{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T09:54:10Z","timestamp":1785405250967,"version":"3.56.0"},"posted":{"date-parts":[[2026,6,8]]},"group-title":"Cancer Biology","reference-count":52,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,6,8]],"date-time":"2026-06-08T00:00:00Z","timestamp":1780876800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"award":["CA250506"],"award-info":[{"award-number":["CA250506"]}],"id":[{"id":"https:\/\/ror.org\/040gcmg81","id-type":"ROR","asserted-by":"publisher"}]},{"award":["CA271176"],"award-info":[{"award-number":["CA271176"]}],"id":[{"id":"https:\/\/ror.org\/040gcmg81","id-type":"ROR","asserted-by":"publisher"}]},{"award":["P50CA098131"],"award-info":[{"award-number":["P50CA098131"]}],"id":[{"id":"https:\/\/ror.org\/040gcmg81","id-type":"ROR","asserted-by":"publisher"}]},{"award":["5IK6BX005391"],"award-info":[{"award-number":["5IK6BX005391"]}],"id":[{"id":"https:\/\/ror.org\/05rsv9s98","id-type":"ROR","asserted-by":"publisher"}]},{"award":["5101BX000134"],"award-info":[{"award-number":["5101BX000134"]}],"id":[{"id":"https:\/\/ror.org\/05rsv9s98","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000090","name":"Congressionally Directed Medical Research Programs","doi-asserted-by":"crossref","award":["W81XWH2210109"],"award-info":[{"award-number":["W81XWH2210109"]}],"id":[{"id":"10.13039\/100000090","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,6,8]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  Distant metastasis is the leading cause of mortality in many cancers. Although metabolic reprogramming is recognized as a hallmark of cancer, how tumor-intrinsic metabolic enzymes regulate tumor-immune crosstalk during metastatic progression remains poorly understood. Here, using a high-throughput functional CRISPR-Cas9 screen targeting metabolic genes in an orthotopic 4T1 murine mammary carcinoma model of spontaneous lung metastasis, we identify a selective enrichment of arginase 2 (ARG2)-deficient tumor cells in metastatic lungs of immunocompetent but not RAG1-deficient mice, indicating a lymphocyte-dependent mechanism. Loss of ARG2 enhances spontaneous lung metastasis without affecting primary tumor growth. Further, metastatic outgrowth in the lung is not affected when tumor cells are injected intravenously, indicating that ARG2 regulates an early stage of the metastatic cascade. Mechanistically, ARG2 deficiency upregulates nitric oxide synthase 2 (NOS2), resulting in increased nitric oxide production, accumulation of cytosolic DNA, and activation of the cGAS-STING-NF-\u03baB pathway, leading to upregulation of inflammatory cytokines. ARG2-deficient tumors exhibit an immunosuppressive tumor microenvironment characterized by enrichment of Th17 cells and reduced anti-tumor immune populations. Functionally, Th17 cells enhance tumor cell migration\n                  <jats:italic>in vitro<\/jats:italic>\n                  and promote spontaneous lung metastasis\n                  <jats:italic>in vivo<\/jats:italic>\n                  . Genetic deletion of NOS2 attenuates cytosolic DNA accumulation, reduces STING-NF-\u03baB activation, restores anti-tumor immunity, and suppresses ARG2 deficiency-driven metastatic burden\n                  <jats:italic>in vivo<\/jats:italic>\n                  . Collectively, these findings define a tumor cell-intrinsic ARG2-NOS2 axis that regulates inflammatory signaling and the tumor microenvironment to promote metastasis, highlighting a targetable vulnerability in metastatic breast cancer.