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Cancer cells in acidic environments increase lipid droplet accumulation and oxidative phosphorylation, which are characteristics of aggressive cancers. Here, we used live imaging, shotgun lipidomics and immunofluorescence analyses of mammary and pancreatic cancer cells to demonstrate that both acute acidosis and adaptation to acidic growth drive rapid uptake of fatty acids (FAs), which are converted to triacylglycerols and stored in lipid droplets. Consistent with being independent of de novo synthesis, triacylglycerol and lipid droplet accumulation in acid-adapted cells was unaffected by FA synthetase (FAS, encoded by FASN) inhibitors. Macropinocytosis, which is upregulated in acid-adapted cells, partially contributed to FA uptake, which was independent of other protein-facilitated lipid uptake mechanisms, including uptake via CD36 and FATP2, and caveolin- and clathrin-dependent endocytosis. We propose that a major mechanism by which tumor acidosis drives FA uptake is through neutralizing protonation of negatively charged FAs allowing their diffusive, transporter-independent uptake. We suggest that this could be a major factor triggering acidosis-driven metabolic rewiring.<\/jats:p>","DOI":"10.1242\/jcs.263688","type":"journal-article","created":{"date-parts":[[2025,4,7]],"date-time":"2025-04-07T07:42:01Z","timestamp":1744011721000},"source":"Crossref","is-referenced-by-count":7,"title":["Tumor acidosis supports cancer cell lipid uptake via a rapid transporter-independent mechanism"],"prefix":"10.1242","volume":"138","author":[{"given":"Marc","family":"Severin","sequence":"first","affiliation":[{"name":"University of Copenhagen 1 Section for Cell Biology and Physiology, Department of Biology , , 2100 Copenhagen ,","place":["Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rikke K.","family":"Hansen","sequence":"additional","affiliation":[{"name":"University of Copenhagen 1 Section for Cell Biology and Physiology, Department of Biology , , 2100 Copenhagen ,","place":["Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michala G.","family":"Rolver","sequence":"additional","affiliation":[{"name":"University of Copenhagen 1 Section for Cell Biology and Physiology, Department of Biology , , 2100 Copenhagen ,","place":["Denmark"]},{"name":"Biotech Research and Innovation Centre, University of Copenhagen 2 , 2200 Copenhagen ,","place":["Denmark"]},{"name":"University of Copenhagen 3 The Bioinformatics Centre, Department of Biology , , 2200 Copenhagen ,","place":["Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tove","family":"Hels","sequence":"additional","affiliation":[{"name":"University of Copenhagen 1 Section for Cell Biology and Physiology, Department of Biology , , 2100 Copenhagen ,","place":["Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kenji","family":"Maeda","sequence":"additional","affiliation":[{"name":"Danish Cancer Institute 4 Section for Cell Death and Metabolism , , 2100 Copenhagen ,","place":["Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Luis A.","family":"Pardo","sequence":"additional","affiliation":[{"name":"Max Planck Institute for Multidisciplinary Sciences 5 , Oncophysiology Group , 37077 G\u00f6ttingen ,","place":["Germany"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3044-7714","authenticated-orcid":false,"given":"Stine F.","family":"Pedersen","sequence":"additional","affiliation":[{"name":"University of Copenhagen 1 Section for Cell Biology and Physiology, Department of Biology , , 2100 Copenhagen ,","place":["Denmark"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"237","published-online":{"date-parts":[[2025,5,15]]},"reference":[{"key":"2025051604555655900_JCS263688C1","doi-asserted-by":"publisher","first-page":"586","DOI":"10.1002\/2211-5463.13559","article-title":"The breast cancer microenvironment and lipoprotein lipase: Another negative notch for a beneficial enzyme?","volume":"13","author":"Bavis","year":"2023","journal-title":"FEBS Open Biol."},{"key":"2025051604555655900_JCS263688C2","doi-asserted-by":"publisher","first-page":"108","DOI":"10.1038\/nature13110","article-title":"Metabolic determinants of cancer cell sensitivity to glucose limitation and biguanides","volume":"508","author":"Birsoy","year":"2014","journal-title":"Nature"},{"key":"2025051604555655900_JCS263688C3","doi-asserted-by":"publisher","first-page":"612","DOI":"10.1039\/C6MD00043F","article-title":"Fatty acid transport proteins: targeting FATP2 as a gatekeeper involved in the transport of exogenous fatty acids","volume":"7","author":"Black","year":"2016","journal-title":"Medchemcomm"},{"key":"2025051604555655900_JCS263688C63","doi-asserted-by":"publisher","first-page":"708","DOI":"10.3390\/ijms18040708","article-title":"\u201cBligh and Dyer\u201d and Folch methods for solid-liquid-liquid extraction of lipids from microorganisms. 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