{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,5,14]],"date-time":"2025-05-14T02:07:10Z","timestamp":1747188430952,"version":"3.40.5"},"reference-count":77,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2023,9,18]],"date-time":"2023-09-18T00:00:00Z","timestamp":1694995200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000054","name":"National Cancer Institute","doi-asserted-by":"publisher","award":["R01CA190170","R00CA245187"],"award-info":[{"award-number":["R01CA190170","R00CA245187"]}],"id":[{"id":"10.13039\/100000054","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Vet Comparative Oncology"],"published-print":{"date-parts":[[2023,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Pharmacologic inhibition of autophagy can be achieved using lysosomotropic agents such as hydroxychloroquine (HCQ) that interfere with fusion of the autophagosome to the lysosome thus preventing completion of the recycling process. The goal of the present study is to determine the sensitivity of eight canine (cOSA) and four human (hOSA) osteosarcoma tumour cell lines to antiproliferative and cytotoxic effects of lysosomal autophagy inhibitors, and to compare these results to the autophagy\u2010dependence measured using a CRISPR\/Cas9 live\u2010cell imaging assay in OSA and other tumour cell lines. Antiproliferative and cytotoxic response to HCQ and Lys05 was determined using live cell imaging and YOYO\u20101 staining. CRISPR\/Cas9 live cell imaging screen was done using species specific guide RNA's and transfection of reagents into cells. Response to autophagy core genes was compared to response to an essential (PCNA) and non\u2010essential (FOXO3A) gene. cOSA and hOSA cell lines showed similar antiproliferative and cytotoxic responses to HCQ and Lys05 with median lethal dose (<jats:italic>D<\/jats:italic><jats:sub>m<\/jats:sub>) values ranging from 4.6\u201315.8\u2009\u03bcM and 2.1\u20135.1\u2009\u03bcM for measures of anti\u2010proliferative response, respectively. A relationship was observed between antiproliferative responses to HCQ and Lys05 and VPS34 CRISPR score with <jats:italic>D<\/jats:italic><jats:sub>m<\/jats:sub> values correlating with VPS34 response (<jats:italic>r<\/jats:italic>\u2009=\u20090.968 and 0.887) in a species independent manner. The results show that a subset of cOSA and hOSA cell lines are autophagy\u2010dependent and sensitive to HCQ at pharmacologically\u2010relevant exposures.<\/jats:p>","DOI":"10.1111\/vco.12937","type":"journal-article","created":{"date-parts":[[2023,9,19]],"date-time":"2023-09-19T06:54:59Z","timestamp":1695106499000},"page":"726-738","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Sensitivity of osteosarcoma cell lines to autophagy inhibition as determined by pharmacologic and genetic manipulation"],"prefix":"10.1111","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3063-1669","authenticated-orcid":false,"given":"Daniel L.","family":"Gustafson","sequence":"first","affiliation":[{"name":"School of Biomedical Engineering Colorado State University  Fort Collins Colorado USA"},{"name":"Department of Clinical Sciences Colorado State University  Fort Collins Colorado USA"},{"name":"Flint Animal Cancer 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