\n                <\/jats:p>","DOI":"10.64898\/2026.06.04.730112","type":"posted-content","created":{"date-parts":[[2026,6,9]],"date-time":"2026-06-09T03:55:14Z","timestamp":1780977314000},"source":"Crossref","is-referenced-by-count":0,"title":["Loss of Arginase 2 Promotes Lung Metastasis in immune-competent hosts via Nitric Oxide Synthase 2-Dependent Th17 Response"],"prefix":"10.64898","author":[{"given":"Shu-Ting","family":"Chou","sequence":"first","affiliation":[{"name":"Program in Cancer Biology, Vanderbilt University, Nashville, Tennessee"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoyong","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Medicine, Division of Rheumatology and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Yang","sequence":"additional","affiliation":[{"name":"Center for Quantitative Sciences, Vanderbilt University Medical Center, Nashville, Tennessee"},{"name":"Department of Biostatistics, Vanderbilt University Medical Center, Nashville, Tennessee"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yoonha","family":"Hwang","sequence":"additional","affiliation":[{"name":"Department of Medicine, Division of Rheumatology and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiacheng","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Computer Science, Vanderbilt University, Nashville, Tennessee"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yujie","family":"Ding","sequence":"additional","affiliation":[{"name":"Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeffrey C.","family":"Rathmell","sequence":"additional","affiliation":[{"name":"Department of Pathology, Microbiology, and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee"},{"name":"Vanderbilt-Ingram Cancer Center, Nashville, Tennessee"},{"name":"Vanderbilt Center for Immunobiology, Vanderbilt University Medical Center, Nashville, Tennessee"},{"name":"Ben May Department for Cancer Research, University of Chicago, Chicago, Illinois"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Deanna N.","family":"Edwards","sequence":"additional","affiliation":[{"name":"Department of Medicine, Division of Rheumatology and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee"},{"name":"Vanderbilt-Ingram Cancer Center, Nashville, Tennessee"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin","family":"Chen","sequence":"additional","affiliation":[{"name":"Program in Cancer Biology, Vanderbilt University, Nashville, Tennessee"},{"name":"Department of Medicine, Division of Rheumatology and Immunology, Vanderbilt University Medical Center, Nashville, Tennessee"},{"name":"Vanderbilt-Ingram Cancer Center, Nashville, Tennessee"},{"name":"Vanderbilt Institute for Infection, Immunology, and Inflammation, Nashville, Tennessee"},{"name":"Department of Cell and Developmental Biology, Vanderbilt University, Nashville, Tennessee"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026061018301401000_2026.06.04.730112v1.1","doi-asserted-by":"crossref","unstructured":"Zhang M , Deng H , Hu R , Chen F , Dong S , Zhang S , Guo W , Yang W , Chen W . Patterns and prognostic implications of distant metastasis in breast cancer based on SEER population data. Sci Rep. 15(1):26717, 2025.","DOI":"10.1038\/s41598-025-12883-x"},{"key":"2026061018301401000_2026.06.04.730112v1.2","doi-asserted-by":"publisher","DOI":"10.3322\/caac.21863"},{"key":"2026061018301401000_2026.06.04.730112v1.3","doi-asserted-by":"publisher","DOI":"10.1038\/s41568-020-00320-2"},{"key":"2026061018301401000_2026.06.04.730112v1.4","doi-asserted-by":"publisher","DOI":"10.1126\/science.aaw5473"},{"issue":"47","key":"2026061018301401000_2026.06.04.730112v1.5","doi-asserted-by":"crossref","first-page":"3447","DOI":"10.1038\/s41388-023-02868-3","article-title":"Metabolic control of cancer metastasis: role of amino acids at secondary organ sites","volume":"42","year":"2023","journal-title":"Oncogene"},{"issue":"1","key":"2026061018301401000_2026.06.04.730112v1.6","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1038\/s41420-022-01200-4","article-title":"Arginase: shedding light on the mechanisms and opportunities in cardiovascular diseases","volume":"8","year":"2022","journal-title":"Cell Death