Small molecule inhibition of cGAS reduces interferon expression in primary macrophages from autoimmune mice

Vincent, Jessica; Adura, Carolina; Gao, Pu; Luz, Antonio N.; Lama, Lodoe; Asano, Yasutomi; Okamoto, Rei; Imaeda, Toshihiro; Aida, Jumpei; Rothamel, Katherine; Gogakos, Tasos; Steinberg, Joshua; Reasoner, Seth; Aso, Kazuyoshi; Tuschl, Thomas; Patel, Dinshaw J.; Glickman, J. Fraser; Ascano, Manuel

doi:10.1038/s41467-017-00833-9

Cited by 223 publications

(197 citation statements)

References 60 publications

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“…Using concentrations spanning the range from 0.001 µM to 100 µM, the IC50 was determined to be ~ 0.8 µM in THP-1 cells. These findings are in line with our previous assessment of RU.521 potency against mouse cGAS (m-cGAS), which was measured to be 0.7 µM in murine RAW 264.7 cell 46 . These results indicate that RU.521 is capable of inhibiting DNA-and h-cGAS-dependent immune activation.…”

Section: For Determination Of Ec50supporting

confidence: 92%

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

Wiser

Kim

Vincent

et al. 2020

Preprint

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ABSTRACTThe cGAS-STING pathway is a major mechanism that mammalian cells utilize to detect cytoplasmic dsDNA from incoming viruses, bacteria, or self. CYCLIC GMP-AMP SYNTHASE (cGAS) is the sensor protein that directly binds dsDNAs. cGAS synthesizes cyclic GMP-AMP (cGAMP), which binds to the adaptor STIMULATOR OF INTERFERON GENES (STING), activating an INTERFERON REGULATORY FACTOR 3 (IRF3)-mediated immune response. Constitutive activation can result in interferonopathies such as Aicardi-Goutieres Syndrome (AGS) or other lupus-like autoimmune disorders. While inhibitors targeting mouse or human cGAS have been reported, the identification of a small molecule that targets both homologs of cGAS has been challenging. Here, we show that RU.521 is capable of potently and selectively inhibiting mouse and human cGAS in cell lines and human primary cells. This inhibitory activity requires the presence of cGAS, but it cannot suppress an immune response in cells activated by RNA, Toll-like receptor ligands, cGAMP, or recombinant interferon. Importantly, when RU.521 is applied to cells, the production of dsDNA-induced intracellular cGAMP is suppressed in a dose-dependent manner. Our work validates the use of RU.521 for probing DNA-induced innate immune responses and underscores its potential as an ideal scaffold towards pre-clinical development, given its potency against human and mouse cGAS.

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Section: For Determination Of Ec50supporting

confidence: 92%

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

Wiser

Kim

Vincent

et al. 2020

Preprint

Self Cite

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“…The cGAS-STING antiviral response has become the subject of intense development in the pharmaceutical industry, including efforts to develop inhibitors of cGAS and STING to treat human autoimmune diseases (35)(36)(37), as well as agonists of STING to improve immune responses to tumors (38)(39)(40). Our discovery of a second DNA-activated antiviral response in human cells has important implications for these efforts.…”

