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

Abstract

Cyclic GMP-AMP synthase is essential for innate immunity against infection and cellular damage, serving as a sensor of DNA from pathogens or mislocalized self-DNA. Upon binding double-stranded DNA, cyclic GMP-AMP synthase synthesizes a cyclic dinucleotide that initiates an inflammatory cellular response. Mouse studies that recapitulate causative mutations in the autoimmune disease Aicardi-Goutières syndrome demonstrate that ablating the cyclic GMP-AMP synthase gene abolishes the deleterious phenotype. Here, we report the discovery of a class of cyclic GMP-AMP synthase inhibitors identified by a high-throughput screen. These compounds possess defined structure-activity relationships and we present crystal structures of cyclic GMP-AMP synthase, double-stranded DNA, and inhibitors within the enzymatic active site. We find that a chemically improved member, RU.521, is active and selective in cellular assays of cyclic GMP-AMP synthase-mediated signaling and reduces constitutive expression of interferon in macrophages from a mouse model of Aicardi-Goutières syndrome. RU.521 will be useful toward understanding the biological roles of cyclic GMP-AMP synthase and can serve as a molecular scaffold for development of future autoimmune therapies.

Authors:
 [1];  [2];  [3];  [4];  [5];  [6];  [6];  [6];  [6];  [1];  [5];  [5];  [1];  [6];  [5];  [7]; ORCiD logo [2]; ORCiD logo [1]
  1. Vanderbilt Univ. School of Medicine, Nashville, TN (United States)
  2. The Rockefeller Univ., New York, NY (United States)
  3. Memorial Sloan-Kettering Cancer Center, New York, NY (United States); Chinese Academy of Sciences (CAS), Beijing (China)
  4. Regeneron Pharmaceuticals Incorporated, Tarrytown, NY (United States); The Rockefeller Univ., New York, NY (United States)
  5. The Rockefeller Univ., New York, NY (United States); Howard Hughes Medical Inst. Lab. for RNA Molecular Biology, New York, NY (United States)
  6. Tri-Institutional Therapeutics Discovery Inst., New York, NY (United States)
  7. Memorial Sloan-Kettering Cancer Center, New York, NY (United States)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
USDOE; Vanderbilt Univ. Dept. Biochemistry; Rockefeller Univ. Robertson Therapeutic Development Funds; Cancer Research Institute Irvington Postdoctoral Fellowship; NIGMS; MSKCC
OSTI Identifier:
1430350
Grant/Contract Number:  
AC02-06CH11357; 1R35GM119569-01; NSFC31670903; 4T32GM065086-14; GM104962; P30 CA008748; P41 GM103403
Resource Type:
Accepted Manuscript
Journal Name:
Nature Communications
Additional Journal Information:
Journal Volume: 8; Journal Issue: 1; Journal ID: ISSN 2041-1723
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; High-throughput screening; Innate immunity; X-ray crystallography

Citation Formats

Vincent, Jessica, Adura, Carolina, Gao, Pu, Luz, Antonio, 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, and Ascano, Manuel. Small molecule inhibition of cGAS reduces interferon expression in primary macrophages from autoimmune mice. United States: N. p., 2017. Web. doi:10.1038/s41467-017-00833-9.
Vincent, Jessica, Adura, Carolina, Gao, Pu, Luz, Antonio, 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. Small molecule inhibition of cGAS reduces interferon expression in primary macrophages from autoimmune mice. United States. doi:10.1038/s41467-017-00833-9.
Vincent, Jessica, Adura, Carolina, Gao, Pu, Luz, Antonio, 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, and Ascano, Manuel. Fri . "Small molecule inhibition of cGAS reduces interferon expression in primary macrophages from autoimmune mice". United States. doi:10.1038/s41467-017-00833-9. https://www.osti.gov/servlets/purl/1430350.
@article{osti_1430350,
title = {Small molecule inhibition of cGAS reduces interferon expression in primary macrophages from autoimmune mice},
author = {Vincent, Jessica and Adura, Carolina and Gao, Pu and Luz, Antonio and Lama, Lodoe and Asano, Yasutomi and Okamoto, Rei and Imaeda, Toshihiro and Aida, Jumpei and Rothamel, Katherine and Gogakos, Tasos and Steinberg, Joshua and Reasoner, Seth and Aso, Kazuyoshi and Tuschl, Thomas and Patel, Dinshaw J. and Glickman, J. Fraser and Ascano, Manuel},
abstractNote = {Cyclic GMP-AMP synthase is essential for innate immunity against infection and cellular damage, serving as a sensor of DNA from pathogens or mislocalized self-DNA. Upon binding double-stranded DNA, cyclic GMP-AMP synthase synthesizes a cyclic dinucleotide that initiates an inflammatory cellular response. Mouse studies that recapitulate causative mutations in the autoimmune disease Aicardi-Goutières syndrome demonstrate that ablating the cyclic GMP-AMP synthase gene abolishes the deleterious phenotype. Here, we report the discovery of a class of cyclic GMP-AMP synthase inhibitors identified by a high-throughput screen. These compounds possess defined structure-activity relationships and we present crystal structures of cyclic GMP-AMP synthase, double-stranded DNA, and inhibitors within the enzymatic active site. We find that a chemically improved member, RU.521, is active and selective in cellular assays of cyclic GMP-AMP synthase-mediated signaling and reduces constitutive expression of interferon in macrophages from a mouse model of Aicardi-Goutières syndrome. RU.521 will be useful toward understanding the biological roles of cyclic GMP-AMP synthase and can serve as a molecular scaffold for development of future autoimmune therapies.},
doi = {10.1038/s41467-017-00833-9},
journal = {Nature Communications},
number = 1,
volume = 8,
place = {United States},
year = {2017},
month = {9}
}

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