{"doi":"10.1073/pnas.2521979122","title":"NPC1 trafficking via VPS41-dependent LAMP carriers regulates endosomal cholesterol homeostasis","abstract":"The Niemann-Pick type 1 and 2 proteins (NPC1 and NPC2) coordinate cholesterol egress from late endosomes-lysosomes (LE/LY). Proper folding, trafficking, and localization of both NPC proteins are essential for normal LE/LY cholesterol handling. Accordingly, mutations in NPC genes cause Niemann–Pick type C (NPC) disease, a progressive neurodegenerative lysosomal cholesterol storage disorder. The routes by which NPC1 reaches the LE/LY compartment in mammalian cells are not fully elucidated. Therefore, to interrogate NPC1 trafficking, we developed genome-engineered HeLa cells expressing endogenous NPC1 mNeon . We demonstrate that endogenous NPC1 localizes to the LE/LY compartment and by using protein proximity-based approaches that NPC1 resides in the same membranes as Vacuolar Protein Sorting-associated protein 41 (VPS41), one of the two unique subunits of the homotypic fusion and vacuole protein sorting complex. Loss of VPS41 increases NPC1 and Lysosomal Associated Membrane Protein 1 (LAMP1) abundance. Paradoxically, this results in marked accumulation of lysosomal cholesterol and induction of sterol regulatory element-binding protein signaling. Mechanistically, using immuno-fluorescence and electron microscopy imaging in combination with a VPS41-dependent ectopic recruitment assay, we demonstrate that this is due to a shift in the localization of NPC1 and LAMP1 from LE/LY to biosynthetic vesicles called LAMP carriers. These vesicles have been recently described to transport lysosomal-destined cargo directly from the trans- Golgi (TGN) network to LE/LY. In conclusion, we identify NPC1 as a cargo for VPS41-dependent LAMP carriers that are instrumental for the delivery of NPC1 to LE/LY and maintaining cellular cholesterol homeostasis.","journal":"Proceedings of the National Academy of Sciences","year":2025,"id":535898,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":2,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9495,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1420656,"name":"Matteo Tantucci","orcid":null,"position":1,"is_corresponding":false},{"id":1420657,"name":"Paolo Sanza","orcid":null,"position":2,"is_corresponding":false},{"id":1420658,"name":"Kristian Zubak","orcid":null,"position":3,"is_corresponding":false},{"id":1420111,"name":"Giorgia Marodin","orcid":"0009-0008-2626-625X","position":4,"is_corresponding":false},{"id":1251937,"name":"Jenina Kingma","orcid":"0000-0003-2822-5415","position":5,"is_corresponding":false},{"id":1420659,"name":"Felix Snijder","orcid":null,"position":6,"is_corresponding":false},{"id":581002,"name":"Tineke Veenendaal","orcid":null,"position":7,"is_corresponding":false},{"id":484561,"name":"Daniel L. Kober","orcid":"0000-0001-8657-9348","position":8,"is_corresponding":false},{"id":257522,"name":"Noam Zelcer","orcid":"0000-0001-6935-7532","position":9,"is_corresponding":false},{"id":560410,"name":"Judith Klumperman","orcid":"0000-0003-4835-6228","position":10,"is_corresponding":false},{"id":1251936,"name":"Klevis Ndoj","orcid":"0009-0002-5902-7673","position":0,"is_corresponding":true}],"reference_count":46,"raw_metadata":null,"created_at":"2026-07-19T02:52:00.885532Z","pmid":"41452985","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}