{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,7]],"date-time":"2026-09-07T19:23:47Z","timestamp":1788809027964,"version":"build-2803163510"},"reference-count":63,"publisher":"Wiley","issue":"23","license":[{"start":{"date-parts":[[2020,5,1]],"date-time":"2020-05-01T00:00:00Z","timestamp":1588291200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["RE1584\/6\u20102"],"award-info":[{"award-number":["RE1584\/6\u20102"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["SFB 850 project B7"],"award-info":[{"award-number":["SFB 850 project B7"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100004329","name":"Javna Agencija za Raziskovalno Dejavnost RS","doi-asserted-by":"publisher","award":["P1\u20100140"],"award-info":[{"award-number":["P1\u20100140"]}],"id":[{"id":"10.13039\/501100004329","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The FEBS Journal"],"published-print":{"date-parts":[[2020,12]]},"abstract":"<jats:p>L\u2010leucyl\u2010leucine methyl ester (LLOMe) is a lysosomotropic detergent, which was evaluated in clinical trials in graft\u2010vs\u2010host disease because it very efficiently killed monocytic cell lines. It was also shown to efficiently trigger apoptosis in cancer cells, suggesting that the drug might have potential in anticancer therapy. Using U\u2010937 and THP\u20101 promonocytes as models for monocytic cells, U\u201087\u2010MG and HeLa cells as models for cancer cells, and noncancerous HEK293 cells, we show that the drug triggers rapid cathepsin C\u2010dependent lysosomal membrane permeabilization, followed by the release of other cysteine cathepsins into the cytosol and subsequent apoptosis. However, monocytes were found to be far more sensitive to the drug than the cancer and noncancer cells, which is most likely a consequence of the much higher intracellular levels of cathepsin C\u2014the most upstream molecule in the pathway\u2014in monocytic cell lines as compared to cancer cells. Overexpression of cathepsin C in HEK293 cells substantially enhances their sensitivity to the drug, consistent with the crucial role of cathepsin C. Major involvement of cysteine cathepsins B, S, and L in the downstream signaling pathway to mitochondrial cell death was confirmed in two gene ablation models, including the ablation of the major cytosolic inhibitor of cysteine cathepsins, stefin B, in primary mouse cancer cells, and simultaneous ablation of two major cathepsins, B and L, in mouse embryonic fibroblasts (MEFs). Deletion of stefin B resulted in sensitizing primary murine breast cancer cells to cell death without affecting the release of cathepsins, whereas simultaneous ablation of cathepsins B and L largely protected MEFs against cell death. However, due to the extreme sensitivity of monocytes to LLOMe, it appears that the drug may not be suitable for anticancer therapy due to risk of systemic toxicity.<\/jats:p>","DOI":"10.1111\/febs.15326","type":"journal-article","created":{"date-parts":[[2020,4,22]],"date-time":"2020-04-22T11:04:11Z","timestamp":1587553451000},"page":"5148-5166","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":37,"title":["Intracellular cathepsin C levels determine sensitivity of cells to leucyl\u2010leucine methyl ester\u2010triggered apoptosis"],"prefix":"10.1111","volume":"287","author":[{"given":"Ne\u017eka","family":"Kav\u010di\u010d","sequence":"first","affiliation":[{"name":"Department of Biochemistry, Molecular and Structural Biology Jo\u017eef Stefan Institute Ljubljana Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Miha","family":"Butinar","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Molecular and Structural Biology Jo\u017eef Stefan Institute Ljubljana Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Soboti\u010d","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Molecular and Structural Biology Jo\u017eef Stefan Institute Ljubljana Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maru\u0161a","family":"Hafner \u010cesen","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Molecular and Structural Biology Jo\u017eef Stefan Institute Ljubljana Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana","family":"Petelin","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Molecular and Structural Biology Jo\u017eef Stefan Institute Ljubljana Slovenia"},{"name":"Faculty of Health Sciences University of Primorska Izola Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lea","family":"Boji\u0107","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Molecular and Structural Biology Jo\u017eef Stefan Institute Ljubljana Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tina","family":"Zava\u0161nik Bergant","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Molecular and Structural Biology Jo\u017eef Stefan Institute Ljubljana Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreja","family":"Bratov\u0161","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Molecular and Structural Biology Jo\u017eef Stefan Institute Ljubljana Slovenia"},{"name":"Jo\u017eef Stefan International Postgraduate School Ljubljana Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Reinheckel","sequence":"additional","affiliation":[{"name":"Medical Faculty Institute of Molecular Medicine and Cell Research Albert\u2010Ludwigs University Freiburg Germany"},{"name":"German Cancer Consortium (DKTK) partner site Freiburg German Cancer Research Center (DKFZ) Heidelberg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9007-5764","authenticated-orcid":false,"given":"Boris","family":"Turk","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Molecular and Structural Biology Jo\u017eef Stefan Institute Ljubljana Slovenia"},{"name":"Faculty of Chemistry and Chemical Technology Ljubljana Slovenia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2020,5]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.1093\/emboj\/20.17.4629"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/emboj.2012.42"},{"key":"e_1_2_9_4_1","first-page":"141","article-title":"Structural and functional aspects of papain\u2010like cysteine proteinases and their protein inhibitors","volume":"378","author":"Turk B","year":"1997","journal-title":"Biol