{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T10:14:38Z","timestamp":1785406478820,"version":"3.56.0"},"posted":{"date-parts":[[2025,11,13]]},"group-title":"Neuroscience","reference-count":26,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,11,13]],"date-time":"2025-11-13T00:00:00Z","timestamp":1762992000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"}],"funder":[{"award":["R01AG070202"],"award-info":[{"award-number":["R01AG070202"]}],"id":[{"id":"https:\/\/ror.org\/049v75w11","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,11,13]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Individuals who possess the \u03b54 allele of apolipoprotein E (\n                  <jats:italic>APOE<\/jats:italic>\n                  ) are at a significantly increased risk for developing Alzheimer\u2019s disease (AD). However, the precise reason for this association is not fully understood. Beyond its effects on amyloid and tau,\n                  <jats:italic>APOE<\/jats:italic>\n                  also influences fundamental cellular processes in the brain, including cholesterol trafficking between cells and within the endolysosomal system, which may be a critical component of the\n                  <jats:italic>APOE4<\/jats:italic>\n                  -associated vulnerability to AD. Here, we examined how\n                  <jats:italic>APOE<\/jats:italic>\n                  genotype, sex, and aging alter intracellular cholesterol processing and the endolysosomal system in the mouse brain. Using the novel cholesterol-binding probe D4H*-mCherry, we quantified intracellular cholesterol levels, the levels of early endosomes (Rab5), late endosomes (CD63), and lysosomes (LAMP1), and the colocalization of cholesterol with these endolysosomal compartments. This analysis was performed in the cortex, hippocampus, and entorhinal cortex of young, middle-aged, and old\n                  <jats:italic>APOE2<\/jats:italic>\n                  ,\n                  <jats:italic>APOE3<\/jats:italic>\n                  , and\n                  <jats:italic>APOE4<\/jats:italic>\n                  mice. Our analysis revealed region-specific changes in intracellular cholesterol and the endolysosomal system in response to aging, sex, and\n                  <jats:italic>APOE<\/jats:italic>\n                  genotype. Notably, young\n                  <jats:italic>APOE4<\/jats:italic>\n                  mice showed reduced cholesterol within early and late endosomes, but increased lysosomal abundance, suggesting impaired cholesterol processing. These APOE4-specific effects were less apparent in older animals. These effects were strongly modified by sex, with female\n                  <jats:italic>APOE4<\/jats:italic>\n                  mice exhibiting elevated lysosomal cholesterol in the hippocampus and entorhinal cortex at old age, indicating sex-dependent susceptibility. Together, these results reveal that\n                  <jats:italic>APOE<\/jats:italic>\n                  genotype, age, and sex interact to influence endolysosomal cholesterol homeostasis, with female\n                  <jats:italic>APOE4<\/jats:italic>\n                  mice showing the greatest dysregulation. These findings suggest that early and region-specific endolysosomal defects may contribute to the heightened AD risk associated with\n                  <jats:italic>APOE4<\/jats:italic>\n                  , particularly in females.\n                <\/jats:p>","DOI":"10.1101\/2025.11.11.687891","type":"posted-content","created":{"date-parts":[[2025,11,13]],"date-time":"2025-11-13T18:45:19Z","timestamp":1763059519000},"source":"Crossref","is-referenced-by-count":0,"title":["Investigating the Effects of\n                  <i>APOE<\/i>\n                  Genotype on Intracellular Cholesterol and the Endolysosomal System in the Aging Mouse Brain"],"prefix":"10.64898","author":[{"given":"Sherida M.","family":"de Leeuw","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tal","family":"Nuriel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2025111414101896000_2025.11.11.687891v1.1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.genom.1.1.507"},{"issue":"15","key":"2025111414101896000_2025.11.11.687891v1.2","doi-asserted-by":"crossref","first-page":"1896","DOI":"10.1007\/s00018-004-4009-z","article-title":"The role of apolipoprotein E in lipid metabolism in the central nervous