas9 knockout of USP18 increased p21 protein expression and blocked HIV-1 replication. Overall, we propose USP18 as a regulator of p21 antiviral function in differentiated myeloid THP-1 cells. Macrophages and dendritic cells are usually the first point of contact with pathogens, including lentiviruses. Host restriction factors, including SAMHD1, mediate the innate immune response against these viruses. However, HIV-1 has evolved to circumvent the innate immune response and establishes disseminated infection. The cyclin-dependent kinase inhibitor p21, which is involved in differentiation and maturation of monocytes, blocks HIV-1 replication at the reverse transcription step. p21 is thought to suppress key enzymes involved in dNTP biosynthesis and activates SAMHD1 antiviral function. We report here that the human USP18 protein is a novel factor potentially contributing to HIV replication by blocking the antiviral function of p21 in differentiated human myeloid cells. USP18 downregulates p21 protein expression, which correlates with upregulated intracellular dNTP levels and the antiviral inactive form of SAMHD1. Depletion of USP18 stabilizes p21 protein expression, which correlates with dephosphorylated SAMHD1 and a block to HIV-1 replication.

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“…Buffy coats were obtained from Life South Community Blood Center (Gainesville, FL) under approval by the Institutional Review Board at the University of Florida. PBMCs were processed as previously described 54 and CD4 + T cells were enriched with the CD4 + T Cell Isolation Kit (Miltenyi Biotec) according to the manufacturer's instructions. CD4 + T cells were cultured in Advanced RPMI 1640 (Corning) supplemented with 2 mM GlutaMAX (Gibco), 10% heat-inactivated FBS (Sigma-Aldrich, St. Louis, MO), and 100 U/mL penicillin-streptomycin.…”

Section: Primary Cd4 + T Cellsmentioning

confidence: 99%

Harmine enhances the activity of the HIV-1 latency-reversing agents ingenol A and SAHA

Taylor

Armitage

Aldridge

et al. 2020

Preprint

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Infection of HIV-1 remains incurable because long-lived, latently-infected cells persist during prolonged antiretroviral therapy. Attempts to pharmacologically reactivate and purge the latent reservoir with latency reactivating agents (LRAs) such as protein kinase C (PKC) agonists (e.g. ingenol A) or histone deacetylase (HDAC) inhibitors (e.g. SAHA) have shown promising but incomplete efficacy. Using the J-Lat T cell model of HIV latency, we found that the plantderived compound harmine enhanced the efficacy of existing PKC agonist LRAs in reactivating latently-infected cells. Treatment with harmine increased not only the number of reactivated cells but also increased HIV transcription and protein expression on a per-cell basis. Importantly, we observed an additive effect when harmine was used in combination with ingenol A and the HDAC inhibitor SAHA. An investigation into the mechanism revealed that harmine, when used with LRAs, increased the availability of transcription factors needed for viral reactivation such as NFκB, MAPK p38, and ERK1/2. We also found that harmine treatment resulted in reduced expression of HEXIM1, a negative regulator of transcriptional elongation. Despite harmine's reported inhibitory effects on DYRK1A and consequent enhancement of NFAT signaling, the HIV reactivating effects of harmine occurred independent of DYRK1A and NFAT. Harmine increases the efficacy of LRAs by increasing the availability of HIV-1 transcription factors and decreasing expression of HEXIM1. Combination therapies with harmine and LRAs could benefit patients by achieving deeper reactivation of the latent pool of HIV provirus.3 Introduction:

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CRISPR/Cas9 knockout of USP18 enhances type I IFN responsiveness and restricts HIV-1 infection in macrophages

Cited by 47 publications

References 79 publications

Dementia risk genes engage gene networks poised to tune the immune response towards chronic inflammatory states

Dementia risk genes engage gene networks poised to tune the immune response towards chronic inflammatory states

USP18 (UBP43) Abrogates p21-Mediated Inhibition of HIV-1

Harmine enhances the activity of the HIV-1 latency-reversing agents ingenol A and SAHA

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