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      Radiation causes tissue damage by dysregulating inflammasome–gasdermin D signaling in both host and transplanted cells

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          Abstract

          Radiotherapy is a commonly used conditioning regimen for bone marrow transplantation (BMT). Cytotoxicity limits the use of this life-saving therapy, but the underlying mechanisms remain poorly defined. Here, we use the syngeneic mouse BMT model to test the hypothesis that lethal radiation damages tissues, thereby unleashing signals that indiscriminately activate the inflammasome pathways in host and transplanted cells. We find that a clinically relevant high dose of radiation causes severe damage to bones and the spleen through mechanisms involving the NLRP3 and AIM2 inflammasomes but not the NLRC4 inflammasome. Downstream, we demonstrate that gasdermin D (GSDMD), the common effector of the inflammasomes, is also activated by radiation. Remarkably, protection against the injury induced by deadly ionizing radiation occurs only when NLRP3, AIM2, or GSDMD is lost simultaneously in both the donor and host cell compartments. Thus, this study reveals a continuum of the actions of lethal radiation relayed by the inflammasome-GSDMD axis, initially affecting recipient cells and ultimately harming transplanted cells as they grow in the severely injured and toxic environment. This study also suggests that therapeutic targeting of inflammasome-GSDMD signaling has the potential to prevent the collateral effects of intense radiation regimens.

          Abstract

          Radiotherapy is a commonly used conditioning regimen for bone marrow transplantation; cytotoxicity limits the use of this life-saving therapy, but the underlying mechanisms remain poorly defined. This study reveals a continuum of the effects of radiation, relayed by the inflammasome–gasdermin D axis, initially affecting host cells and, ultimately, harming transplanted cells.

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          Most cited references53

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          Chemotherapy drugs induce pyroptosis through caspase-3 cleavage of a Gasdermin

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            Gasdermin E suppresses tumor growth by activating anti-tumor immunity

            Cleavage of the gasdermins to produce a pore-forming N-terminal fragment causes inflammatory death (pyroptosis) 1 . Caspase-3 cleaves gasdermin E (GSDME, also known as DFNA5), mutated in familial aging-related hearing loss 2 , which converts noninflammatory apoptosis to pyroptosis in GSDME-expressing cells 3–5 . GSDME expression is suppressed in many cancers and reduced GSDME is associated with decreased breast cancer survival 2,6 , suggesting GSDME might be a tumor suppressor. Here we show reduced GSDME function of 20 of 22 tested cancer-associated mutations. Gsdme knockout in GSDME-expressing tumors enhances, while ectopic expression in Gsdme-repressed tumors inhibits, tumor growth. Tumor suppression is mediated by cytotoxic lymphocyte killing since it is abrogated in perforin-deficient or killer lymphocyte-depleted mice. GSDME expression enhances tumor-associated macrophage phagocytosis and the number and functions of tumor-infiltrating NK and CD8+ T lymphocytes. Killer cell granzyme B also activates caspase-independent pyroptosis in target cells by directly cleaving GSDME at the same site as caspase-3. Non-cleavable or pore-defective GSDME are not tumor suppressive. Thus, tumor GSDME is a tumor suppressor by activating pyroptosis, which enhances anti-tumor immunity.
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              The Pore-Forming Protein Gasdermin D Regulates Interleukin-1 Secretion from Living Macrophages

              The interleukin-1 (IL-1) family cytokines are cytosolic proteins that exhibit inflammatory activity upon release into the extracellular space. These factors are released following various cell death processes, with pyroptosis being a common mechanism. Recently, it was recognized that phagocytes can achieve a state of hyperactivation, which is defined by their ability to secrete IL-1 while retaining viability, yet it is unclear how IL-1 can be secreted from living cells. Herein, we report that the pyroptosis regulator gasdermin D (GSDMD) was necessary for IL-1β secretion from living macrophages that have been exposed to inflammasome activators, such as bacteria and their products or host-derived oxidized lipids. Cell- and liposome-based assays demonstrated that GSDMD pores were required for IL-1β transport across an intact lipid bilayer. These findings identify a non-pyroptotic function for GSDMD, and raise the possibility that GSDMD pores represent conduits for the secretion of cytosolic cytokines under conditions of cell hyperactivation.
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                Author and article information