Discov"},{"key":"2026061018301401000_2026.06.04.730112v1.7","doi-asserted-by":"publisher","DOI":"10.1016\/j.biopha.2022.112840"},{"key":"2026061018301401000_2026.06.04.730112v1.8","doi-asserted-by":"publisher","DOI":"10.1126\/science.2432665"},{"key":"2026061018301401000_2026.06.04.730112v1.9","doi-asserted-by":"publisher","DOI":"10.1002\/jlb.54.2.171"},{"key":"2026061018301401000_2026.06.04.730112v1.10","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00037.2016"},{"key":"2026061018301401000_2026.06.04.730112v1.11","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-14-3443"},{"key":"2026061018301401000_2026.06.04.730112v1.12","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-18-2139"},{"issue":"6","key":"2026061018301401000_2026.06.04.730112v1.13","doi-asserted-by":"crossref","first-page":"997","DOI":"10.1016\/j.freeradbiomed.2010.06.006","article-title":"Arginase 2 and nitric oxide synthase: pathways associated with the pathogenesis of thyroid tumors","volume":"49","year":"2010","journal-title":"Free Radic Biol Med"},{"key":"2026061018301401000_2026.06.04.730112v1.14","doi-asserted-by":"crossref","first-page":"1528972","DOI":"10.3389\/fcell.2025.1528972","article-title":"Arginase-II promotes melanoma and lung cancer cell growth by regulating Sirt3-mtROS axis","volume":"13","year":"2025","journal-title":"Front Cell Dev Biol"},{"issue":"2","key":"2026061018301401000_2026.06.04.730112v1.15","doi-asserted-by":"crossref","first-page":"e71567","DOI":"10.1002\/cam4.71567","article-title":"Arginase 2 promotes colorectal cancer metastasis via PI3K\/AKT pathway activation and regulates tumor immune infiltration","volume":"15","year":"2026","journal-title":"Cancer Med"},{"issue":"14","key":"2026061018301401000_2026.06.04.730112v1.16","doi-asserted-by":"crossref","first-page":"1561","DOI":"10.1002\/pros.20816","article-title":"Disruption of arginase II alters prostate tumor formation in TRAMP mice","volume":"68","year":"2008","journal-title":"Prostate"},{"key":"2026061018301401000_2026.06.04.730112v1.17","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2018.04.009"},{"issue":"16","key":"2026061018301401000_2026.06.04.730112v1.18","doi-asserted-by":"crossref","first-page":"e193091","DOI":"10.1172\/jci.insight.193091","article-title":"Elevated tumor NOS2\/COX2 promotes immunosuppressive phenotypes associated with poor survival in ER\u2013 breast cancer","volume":"10","year":"2025","journal-title":"JCI Insight"},{"key":"2026061018301401000_2026.06.04.730112v1.19","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1709119114"},{"key":"2026061018301401000_2026.06.04.730112v1.20","doi-asserted-by":"publisher","DOI":"10.1126\/science.1232458"},{"key":"2026061018301401000_2026.06.04.730112v1.21","doi-asserted-by":"publisher","DOI":"10.1126\/science.1229963"},{"key":"2026061018301401000_2026.06.04.730112v1.22","doi-asserted-by":"publisher","DOI":"10.1038\/nature10429"},{"key":"2026061018301401000_2026.06.04.730112v1.23","doi-asserted-by":"publisher","DOI":"10.1038\/nature07317"},{"key":"2026061018301401000_2026.06.04.730112v1.24","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.162356399"},{"key":"2026061018301401000_2026.06.04.730112v1.25","doi-asserted-by":"publisher","DOI":"10.1053\/gast.2001.20875"},{"issue":"3","key":"2026061018301401000_2026.06.04.730112v1.26","doi-asserted-by":"crossref","first-page":"G626","DOI":"10.1152\/ajpgi.2001.281.3.G626","article-title":"Nitric oxide in gastrointestinal epithelial cell carcinogenesis: linking inflammation to oncogenesis","volume":"281","year":"2001","journal-title":"Am J Physiol Gastrointest Liver Physiol"},{"key":"2026061018301401000_2026.06.04.730112v1.27","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20071397"},{"key":"2026061018301401000_2026.06.04.730112v1.28","doi-asserted-by":"publisher","DOI":"10.1038\/nature04753"},{"key":"2026061018301401000_2026.06.04.730112v1.29","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2006.01.001"},{"issue":"5","key":"2026061018301401000_2026.06.04.730112v1.30","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1038\/s41577-025-01250-6","article-title":"Contextualizing TH17 cells in cancer","volume":"26","year":"2026","journal-title":"Nat Rev