TRPML3‑mediated lysosomal Ca<sup>2+</sup> release enhances drug sequestration and biogenesis, promoting osimertinib resistance in non‑small cell lung cancer

Kim,Mi Seong; Kim,Min Seuk

doi:10.3892/or.2025.8946

September-2025 Volume 54 Issue 3

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September-2025 Volume 54 Issue 3

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Article Open Access

TRPML3‑mediated lysosomal Ca²⁺ release enhances drug sequestration and biogenesis, promoting osimertinib resistance in non‑small cell lung cancer

Authors:
- Mi Seong Kim
- Min Seuk Kim
View Affiliations / Copyright

Affiliations: Department of Oral Physiology, Institute of Biomaterial‑Implant, School of Dentistry, Wonkwang University, Iksan, Jeonbuk 54538, Republic of Korea

Copyright: © Kim et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
Article Number: 113
|
Published online on: July 14, 2025

https://doi.org/10.3892/or.2025.8946
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Abstract

Lysosomes and lysosomal Ca²⁺ play crucial roles in cellular homeostasis and drug resistance. The lysosomal Ca²⁺ channel transient receptor potential mucolipin 3 (TRPML3; also known as mucolipin‑3 or MCOLN3) is a key regulator of autophagy and membrane trafficking; however, its role in tyrosine kinase inhibitor (TKI) resistance remains unclear. The contribution of TRPML3 to osimertinib resistance in non‑small cell lung cancer (NSCLC) was therefore assessed. Using publicly available RNA sequencing data, including profiles from clinical samples before and after osimertinib treatment, TRPML3 expression was measured in lung adenocarcinoma (LUAD) tissues. Additionally, two‑dimensional cell culture of, and three‑dimensional spheroids derived from, NSCLC cell lines were used to elucidate roles of TRPML3 in drug resistance. TRPML3 expression was significantly upregulated in both LUAD tissues from patients with residual disease after osimertinib treatment, as well as in osimertinib‑resistant NSCLC cells. TRPML3 knockdown in resistant PC9 cells restored sensitivity to osimertinib and multiple TKIs; this was replicated in spheroid models. Mechanistically, osimertinib induced intracellular Ca²⁺ oscillations in PC9 cells via lysosomal Ca²⁺ release through TRPML3 rather than through TRPML1. In summary, the present findings suggest that elevated TRPML3 expression compensates for TRPML1 to maintain lysosomal acidity and biogenesis during TKI treatment, facilitating drug sequestration and resistance and identifying TRPML3 as a potential target for overcoming osimertinib resistance in NSCLC.