                Contributors
                Role: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: Writing – review & editing
                Role: Formal analysisRole: Funding acquisitionRole: Methodology
                Role: Data curationRole: Formal analysisRole: MethodologyRole: Writing – review & editing
                Role: Formal analysisRole: Methodology
                Role: Data curationRole: Formal analysisRole: Methodology
                Role: Data curationRole: Formal analysisRole: Methodology
                Role: Data curationRole: Formal analysisRole: Methodology
                Role: ResourcesRole: SupervisionRole: Writing – review & editing
                Role: MethodologyRole: ResourcesRole: Writing – review & editing
                Role: MethodologyRole: ResourcesRole: Writing – review & editing
                Role: MethodologyRole: ResourcesRole: Writing – review & editing
                Role: Formal analysisRole: Funding acquisitionRole: InvestigationRole: MethodologyRole: Project administrationRole: ResourcesRole: SupervisionRole: Writing – original draftRole: Writing – review & editing
                Role: Academic Editor
                Journal
                PLoS Biol
                PLoS Biol
                plos
                plosbiol
                PLoS Biology
                Public Library of Science (San Francisco, CA USA )
                1544-9173
                1545-7885
                6 August 2020
                August 2020
                6 August 2020
                : 18
                : 8
                : e3000807
                Affiliations
                [1 ] Division of Bone and Mineral Diseases, Washington University School of Medicine, St. Louis, Missouri, United Sates of America
                [2 ] Division of Oncology, Washington University School of Medicine, St. Louis, Missouri, United Sates of America
                [3 ] Department of Spine Surgery, Honghui Hospital, Xi'an Jiaotong University, Xi'an, Shaanxi, China
                [4 ] Aclaris Therapeutics, Inc., St. Louis, Missouri, United Sates of America
                [5 ] Department of Orthopedic Surgery, Washington University School of Medicine, St. Louis, Missouri, United Sates of America
                [6 ] Shriners Hospital for Children, St. Louis, Missouri, United Sates of America
                National Institute of Biological Sciences, CHINA
                Author notes

                I have read the journal's policy and the authors of this manuscript have the following competing interests: GM is a consultant for Aclaris Therapeutics. KLK and JBM are employees of Aclaris Therapeutics.

                Author information
                http://orcid.org/0000-0001-7101-5582
                http://orcid.org/0000-0002-5890-5086
                http://orcid.org/0000-0003-4660-0952
                Article
                PBIOLOGY-D-20-00765
                10.1371/journal.pbio.3000807
                7446913
                32760056
                61e9acb2-5040-42da-9274-c8b162635449
                © 2020 Xiao et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 25 March 2020
                : 20 July 2020
                Page count
                Figures: 4, Tables: 0, Pages: 19
                Funding
                Funded by: funder-id http://dx.doi.org/10.13039/100000069, National Institute of Arthritis and Musculoskeletal and Skin Diseases;
                Award ID: AR068972
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100000069, National Institute of Arthritis and Musculoskeletal and Skin Diseases;
                Award ID: AR076758
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100000069, National Institute of Arthritis and Musculoskeletal and Skin Diseases;
                Award ID: AR049192
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100000069, National Institute of Arthritis and Musculoskeletal and Skin Diseases;
                Award ID: AR054326
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100000069, National Institute of Arthritis and Musculoskeletal and Skin Diseases;
                Award ID: AR072623
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100011781, Shriners Hospitals for Children;
                Award ID: 85160
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100000069, National Institute of Arthritis and Musculoskeletal and Skin Diseases;
                Award ID: AR052705
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100000069, National Institute of Arthritis and Musculoskeletal and Skin Diseases;
                Award ID: AR073507
                Award Recipient :
                Funded by: funder-id http://dx.doi.org/10.13039/100011781, Shriners Hospitals for Children;
                Award ID: 85117
                Award Recipient :
                This work was supported by NIH/NIAMS AR068972 and AR076758 grants to GM. DJV was supported by NIH grants AR052705 and AR073507 and grant 85117 from Shriners Hospitals for Children. YA-A was supported by NIH grants AR049192, AR054326, AR072623 and by grant #85160 from the Shriners Hospital for Children. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Short Reports
                Biology and Life Sciences
                Immunology
                Immune System Proteins
                Inflammasomes
                Medicine and Health Sciences
                Immunology
                Immune System Proteins
                Inflammasomes
                Biology and Life Sciences
                Biochemistry
                Proteins
                Immune System Proteins
                Inflammasomes
                Medicine and Health Sciences
                Surgical and Invasive Medical Procedures
                Blood and Lymphatic System Procedures
                Bone Marrow Transplantation
                Medicine and Health Sciences
                Surgical and Invasive Medical Procedures
                Transplantation
                Bone Marrow Transplantation
                Biology and Life Sciences
                Cell Biology
                Cellular Types
                Animal Cells
                Bone Marrow Cells
                Biology and Life Sciences
                Physiology
                Immune Physiology
                Spleen
                Biology and Life Sciences
                Anatomy
                Musculoskeletal System
                Skeleton
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                Medicine and Health Sciences
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                Radiation Therapy
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                Clinical Medicine
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                Research and Analysis Methods
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                Biology and Life Sciences
                Neuroscience
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                Medicine and Health Sciences
                Diagnostic Medicine
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                Diagnostic Radiology
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                Medicine and Health Sciences
                Radiology and Imaging
                Diagnostic Radiology
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                Research and Analysis Methods
                Animal Studies
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                Research and Analysis Methods
                Animal Studies
                Experimental Organism Systems
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                Custom metadata
                vor-update-to-uncorrected-proof
                2020-08-18
                All relevant data are within the paper and its Supporting Information files.

                Life sciences
                Life sciences

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