Chem"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.2741\/3089"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1515\/hsz-2016-0252"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1111\/tra.12613"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.97.6.1841"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.104.5.1223"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2010.12.043"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0014-5793(00)01286-2"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.82.8.2468"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.87.1.83"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1009695221038"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M308347200"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M802513200"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.148.12.3950"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.2000.2836"},{"key":"e_1_2_9_19_1","first-page":"678","article-title":"Ex vivo purging of allogeneic marrow with L\u2010Leucyl\u2010L\u2010leucine methyl ester","volume":"60","author":"Rosenfeld CS","year":"1995","journal-title":"A phase I study. Transplantation"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbmt.2009.08.020"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamcr.2008.09.008"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.R109.023820"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.yexcr.2012.03.005"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.091181"},{"key":"e_1_2_9_25_1","doi-asserted-by":"publisher","DOI":"10.1111\/nyas.12966"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mito.2014.06.006"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0076-6879(08)01409-2"},{"key":"e_1_2_9_28_1","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.204529"},{"key":"e_1_2_9_29_1","doi-asserted-by":"publisher","DOI":"10.2174\/1381612023394124"},{"key":"e_1_2_9_30_1","doi-asserted-by":"publisher","DOI":"10.1038\/onc.2013.314"},{"key":"e_1_2_9_31_1","doi-asserted-by":"publisher","DOI":"10.4161\/cc.24903"},{"key":"e_1_2_9_32_1","doi-asserted-by":"publisher","DOI":"10.1042\/BST20170130"},{"key":"e_1_2_9_33_1","doi-asserted-by":"publisher","DOI":"10.1097\/00007890-199206000-00030"},{"key":"e_1_2_9_34_1","doi-asserted-by":"publisher","DOI":"10.1038\/ni.1631"},{"key":"e_1_2_9_35_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M008944200"},{"key":"e_1_2_9_36_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M112.400655"},{"key":"e_1_2_9_37_1","doi-asserted-by":"publisher","DOI":"10.4161\/15384101.2014.991194"},{"key":"e_1_2_9_38_1","doi-asserted-by":"publisher","DOI":"10.1189\/jlb.0506359"},{"key":"e_1_2_9_39_1","doi-asserted-by":"publisher","DOI":"10.1038\/emboj.2013.171"},{"key":"e_1_2_9_40_1","doi-asserted-by":"publisher","DOI":"10.1101\/pdb.prot087114"},{"key":"e_1_2_9_41_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.aar5078"},{"key":"e_1_2_9_42_1","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-05-4463"},{"key":"e_1_2_9_43_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2011.02.013"},{"key":"e_1_2_9_44_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biochi.2007.10.004"},{"key":"e_1_2_9_45_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0907240107"},{"key":"e_1_2_9_46_1","doi-asserted-by":"publisher","DOI":"10.1038\/onc.2008.59"},{"key":"e_1_2_9_47_1","doi-asserted-by":"publisher","DOI":"10.1101\/gad.1407406"},{"key":"e_1_2_9_48_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrc4027"},{"key":"e_1_2_9_49_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1535-6108(04)00111-4"},{"key":"e_1_2_9_50_1","doi-asserted-by":"publisher","DOI":"10.1101\/gad.180331.111"},{"key":"e_1_2_9_51_1","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-07-0602"},{"key":"e_1_2_9_52_1","doi-asserted-by":"publisher","DOI":"10.1038\/nnano.2011.112"},{"key":"e_1_2_9_53_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biochi.2010.04.023"},{"key":"e_1_2_9_54_1","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-05-0269"},{"key":"e_1_2_9_55_1","doi-asserted-by":"publisher","DOI":"10.1038\/cddis.2013.361"},{"key":"e_1_2_9_56_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0002-9440(10)61253-9"},{"key":"e_1_2_9_57_1","doi-asserted-by":"publisher","DOI":"10.1002\/0471142727.mb2801s70"},{"key":"e_1_2_9_58_1","article-title":"Quantitation of apoptosis and necrosis by annexin V binding, propidium iodide uptake, and flow cytometry","volume":"2016","author":"Crowley LC","year":"2016","journal-title":"Cold Spring Harb Protoc"},{"key":"e_1_2_9_59_1","doi-asserted-by":"publisher","DOI":"10.1007\/s10495-007-0756-2"},{"key":"e_1_2_9_60_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-3581-9_5"},{"key":"e_1_2_9_61_1","doi-asserted-by":"publisher","DOI":"10.1101\/pdb.prot087106"},{"key":"e_1_2_9_62_1","doi-asserted-by":"publisher","DOI":"10.1515\/bchm3.1995.376.7.401"},{"key":"e_1_2_9_63_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biochi.2011.10.018"},{"key":"e_1_2_9_64_1","doi-asserted-by":"publisher","DOI":"10.1186\/1472-6750-8-91"}],"container-title":["The FEBS Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Ffebs.15326","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/febs.15326","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1111\/febs.15326","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/febs.15326","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,5]],"date-time":"2023-09-05T00:46:56Z","timestamp":1693874816000},"score":1,"resource":{"primary":{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/10.1111\/febs.15326"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,5]]},"references-count":63,"journal-issue":{"issue":"23","published-print":{"date-parts":[[2020,12]]}},"alternative-id":["10.1111\/febs.15326"],"URL":"https:\/\/doi.org\/10.1111\/febs.15326","archive":["Portico"],"relation":{},"ISSN":["1742-464X","1742-4658"],"issn-type":[{"value":"1742-464X","type":"print"},{"value":"1742-4658","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,5]]},"assertion":[{"value":"2019-07-04","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2020-03-05","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2020-05-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}