Small molecule inhibition of human cGAS reduces total cGAMP output and cytokine expression in cells

Wiser, Caroline; Kim, Byungil; Vincent, Jessica; Ascano, Manuel

doi:10.1101/2020.03.30.016535

Cited by 6 publications

(6 citation statements)

References 58 publications

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“…al., we observe HIV-1 infection induces cGAS signaling in a NONO-dependent manner in MDDCs (Figures 4A and S4A). However, we find that NONO depletion in THP-1 cells, which maintain an intact innate response to HIV (Collins et al, 2015; Gao et al, 2013; Sumner et al, 2020; Sun et al, 2013; Wiser et al, 2020; Yoh et al, 2015), has no significant effect on innate immune response to HIV-1(with Vpx), as well as HIV-2, infection (Figures 4B). In attempt to determine whether NONO participates in the initial recognition of incoming virus particles in MDDCs, we evaluated the frequency of NONO association with incoming HIV-1 virions.…”

Section: Resultsmentioning

confidence: 88%

Recognition of HIV-1 Capsid Licenses Innate Immune Response to Viral Infection

Yoh

Mamede

Lau

et al. 2022

Preprint

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Cyclic GMP-AMP synthase (cGAS) is a primary sensor of aberrant DNA that governs an innate immune signaling cascade, leading to the induction of the type-I interferon response. We have previously identified polyglutamine binding protein 1, PQBP1, as an adaptor molecule required for cGAS-mediated innate immune response of lentiviruses, including the human immunodeficiency virus 1 (HIV-1), but dispensable for the recognition of DNA viruses. HIV-1-encoded DNA is synthesized as a single copy from its RNA genome, and is subsequently integrated into the host chromatin. HIV-1 then produces progeny through amplification and packaging of its RNA genome, thus, in contrast to DNA viruses, HIV-1 DNA is both transient and of low abundance. However, the molecular basis for the detection and verification of this low abundance HIV-1 DNA pathogen-associated molecular pattern (PAMP) is not understood. Here, we elucidate a two-factor authentication strategy that is employed by the innate immune surveillance machinery to selectively respond to the low concentration of PAMP, while discerning these species from extranuclear DNA molecules. We find that, upon HIV-1 infection, PQBP1 decorates intact viral capsid, which serves as a primary verification step for the viral nucleic acid cargo. As the reverse transcription and capsid disassembly initiate, cGAS protein is then recruited to the capsid in a PQBP1-dependent manner, enabling cGAS molecules to be co-positioned at the site of PAMP generation. Thus, these data indicate that PQBP1 recognition of the HIV-1 capsid sanctions a robust cGAS-dependent response to a limited abundance and short-lived DNA PAMP. Critically, this illuminates a molecular strategy wherein the modular recruitment of co-factors to germline encoded pattern recognition receptors (PRRs) serves to enhance repertoire of pathogens that can be sensed by the innate immune surveillance machinery.

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Section: Resultsmentioning

confidence: 88%

Recognition of HIV-1 Capsid Licenses Innate Immune Response to Viral Infection

Yoh

Mamede

Lau

et al. 2022

Preprint

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“…The role of cGAS in the immunostimulatory activity of the compounds was investigated in the RAW-Dual reporter cells by pretreating the cells with a dose response of the established small molecule inhibitor of cGAS, RU.521 (33,61,62) (Figure 2G). Four hours after incubation with RU.521, the cells were treated with the EC75 concentration of 95-BP/D-PDB (i.e.…”

Section: Resultsmentioning

confidence: 99%

Pharmacological Activation of cGAS for Cancer Immunotherapy

et al. 2021

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When compartmentally mislocalized within cells, nucleic acids can be exceptionally immunostimulatory and can even trigger the immune-mediated elimination of cancer. Specifically, the accumulation of double-stranded DNA in the cytosol can efficiently promote antitumor immunity by activating the cGAMP synthase (cGAS) / stimulator of interferon genes (STING) cellular signaling pathway. Targeting this cytosolic DNA sensing pathway with interferon stimulatory DNA (ISD) is therefore an attractive immunotherapeutic strategy for the treatment of cancer. However, the therapeutic activity of ISD is limited by several drug delivery barriers, including susceptibility to deoxyribonuclease degradation, poor cellular uptake, and inefficient cytosolic delivery. Here, we describe the development of a nucleic acid immunotherapeutic, NanoISD, which overcomes critical delivery barriers that limit the activity of ISD and thereby promotes antitumor immunity through the pharmacological activation of cGAS at the forefront of the STING pathway. NanoISD is a nanoparticle formulation that has been engineered to confer deoxyribonuclease resistance, enhance cellular uptake, and promote endosomal escape of ISD into the cytosol, resulting in potent activation of the STING pathway via cGAS. NanoISD mediates the local production of proinflammatory cytokines via STING signaling. Accordingly, the intratumoral administration of NanoISD induces the infiltration of natural killer cells and T lymphocytes into murine tumors. The therapeutic efficacy of NanoISD is demonstrated in preclinical tumor models by attenuated tumor growth, prolonged survival, and an improved response to immune checkpoint blockade therapy.

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“…Later studies showed that docking of Ru.521 into the active site of human cGAS (Zhou et al, 2018) is suboptimal and the compound therefore has reduced activity against recombinant human cGAS (Lama et al, 2019). Recently, a study found comparable activity of Ru.521 against both mouse and human cGAS in cellular assays (Wiser et al, 2020). X-ray crystal structures of cGAS in complex with Ru.521 have been solved (mouse cGAS: PDB 5XZG; human cGAS: PDB 6O47).…”

Section: Discussionmentioning

confidence: 99%

Regulation and inhibition of the DNA sensor cGAS

Hertzog

Rehwinkel

2020

EMBO Reports

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Cell-autonomous sensing of nucleic acids is essential for host defence against invading pathogens by inducing antiviral and inflammatory cytokines. cGAS has emerged in recent years as a non-redundant DNA sensor important for detection of many viruses and bacteria. Upon binding to DNA, cGAS synthesises the cyclic dinucleotide 2 0 3 0-cGAMP that binds to the adaptor protein STING and thereby triggers IRF3-and NFjB-dependent transcription. In addition to infection, the pathophysiology of an everincreasing number of sterile inflammatory conditions in humans involves the recognition of DNA through cGAS. Consequently, the cGAS/STING signalling axis has emerged as an attractive target for pharmacological modulation. However, the development of cGAS and STING inhibitors has just begun and a need for specific and effective compounds persists. In this review, we focus on cGAS and explore how its activation by immunostimulatory DNA is regulated by cellular mechanisms, viral immune modulators and small molecules. We further use our knowledge of cGAS modulation by cells and viruses to conceptualise potential new ways of pharmacological cGAS targeting.

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Small molecule inhibition of human cGAS reduces total cGAMP output and cytokine expression in cells

Cited by 6 publications

References 58 publications

Recognition of HIV-1 Capsid Licenses Innate Immune Response to Viral Infection

Recognition of HIV-1 Capsid Licenses Innate Immune Response to Viral Infection

Pharmacological Activation of cGAS for Cancer Immunotherapy

Regulation and inhibition of the DNA sensor cGAS

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