Immunol"},{"key":"2026061018301401000_2026.06.04.730112v1.31","doi-asserted-by":"publisher","DOI":"10.1172\/JCI124037"},{"key":"2026061018301401000_2026.06.04.730112v1.32","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2023.1094823"},{"key":"2026061018301401000_2026.06.04.730112v1.33","doi-asserted-by":"publisher","DOI":"10.1038\/onc.2012.105"},{"key":"2026061018301401000_2026.06.04.730112v1.34","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0079242"},{"key":"2026061018301401000_2026.06.04.730112v1.35","doi-asserted-by":"publisher","DOI":"10.1242\/dmm.017830"},{"key":"2026061018301401000_2026.06.04.730112v1.36","doi-asserted-by":"publisher","DOI":"10.4161\/15548627.2014.981789"},{"key":"2026061018301401000_2026.06.04.730112v1.37","doi-asserted-by":"publisher","DOI":"10.3978\/j.issn.2072-1439.2013.06.17"},{"key":"2026061018301401000_2026.06.04.730112v1.38","doi-asserted-by":"publisher","DOI":"10.1016\/j.immuni.2010.07.001"},{"key":"2026061018301401000_2026.06.04.730112v1.39","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamcr.2017.02.005"},{"key":"2026061018301401000_2026.06.04.730112v1.40","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2020.01.108"},{"key":"2026061018301401000_2026.06.04.730112v1.41","doi-asserted-by":"publisher","DOI":"10.1038\/s41568-018-0050-3"},{"key":"2026061018301401000_2026.06.04.730112v1.42","doi-asserted-by":"crossref","first-page":"770","DOI":"10.1186\/s12885-015-1808-6","article-title":"The NF-\u03baB p65 and p50 homodimer cooperate with IRF8 to activate iNOS transcription","volume":"15","year":"2015","journal-title":"BMC Cancer"},{"key":"2026061018301401000_2026.06.04.730112v1.43","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.181.4.2799"},{"key":"2026061018301401000_2026.06.04.730112v1.44","doi-asserted-by":"publisher","DOI":"10.1038\/s41588-021-00911-1"},{"key":"2026061018301401000_2026.06.04.730112v1.45","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccell.2023.01.009"},{"issue":"2","key":"2026061018301401000_2026.06.04.730112v1.46","doi-asserted-by":"crossref","first-page":"366","DOI":"10.1038\/s12276-026-01650-1","article-title":"Metabolic crosstalk among cancer-associated fibroblasts, adipocytes and immune cells as an immunosuppressive tumor microenvironment driver","volume":"58","year":"2026","journal-title":"Exp Mol Med"},{"issue":"8","key":"2026061018301401000_2026.06.04.730112v1.47","doi-asserted-by":"crossref","first-page":"2019","DOI":"10.1016\/j.immuni.2025.07.007","article-title":"Polyamines regulate adaptive antitumor immunity by functional specialization of regulatory T cells","volume":"58","year":"2025","journal-title":"Immunity"},{"key":"2026061018301401000_2026.06.04.730112v1.48","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2305245120"},{"key":"2026061018301401000_2026.06.04.730112v1.49","doi-asserted-by":"publisher","DOI":"10.1126\/science.abj3510"},{"issue":"1","key":"2026061018301401000_2026.06.04.730112v1.50","doi-asserted-by":"crossref","first-page":"143","DOI":"10.1016\/j.immuni.2022.12.010","article-title":"T cell-derived interleukin-22 drives the expression of CD155 by cancer cells to suppress NK cell function and promote metastasis","volume":"56","year":"2023","journal-title":"Immunity"},{"issue":"9","key":"2026061018301401000_2026.06.04.730112v1.51","doi-asserted-by":"crossref","first-page":"e187531","DOI":"10.1172\/jci.insight.187531","article-title":"Increased fatty acid delivery by tumor endothelium promotes metastatic outgrowth","volume":"10","year":"2025","journal-title":"JCI Insight"},{"key":"2026061018301401000_2026.06.04.730112v1.52","doi-asserted-by":"publisher","DOI":"10.1126\/science.1260419"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.06.04.730112","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,11]],"date-time":"2026-06-11T01:30:27Z","timestamp":1781141427000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.06.04.730112"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,6,8]]},"references-count":52,"URL":"https:\/\/doi.org\/10.64898\/2026.06.04.730112","relation":{},"subject":[],"published":{"date-parts":[[2026,6,8]]},"subtype":"preprint"}}