system. Cell Mol Life Sci [Internet]","volume":"61","year":"2004","journal-title":"Cell Mol Life Sci"},{"key":"2025111414101896000_2025.11.11.687891v1.3","unstructured":"Holtzman DM , Herz J , Bu G. Apolipoprotein E and Apolipoprotein E Receptors: Normal Biology and Roles in Alzheimer Disease. [cited 2025 Nov 10]; Available from: http:\/\/perspectivesinmedicine.cshlp.org\/Downloadedfrom"},{"key":"2025111414101896000_2025.11.11.687891v1.4","doi-asserted-by":"publisher","DOI":"10.1186\/s13024-015-0001-3"},{"key":"2025111414101896000_2025.11.11.687891v1.5","doi-asserted-by":"publisher","DOI":"10.1093\/jnen\/nlw067"},{"issue":"18","key":"2025111414101896000_2025.11.11.687891v1.6","first-page":"10771","article-title":"Opposing effects of apolipoproteins E and C on lipoprotein binding to low density lipoprotein receptor-related protein","volume":"265","year":"1990","journal-title":"Journal of Biological Chemistry [Internet]. American Society for Biochemistry and Molecular Biology Inc"},{"key":"2025111414101896000_2025.11.11.687891v1.7","doi-asserted-by":"publisher","DOI":"10.1172\/JCI110330"},{"issue":"1","key":"2025111414101896000_2025.11.11.687891v1.8","doi-asserted-by":"crossref","first-page":"110","DOI":"10.1016\/j.stemcr.2021.11.007","article-title":"APOE2, E3, and E4 differentially modulate cellular homeostasis, cholesterol metabolism, and inflammatory response in isogenic iPSC-derived astrocytes. Stem Cell Reports [Internet]","volume":"17","year":"2022","journal-title":"Stem Cell Reports"},{"issue":"13","key":"2025111414101896000_2025.11.11.687891v1.9","first-page":"2213","article-title":"Cholesterol and matrisome pathways dysregulated in astrocytes and microglia","volume":"185","year":"2022","journal-title":"Cell. Cell Press"},{"key":"2025111414101896000_2025.11.11.687891v1.10","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M409324200"},{"key":"2025111414101896000_2025.11.11.687891v1.11","doi-asserted-by":"publisher","DOI":"10.1006\/nbdi.2000.0362"},{"key":"2025111414101896000_2025.11.11.687891v1.12","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M203934200"},{"key":"2025111414101896000_2025.11.11.687891v1.13","doi-asserted-by":"publisher","DOI":"10.3389\/fnins.2017.00702"},{"key":"2025111414101896000_2025.11.11.687891v1.14","article-title":"Cholesterol metabolism in aging simultaneously altered in liver and nervous system","volume":"14","year":"2022","journal-title":"Aging [Internet]"},{"key":"2025111414101896000_2025.11.11.687891v1.15","doi-asserted-by":"publisher","DOI":"10.1038\/s41380-018-0200-8"},{"key":"2025111414101896000_2025.11.11.687891v1.16","doi-asserted-by":"publisher","DOI":"10.1016\/j.molmed.2012.12.007"},{"key":"2025111414101896000_2025.11.11.687891v1.17","doi-asserted-by":"publisher","DOI":"10.1001\/jamaneurol.2023.3599"},{"key":"2025111414101896000_2025.11.11.687891v1.18","doi-asserted-by":"crossref","unstructured":"Maioli S , Nalvarte I , Ankarcrona M , Schultzberg M , Zuloaga KL , Goikolea J , Visser PJ , Strooper B De , Winblad B , Pizzo P , Williams PA , Rodriguez-Rodriguez P , Naia L . Bioenergetics and lipid metabolism in Alzheimer\u2019s disease: From cell biology to systemic health. J Intern Med [Internet]. John Wiley & Sons, Ltd; 2025 Nov 6 [cited 2025 Nov 10]; Available from: \/doi\/pdf\/10.1111\/joim.70036","DOI":"10.1111\/joim.70036"},{"key":"2025111414101896000_2025.11.11.687891v1.19","doi-asserted-by":"crossref","unstructured":"de Leeuw SM , Nuriel T. Intracellular cholesterol visualization in brain tissue using D4H\u2217. STAR Protoc. Cell Press; 2024 Mar 15;5(1). 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Available from: \/pmc\/articles\/PMC9219958\/","DOI":"10.3390\/biomedicines10061387"},{"key":"2025111414101896000_2025.11.11.687891v1.25","doi-asserted-by":"publisher","DOI":"10.1093\/cercor\/bhy109"},{"key":"2025111414101896000_2025.11.11.687891v1.26","doi-asserted-by":"publisher","DOI":"10.1038\/s43587-024-00769-y"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.1101\/2025.11.11.687891","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T10:01:01Z","timestamp":1768471261000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.1101\/2025.11.11.687891"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,13]]},"references-count":26,"URL":"https:\/\/doi.org\/10.1101\/2025.11.11.687891","relation":{},"subject":[],"published":{"date-parts":[[2025,11,13]]},"subtype":"preprint"}}