Intracellular cathepsin C levels determine sensitivity of cells to leucyl‐leucine methyl ester‐triggered apoptosis. (1st May 2020)

• Record Type: Journal Article
• Authors: Kavčič, Nežka; Butinar, Miha; Sobotič, Barbara; Hafner Česen, Maruša; Petelin, Ana; Bojić, Lea; Zavašnik Bergant, Tina; Bratovš, Andreja; Reinheckel, Thomas; Turk, Boris
• Is Part Of: FEBS journal. Volume 287:Number 23(2020)
• Page Start: 5148
• Page End: 5166
• Publication Date: 2020-05-01
• Subjects: apoptosis -- cancer -- cathepsin C -- leucyl‐leucine methyl ester -- lysosomal membrane permeabilization; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pathology, Molecular -- Periodicals; 572
• DOI: 10.1111/febs.15326
• Languages: English
• ISSNs: 1742-464X

Abstract

Abstract : L‐leucyl‐leucine methyl ester (LLOMe) is a lysosomotropic detergent, which was evaluated in clinical trials in graft‐vs‐host 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‐937 and THP‐1 promonocytes as models for monocytic cells, U‐87‐MG and HeLa cells as models for cancer cells, and noncancerous HEK293 cells, we show that the drug triggers rapid cathepsin C‐dependent 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—the most upstream molecule in the pathway—in 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. Abstract : The mechanism of action of LLOMe, a widely used lysosomotropic detergent, was studied in different cell models. We show that the rapid lysosomal membrane permeabilization is critically dependent on cathepsin C and that the levels of cathepsin C determine the sensitivity of cells to the detergent. Higher levels of other cathepsins are crucial for caspase activation and apoptosis, whereas high levels of cathepsin inhibitor stefin B protect against apoptosis.