{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T19:04:26Z","timestamp":1790881466161,"version":"4.1.0"},"posted":{"date-parts":[[2026,9,29]]},"group-title":"Cell Biology","reference-count":25,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T00:00:00Z","timestamp":1790640000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"award":["P01HL185367"],"award-info":[{"award-number":["P01HL185367"]}],"id":[{"id":"https:\/\/ror.org\/012pb6c26","id-type":"ROR","asserted-by":"publisher"}]},{"award":["T32HD007499"],"award-info":[{"award-number":["T32HD007499"]}],"id":[{"id":"https:\/\/ror.org\/012pb6c26","id-type":"ROR","asserted-by":"publisher"}]},{"award":["Dean Scholar"],"award-info":[{"award-number":["Dean Scholar"]}],"id":[{"id":"https:\/\/ror.org\/01yc7t268","id-type":"ROR","asserted-by":"publisher"}]},{"award":["Elizabeth McDonnell Endowed Chair in Pediatric Hematology Oncology"],"award-info":[{"award-number":["Elizabeth McDonnell Endowed Chair in Pediatric Hematology Oncology"]}],"id":[{"id":"https:\/\/ror.org\/01yc7t268","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,29]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>TNF\u03b1 is the primary age-related cytokine responsible for hyperreactive platelet formation. In mouse models, increased TNF\u03b1 induced abnormal megakaryocyte development and platelet activity. Here, we extend these findings to demonstrate that TNF\u03b1 drives this thrombotic phenotype through expansion of the hematopoietic stem and progenitor cell (HSPCs) compartment. Using HSPC-specific TNF\u03b1 receptor labeling and single cell RNA sequencing, we found that TNF\u03b1 receptors are absent from megakaryocytes and their progenitors (MkPs) indicating that TNF\u03b1 does not directly act on these cells. Chronic TNF\u03b1 exposure expanded HSPCs in the bone marrow and extramedullary tissues \u2013 and these expanded HSPCs retained functional repopulation capacity. Using species-specificity TNF\u03b1 receptor activation, we further demonstrate that TNF\u03b1R1 signaling is sufficient to induce platelet hyperreactivity independent of HSPC expansion. Unlike emergency hematopoiesis, chronic TNF\u03b1 promoted megakaryopoiesis through the canonical hematopoietic hierarchy as demonstrated by lineage-tracing studies. Mechanistically, chronic TNF\u03b1 induced a distinct transcriptional program in single cell RNA sequencing of HSPCs and megakaryocytes. Together, these findings establish that chronic TNF\u03b1 promotes hyperreactive platelet formation not through direct effects on megakaryocytes or MkPs but by expanding and transcriptionally reprogramming HSPCs, thereby imprinting a TNF\u03b1 dependent program that persists through megakaryopoiesis and ultimately produces hyperreactive platelets.<\/jats:p>\n                <jats:sec>\n                  <jats:title>Key Points<\/jats:title>\n                  <jats:list list-type=\"order\">\n                    <jats:list-item>\n                      <jats:p>Chronic TNF\u03b1 exposure drives HSC expansion and myeloid biasing of progenitors.<\/jats:p>\n                    <\/jats:list-item>\n                    <jats:list-item>\n                      <jats:p>TNF\u03b1 regulates platelet hyperreactivity by signaling through HSPCs, not Mks.<\/jats:p>\n                    <\/jats:list-item>\n                  <\/jats:list>\n                <\/jats:sec>","DOI":"10.64898\/2026.09.28.755049","type":"posted-content","created":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T15:10:22Z","timestamp":1790694622000},"source":"Crossref","is-referenced-by-count":0,"title":["TNF\u03b1 drives platelet hyperreactivity and thromboinflammation through regulation of hematopoietic stem and progenitor cells"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9103-3224","authenticated-orcid":false,"given":"Joshua I","family":"Siner","sequence":"first","affiliation":[{"name":"Division of Hematology, Washington University School of Medicine, Saint Louis, MO"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7634-9562","authenticated-orcid":false,"given":"Molly","family":"Brakhane","sequence":"additional","affiliation":[{"name":"Division of Hematology, Washington University School of Medicine, Saint Louis, 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