{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T15:16:14Z","timestamp":1790090174829,"version":"4.0.1"},"posted":{"date-parts":[[2026,5,13]]},"group-title":"Cell Biology","reference-count":103,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T00:00:00Z","timestamp":1778630400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"award":["ASAP-000350"],"award-info":[{"award-number":["ASAP-000350"]}],"id":[{"id":"https:\/\/ror.org\/03zj4c476","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000001","name":"U.S. National Science Foundation","doi-asserted-by":"crossref","award":["DGE- 2236662"],"award-info":[{"award-number":["DGE- 2236662"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"crossref"}]},{"award":["1F31NS143320-01"],"award-info":[{"award-number":["1F31NS143320-01"]}],"id":[{"id":"https:\/\/ror.org\/01s5ya894","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,5,13]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Astrocytes directly influence neuronal survival and increasingly are understood to contribute to the progression of neurodegenerative diseases including Parkinson\u2019s disease (PD). Mitochondrial damage is a hallmark of PD pathology in both neurons and astrocytes. Damaged mitochondria are cleared by PINK1\/Parkin-mediated mitophagy; loss-of-function mutations in either PINK1 or Parkin are sufficient to cause PD. Neuronal mitophagy is well-studied, but far less is known about how mitochondrial dysfunction in astrocytes affects neural health. While microglial release of pro-inflammatory cytokines has been shown to induce astrocytes to mount their own inflammatory response, we hypothesize that a more direct pathway is involved, and that mitochondrial damage to astrocytes directly triggers release of proinflammatory cytokines. To address these questions, we treated primary murine cortical astrocytes with oxidative phosphorylation (OXPHOS) inhibitors antimycin A (AA) and oligomycin A (OA) and observed the PINK1-dependent accumulation of Parkin on damaged mitochondria, leading to phospho-ubiquitination of proteins in the outer mitochondrial membrane and the recruitment of the autophagy receptor SQSTM1\/p62. To identify transcriptional changes caused by mitochondrial damage and the resulting activation of mitophagic machinery, we performed bulk RNA-sequencing on astrocytes isolated from WT, PINK1\n                  <jats:sup>-\/-<\/jats:sup>\n                  , or Parkin\n                  <jats:sup>-\/-<\/jats:sup>\n                  mice treated with AA\/OA or a vehicle control. In WT astrocytes, TNF-\u03b1 signaling via NF-\u03baB was the most significantly upregulated pathway following OXPHOS inhibition. OXPHOS inhibitor treatment also stimulated p62 expression, while NF-\u03baB inhibition prevented this upregulation. Astrocytic secretion of cytokines, including TNF-\u03b1, was increased following mitochondrial damage; this secretion was dependent on NF-\u03baB activation and occurred at levels sufficient to induce mitochondrial depolarization in hippocampal neurons. Compared to WT astrocytes, PINK1\n                  <jats:sup>-\/-<\/jats:sup>\n                  astrocytes showed a significant reduction in transcriptional signatures associated with TNF-\u03b1 signaling following mitochondrial damage, while Parkin\n                  <jats:sup>-\/-<\/jats:sup>\n                  astrocytes exhibited upregulation of both IFN-\u03b3 and IFN-\u03b1 signaling. These findings indicate altered inflammatory responses to mitochondrial damage in the absence of functional PINK1 or Parkin. Finally, we analyzed scRNA-sequencing data from substantia nigra astrocytes harvested from human brain tissue from PD-positive or control samples. Distinct clusters comprised predominantly of PD-positive or control astrocytes emerged. Astrocytes in the PD-positive cluster were enriched for NF-\u03baB, IFN-\u03b1 and IFN-\u03b3 responses, consistent with the signaling observed\n                  <jats:italic>in vitro<\/jats:italic>\n                  post-OXPHOS inhibition. Together, these findings identify inflammatory signatures activated by mitochondrial damage in astrocytes, and establish this pathway as a potential contributor to neuroinflammation in PD.\n                <\/jats:p>","DOI":"10.64898\/2026.05.11.724378","type":"posted-content","created":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T21:25:14Z","timestamp":1778707514000},"source":"Crossref","is-referenced-by-count":2,"title":["PINK1\/Parkin-dependent mitophagy mediates astrocytic inflammatory responses to mitochondrial damage"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8518-9786","authenticated-orcid":false,"given":"Julia F.","family":"Riley","sequence":"first","affiliation":[{"name":"Physiology Department at the University of Pennsylvania, Philadelphia, PA 19104"},{"name":"Aligning Science Across Parkinson\u2019s (ASAP) Collaborative Research Network, Chevy Chase, MD, 20815"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-2722-6233","authenticated-orcid":false,"given":"Charity V.","family":"Robbins","sequence":"additional","affiliation":[{"name":"Physiology Department at the University of Pennsylvania, Philadelphia, PA 19104"},{"name":"First Exposure to Research in Biomedical Sciences Program at the University of Pennsylvania, Philadelphia, PA 19104"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5389-4114","authenticated-orcid":false,"given":"Erika L. 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