Design of amidobenzimidazole STING receptor agonists with systemic activity

Ramanjulu, Joshi M.; Pesiridis, G. Scott; Yang, Jing; Concha, N.O.; Singhaus, Robert R.; Zhang, S.Y.; Tran, Jean-Luc; Moore, Patrick; Lehmann, Stephanie; Eberl, H. Christian; Muelbaier, Marcel; Schneck, Jessica L.; Clemens, Jim; Adam, Michael; Mehlmann, John F.; Romano, Joseph J.; Morales, Angel; Kang, James; Leister, Lara K.; Graybill, Todd L.; Charnley, Adam K.; Ye, G.; Nevins, Neysa; Behnia, Kamelia; Wolf, Amaya I.; Kasparcova, Viera; Nurse, Kelvin; Wang, L.; Li, Y.; Klein, Michael; Hopson, Christopher B.; Guss, Jeffrey; Bantscheff, Marcus; Bergamini, Giovanna; Reilly,; Lian, Yiqian; Duffy, Kevin J.; Adams, Josetta; Foley, Kevin P.; Gough, Peter J.; Marquis, Robert W.; Smothers, James; Hoos, Axel; Bertin, John

doi:10.1038/s41586-018-0705-y

Cited by 569 publications

(517 citation statements)

References 24 publications

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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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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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“…The investigators linked two of these to create a dimeric ligand with even higher STING binding affinity. This compound demonstrated efficacy in the CT‐26 colorectal model when administered IV …”

Section: Sting Pathway Involvement In Anti‐tumor Immunity In Preclinimentioning

confidence: 99%

“…This compound demonstrated efficacy in the CT-26 colorectal model when administered IV. 121 Packaging cGAMP in liposomal nanoparticles has demonstrated improved cellular uptake and better tumor control in transplantable and genetically engineered triple negative breast cancer models as well as the B16.F10 melanoma model. 122 Another study found that liposomal formulated STING agonist can cause loss of APC viability, so the authors chose to load a STING agonist ex vivo into exosomes instead (Exosting).…”

Section: Novel Strategies For Sting Agonist Delivery By the Systemimentioning

confidence: 99%

STING pathway agonism as a cancer therapeutic

Flood

Higgs

et al. 2019

Immunological Reviews

234

183

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The fact that a subset of human cancers showed evidence for a spontaneous adaptive immune response as reflected by the T cell‐inflamed tumor microenvironment phenotype led to the search for candidate innate immune pathways that might be driving such endogenous responses. Preclinical studies indicated a major role for the host STING pathway, a cytosolic DNA sensing pathway, as a proximal event required for optimal type I interferon production, dendritic cell activation, and priming of CD8+ T cells against tumor‐associated antigens. STING agonists are therefore being developed as a novel cancer therapeutic, and a greater understanding of STING pathway regulation is leading to a broadened list of candidate immune regulatory targets. Early phase clinical trials of intratumoral STING agonists are already showing promise, alone and in combination with checkpoint blockade. Further advancement will derive from a deeper understanding of STING pathway biology as well as mechanisms of response vs resistance in individual cancer patients.

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“…Given the importance of the cGAS pathway to stimulating the innate immune system, a number of agonists and antagonists have been generated in attempts to probe the molecular underpinnings and to manipulate activity for therapeutic intervention 53,43,54,45,44 . While in vitro studies have difficulties in showing that RU.521 can inhibit recombinant human cGAS 47,44 , our data here indicate that RU.521 does inhibit human cGAS activity in cells.…”

Section: Discussionmentioning

confidence: 99%

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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Design of amidobenzimidazole STING receptor agonists with systemic activity

Cited by 569 publications

References 24 publications

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

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

STING pathway agonism as a cancer therapeutic

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

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