{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,10]],"date-time":"2026-09-10T01:06:39Z","timestamp":1789002399601,"version":"build-2803163510"},"posted":{"date-parts":[[2026,9,7]]},"group-title":"Cell Biology","reference-count":39,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T00:00:00Z","timestamp":1788739200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health grants","doi-asserted-by":"crossref","award":["R01 AG077732, R01 AI161022, R01 AR082192, R01 AR081270, NIH P30 AR074992"],"award-info":[{"award-number":["R01 AG077732, R01 AI161022, R01 AR082192, R01 AR081270, NIH P30 AR074992"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,7]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  The NLRP3 D301N substitution, the murine ortholog of the human D303N variant, causes NLRP3 constitutive activation (NLRP3\n                  <jats:sup>CA<\/jats:sup>\n                  ) and inflammasome assembly. Here, we show that priming signals induced by lipopolysaccharide (LPS) are sufficient to trigger GSDMD-dependent pyroptosis in NLRP3\n                  <jats:sup>CA<\/jats:sup>\n                  -expressing bone marrow-derived macrophages (BMDMs), but not in wild- type (NLRP3\n                  <jats:sup>WT<\/jats:sup>\n                  ) cells. NLRP3\n                  <jats:sup>CA<\/jats:sup>\n                  mice exhibit elevated IL-1\u03b2 secretion and LDH release in bone marrow supernatants under both basal and LPS-challenged conditions. Other cytokines (e.g., TNF-\u03b1) increased comparably between genotypes, indicating that NLRP3\n                  <jats:sup>CA<\/jats:sup>\n                  specifically amplifies inflammasome-dependent responses. Conditioned medium (CM) from LPS-treated NLRP3\n                  <jats:sup>CA<\/jats:sup>\n                  BMDMs significantly enhanced osteoclast (OC) differentiation\n                  <jats:italic>in vivo<\/jats:italic>\n                  and\n                  <jats:italic>in vitro<\/jats:italic>\n                  compared to NLRP3\n                  <jats:sup>WT<\/jats:sup>\n                  CM, as did bone marrow supernatants from NLRP3\n                  <jats:sup>CA<\/jats:sup>\n                  mice. This osteoclastogenic activity was largely independent of IL-1\u03b2, as demonstrated by experiments using IL-1\u03b2-deficient NLRP3\n                  <jats:sup>CA<\/jats:sup>\n                  BMDMs, IL-1\u03b2-neutralizing antibodies, and genetic deletion of the IL-1 receptor. The osteoclastogenic factors were primarily soluble proteins, as heat inactivation and proteinase K digestion markedly reduced OC formation. Extracellular vesicles (EVs) from NLRP3\n                  <jats:sup>CA<\/jats:sup>\n                  CM modestly promoted OC differentiation, but EV-depleted supernatants retained full activity, indicating that the primary mediators are soluble proteins not carried within EVs. Together, these findings establish NLRP3\n                  <jats:sup>CA<\/jats:sup>\n                  as a well-controlled model for studying pyroptosis and demonstrate that pyroptotic cells release soluble proteins that drive osteoclastogenesis independently of IL- 1\u03b2.\n                <\/jats:p>","DOI":"10.64898\/2026.09.03.748049","type":"posted-content","created":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T07:15:16Z","timestamp":1788851716000},"source":"Crossref","is-referenced-by-count":0,"title":["Pyroptotic cell-derived factors promote osteoclast differentiation"],"prefix":"10.64898","author":[{"given":"Yongjia","family":"Li","sequence":"first","affiliation":[{"name":"Division of Bone and Mineral Diseases, Washington University School of Medicine; St. Louis, MO 63110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chun","family":"Wang","sequence":"additional","affiliation":[{"name":"Division of Bone and Mineral Diseases, Washington University School of Medicine; St. Louis, MO 63110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Canxin","family":"Xu","sequence":"additional","affiliation":[{"name":"Aclaris Therapeutics Inc., St. Louis, MO 63108, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wei","family":"Zou","sequence":"additional","affiliation":[{"name":"Division of Bone and Mineral Diseases, Washington University School of Medicine; St. Louis, MO 63110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Khushpreet","family":"Kaur","sequence":"additional","affiliation":[{"name":"Division of Bone and Mineral Diseases, Washington University School of Medicine; St. Louis, MO 63110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ria","family":"Rohatgi","sequence":"additional","affiliation":[{"name":"Division of Bone and Mineral Diseases, Washington University School of Medicine; St. Louis, MO 63110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nitin","family":"Pokhrel","sequence":"additional","affiliation":[{"name":"Division of Bone and Mineral Diseases, Washington University School of Medicine; St. Louis, MO 63110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nidhi","family":"Rohatgi","sequence":"additional","affiliation":[{"name":"Division of Bone and Mineral Diseases, Washington University School of Medicine; St. Louis, MO 63110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yousef","family":"Abu-Amer","sequence":"additional","affiliation":[{"name":"Department of Orthopedic Surgery, Washington University School of Medicine; St. Louis, MO 63110, USA"},{"name":"Shriners Hospital for Children; St. Louis, MO, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriel","family":"Mbalaviele","sequence":"additional","affiliation":[{"name":"Division of Bone and Mineral Diseases, Washington University School of Medicine; St. Louis, MO 63110, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026090917151633000_2026.09.03.748049v1.1","doi-asserted-by":"crossref","first-page":"48","DOI":"10.1038\/s41413-022-00219-8","article-title":"Bone remodeling: an operational process ensuring survival and bone mechanical competence","volume":"10","year":"2022","journal-title":"Bone Res"},{"key":"2026090917151633000_2026.09.03.748049v1.2","doi-asserted-by":"publisher","DOI":"10.1146\/annurev-physiol-021119-034425"},{"key":"2026090917151633000_2026.09.03.748049v1.3","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1038\/s41421-025-00853-6","article-title":"Cell signaling and transcriptional regulation of osteoclast lineage commitment, differentiation, bone resorption and diseases","volume":"12","year":"2026","journal-title":"Cell Discov"},{"key":"2026090917151633000_2026.09.03.748049v1.4","doi-asserted-by":"crossref","first-page":"1639430","DOI":"10.3389\/fcell.2025.1639430","article-title":"Critical signaling pathways in osteoclast differentiation and bone resorption: mechanisms and therapeutic implications for periprosthetic osteolysis","volume":"13","year":"2025","journal-title":"Front. 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