{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,21]],"date-time":"2026-04-21T06:38:44Z","timestamp":1776753524303,"version":"3.51.2"},"reference-count":45,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2026,4,20]],"date-time":"2026-04-20T00:00:00Z","timestamp":1776643200000},"content-version":"vor","delay-in-days":109,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T00:00:00Z","timestamp":1767225600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100001691","name":"Japan Society for the Promotion of Science","doi-asserted-by":"publisher","award":["23K07437"],"award-info":[{"award-number":["23K07437"]}],"id":[{"id":"10.13039\/501100001691","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Immunology Research"],"published-print":{"date-parts":[[2026,1]]},"abstract":"<jats:p>\n                    The tumor microenvironment (TME) provides a niche for immune evasion and immunotherapy resistance, in part, by recruiting pro\u2010tumor M2\u2010like macrophages. In the present study, using heterospheroids consisting of cancer cells and macrophages, we identified TAM activators, which are compounds that reprogram M2\u2010like tumor\u2010associated macrophages (TAMs) toward the antitumor M1\u2010like phenotype. THP\u20101\u2010 or human peripheral monocyte\u2010derived macrophages were co\u2010cultured with liver cancer (LC) cells in an ultra\u2010low attachment dish to generate heterospheroids. Cell surface marker expression and macrophage infiltration into the heterospheroids were assessed by flow cytometry and fluorescence microscopy, respectively. Lipopolysaccharide (LPS) and interferon\u2010\u03b3 (IFN\u03b3)\u2010induced M1 marker expression was observed on the macrophages in the homospheroids; however, this induction was suppressed in heterospheroids. Microscopic imaging revealed that macrophage infiltration into the heterospheroids was decreased in the presence of LPS and IFN\u03b3, which prompted us to develop a high\u2010content imaging screen. We identified two compounds [alprostadil (prostaglandin E1) and HX531] with TAM\u2010activating activity. RNA\u2010seq analysis revealed that HX531 modulated the immune and IFN response in cancer cells and cell division in macrophages. Moreover, alprostadil promoted the M1\u2010like polarization of TAMs, increased tumor\u2010infiltrating CD8\n                    <jats:sup>+<\/jats:sup>\n                    T cells, and enhanced anti\u2010PD\u20101 antibody therapeutic efficacy in a syngeneic tumor\u2010bearing mouse. In conclusion, the heterospheroid culture recapitulates the immunosuppressive TME, which prevents the M1 polarization of TAMs. It provides a new platform for screening TAM activators and will enable the development of novel cancer immunotherapeutics when combined with high\u2010content imaging analysis.\n                  <\/jats:p>","DOI":"10.1155\/jimr\/2889509","type":"journal-article","created":{"date-parts":[[2026,4,21]],"date-time":"2026-04-21T05:41:31Z","timestamp":1776750091000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["High\u2010Throughput Heterospheroid\u2010Based Screening Identifies Drugs That Reprogram Tumor\u2010Associated 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