Section: Discussionmentioning

confidence: 99%

Human DNA-PK activates a STING-independent DNA sensing pathway

Burleigh

Maltbaek

Cambier

et al. 2019

Preprint

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Detection of intracellular DNA by the cGAS-STING pathway activates a type I interferon-mediated innate immune response that protects from virus infection and can be harnessed to promote anti-tumor immunity. Whether there are additional DNA sensing pathways, and how such pathways might function, remains controversial. We show here that humans -but not mice -have a second, potent, STINGindependent DNA sensing pathway that is blocked by the E1A viral oncogene of human adenovirus 5.We identify human DNA-PK as the sensor of this pathway and demonstrate that DNA-PK kinase activity drives a robust and broad antiviral response. We discover that the heat shock protein HSPA8/HSC70 is a unique target of DNA-PK. Finally, we demonstrate that detection of foreign DNA and DNA damage trigger distinct modalities of DNA-PK activity. These findings reveal the existence, sensor, unique target, and viral antagonists of a STING-independent DNA sensing pathway (SIDSP) in human cells. IntroductionThe cGAS-STING DNA sensing pathway has emerged as a key component of the innate immune response that is important for antiviral immunity (1), contributes to specific autoimmune diseases (2), and mediates important aspects of antitumor immunity (3). cGAS binds to double-stranded DNA and catalyzes the formation of cyclic GMP-AMP (4, 5), a diffusible cyclic dinucleotide that activates the endoplasmic adaptor protein STING (6). Activated STING then serves as a platform for the inducible recruitment of the TBK1 kinase, which phosphorylates and activates the transcription factor IRF3, leading to the induction of the type I interferon mediated antiviral response (7).Nearly all studies on the cGAS-STING pathway involve the use of mice and mouse cells.Knockouts of cGAS (Mb21d1) and STING (Tmem173) have clearly demonstrated that both are essential for the transcriptional and cell biological responses to foreign intracellular DNA, and that they mediate the pathology of specific autoimmune diseases (2,8,9). However, numerous additional sensors of intracellular DNA have been proposed, all of which are thought to act upstream of STING (10). Whether STING-independent DNA sensing exists is currently unknown.Here, we report the unexpected finding that the E1A oncogene of human adenovirus 5 blocks two distinct DNA sensing pathways in human cells: the well-known cGAS-STING pathway (11), and a second, STING-independent DNA sensing pathway (SIDSP). We identify the DNA damage response protein DNA-PK as the sensor of the SIDSP, along with the heat shock protein HSPA8 as a unique SIDSP target. We show that the SIDSP is potently activated in human and primate cells, but it is weak or absent from mouse cells. Our findings demonstrate that human cells have a second DNA sensing pathway, with implications for host defense, autoimmunity, and anti-tumor immunity. Results Human adenovirus 5 E1A blocks two DNA sensing pathways in human cellsWe previously demonstrated that the viral oncogenes of the DNA tumor viruses are potent antagonists of the cGAS-STING DNA sensing pathway ...

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“…In autoimmunity of Aicardi‐Goutières syndrome, it has been exposed that RU.521 is dynamic and elective in cellular immune functionality of cGAS‐mediated immune signaling and decreases induction of IFN in macrophages in a mouse model. RU.521 can assist as a constituent for the progress of prospective autoimmune disease therapy 147. A new investigation discloses that acetylation adds to the regulation of cGAS activity and delivers a potential therapy for handling DNA‐mediated autoimmune diseases 148…”

Section: Targeting Innate Immune Agents For Immunotherapymentioning

confidence: 99%

Regulation of cGAS‐Mediated Immune Responses and Immunotherapy

et al. 2020

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Early detection of infectious nucleic acids released from invading pathogens by the innate immune system is critical for immune defense. Detection of these nucleic acids by host immune sensors and regulation of DNA sensing pathways have been significant interests in the past years. Here, current understandings of evolutionarily conserved DNA sensing cyclic GMP‐AMP (cGAMP) synthase (cGAS) are highlighted. Precise activation and tight regulation of cGAS are vital in appropriate innate immune responses, senescence, tumorigenesis and immunotherapy, and autoimmunity. Hence, substantial insights into cytosolic DNA sensing and immunotherapy of indispensable cytosolic sensors have been detailed to extend limited knowledge available thus far. This Review offers a critical, in‐depth understanding of cGAS regulation, cytosolic DNA sensing, and currently established therapeutic approaches of essential cytosolic immune agents for improved human health.

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Small molecule inhibition of cGAS reduces interferon expression in primary macrophages from autoimmune mice

Cited by 223 publications

References 60 publications

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

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

Human DNA-PK activates a STING-independent DNA sensing pathway

Regulation of cGAS‐Mediated Immune Responses and Immunotherapy

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