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1	Zhang Z, Yue P, Lu T, Wang Y, Wei Y and Wei X: Role of lysosomes in physiological activities, diseases, and therapy. J Hematol Oncol. 14:792021. View Article : Google Scholar : PubMed/NCBI
2	Savini M, Zhao Q and Wang MC: Lysosomes: Signaling hubs for metabolic sensing and longevity. Trends Cell Biol. 29:876–887. 2019. View Article : Google Scholar : PubMed/NCBI
3	Hraběta J, Belhajová M, Šubrtová H, Merlos Rodrigo MA, Heger Z and Eckschlager T: Drug sequestration in lysosomes as one of the mechanisms of chemoresistance of cancer cells and the possibilities of its inhibition. Int J Mol Sci. 21:43922020. View Article : Google Scholar : PubMed/NCBI
4	Yamagishi T, Sahni S, Sharp DM, Arvind A, Jansson PJ and Richardson DR: P-glycoprotein mediates drug resistance via a novel mechanism involving lysosomal sequestration. J Biol Chem. 288:31761–31771. 2013. View Article : Google Scholar : PubMed/NCBI
5	Seebacher N, Lane DJ, Richardson DR and Jansson PJ: Turning the gun on cancer: Utilizing lysosomal P-glycoprotein as a new strategy to overcome multi-drug resistance. Free Radic Biol Med. 96:432–445. 2016. View Article : Google Scholar : PubMed/NCBI
6	Zhitomirsky B and Assaraf YG: Lysosomes as mediators of drug resistance in cancer. Drug Resist Updat. 24:23–33. 2016. View Article : Google Scholar : PubMed/NCBI
7	Xu H and Ren D: Lysosomal physiology. Annu Rev Physiol. 77:57–80. 2015. View Article : Google Scholar : PubMed/NCBI
8	Kendall RL and Holian A: The role of lysosomal ion channels in lysosome dysfunction. Inhal Toxicol. 33:41–54. 2021. View Article : Google Scholar : PubMed/NCBI
9	Xu M and Dong XP: Endolysosomal TRPMLs in cancer. Biomolecules. 11:652021. View Article : Google Scholar : PubMed/NCBI
10	Bargal R, Avidan N, Ben-Asher E, Olender Z, Zeigler M, Frumkin A, Raas-Rothschild A, Glusman G, Lancet D and Bach G: Identification of the gene causing mucolipidosis type IV. Nat Genet. 26:118–123. 2000. View Article : Google Scholar : PubMed/NCBI
11	Santoni G, Maggi F, Amantini C, Marinelli O, Nabissi M and Morelli MB: Pathophysiological role of transient receptor potential mucolipin channel 1 in calcium-mediated stress-induced neurodegenerative diseases. Front Physiol. 11:2512020. View Article : Google Scholar : PubMed/NCBI
12	Soyombo AA, Tjon-Kon-Sang S, Rbaibi Y, Bashllari E, Bisceglia J, Muallem S and Kiselyov K: TRP-ML1 regulates lysosomal pH and acidic lysosomal lipid hydrolytic activity. J Biol Chem. 281:7294–7301. 2006. View Article : Google Scholar : PubMed/NCBI
13	Di Paola S, Scotto-Rosato A and Medina DL: TRPML1: The Ca⁽²⁺⁾retaker of the lysosome. Cell Calcium. 69:112–121. 2018. View Article : Google Scholar : PubMed/NCBI
14	Qi J, Li Q, Xin T, Lu Q, Lin J, Zhang Y, Luo H, Zhang F, Xing Y, Wang W, et al: MCOLN1/TRPML1 in the lysosome: A promising target for autophagy modulation in diverse diseases. Autophagy. 20:1712–1722. 2024. View Article : Google Scholar : PubMed/NCBI
15	Medina DL, Di Paola S, Peluso I, Armani A, De Stefani D, Venditti R, Montefusco S, Scotto-Rosato A, Prezioso C, Forrester A, et al: Lysosomal calcium signalling regulates autophagy through calcineurin and TFEB. Nat Cell Biol. 17:288–299. 2015. View Article : Google Scholar : PubMed/NCBI
16	Kim SW, Kim DH, Park KS, Kim MK, Park YM, Muallem S, So I and Kim HJ: Palmitoylation controls trafficking of the intracellular Ca²⁺ channel MCOLN3/TRPML3 to regulate autophagy. Autophagy. 15:327–340. 2019. View Article : Google Scholar : PubMed/NCBI
17	Kim SW, Kim MK, Hong S, Choi A, Choi JH, Muallem S, So I, Yang D and Kim HJ: The intracellular Ca²⁺ channel TRPML3 is a PI3P effector that regulates autophagosome biogenesis. Proc Natl Acad Sci USA. 119:e22000851192022. View Article : Google Scholar : PubMed/NCBI
18	Paroutis P, Touret N and Grinstein S: The pH of the secretory pathway: Measurement, determinants, and regulation. Physiology (Bethesda). 19:207–215. 2004.PubMed/NCBI
19	Miao Y, Li G, Zhang X, Xu H and Abraham SN: A TRP channel senses lysosome neutralization by pathogens to trigger their expulsion. Cell. 161:1306–1319. 2015. View Article : Google Scholar : PubMed/NCBI
20	Gerasimenko JV, Tepikin AV, Petersen OH and Gerasimenko OV: Calcium uptake via endocytosis with rapid release from acidifying endosomes. Curr Biol. 8:1335–1338. 1998. View Article : Google Scholar : PubMed/NCBI
21	Martina JA, Lelouvier B and Puertollano R: The calcium channel mucolipin-3 is a novel regulator of trafficking along the endosomal pathway. Traffic. 10:1143–1156. 2009. View Article : Google Scholar : PubMed/NCBI
22	Liang J, Bi G, Sui Q, Zhao G, Zhang H, Bian Y, Chen Z, Huang Y, Xi J, Shi Y, et al: Transcription factor ZNF263 enhances EGFR-targeted therapeutic response and reduces residual disease in lung adenocarcinoma. Cell Rep. 43:1137712024. View Article : Google Scholar : PubMed/NCBI
23	Chen S, Zhou Y, Chen Y and Gu J: fastp: An ultra-fast all-in-one FASTQ preprocessor. Bioinformatics. 34:i884–i890. 2018. View Article : Google Scholar : PubMed/NCBI
24	Dobin A, Davis CA, Schlesinger F, Drenkow J, Zaleski C, Jha S, Batut P, Chaisson M and Gingeras TR: STAR: Ultrafast universal RNA-seq aligner. Bioinformatics. 29:15–21. 2013. View Article : Google Scholar : PubMed/NCBI
25	Patro R, Duggal G, Love MI, Irizarry RA and Kingsford C: Salmon provides fast and bias-aware quantification of transcript expression. Nat Methods. 14:417–419. 2017. View Article : Google Scholar : PubMed/NCBI
26	Robinson MD, McCarthy DJ and Smyth GK: edgeR: A Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics. 26:139–140. 2010. View Article : Google Scholar : PubMed/NCBI
27	Chu VT, Gottardo R, Raftery AE, Bumgarner RE and Yeung KY: MeV+R: Using MeV as a graphical user interface for Bioconductor applications in microarray analysis. Genome Biol. 9:R1182008. View Article : Google Scholar : PubMed/NCBI
28	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
29	Kurien BT and Scofield RH: Western blotting: An introduction. Methods Mol Biol. 1312:17–30. 2015. View Article : Google Scholar : PubMed/NCBI
30	Ramalingam SS, Vansteenkiste J, Planchard D, Cho BC, Gray JE, Ohe Y, Zhou C, Reungwetwattana T, Cheng Y, Chewaskulyong B, et al: Overall survival with osimertinib in untreated, EGFR-mutated advanced NSCLC. N Engl J Med. 382:41–50. 2020. View Article : Google Scholar : PubMed/NCBI
31	Ohashi K, Maruvka YE, Michor F and Pao W: Epidermal growth factor receptor tyrosine kinase inhibitor-resistant disease. J Clin Oncol. 31:1070–1080. 2013. View Article : Google Scholar : PubMed/NCBI
32	Halaby R: Influence of lysosomal sequestration on multidrug resistance in cancer cells. Cancer Drug Resist. 2:31–42. 2019.PubMed/NCBI
33	Zhitomirsky B and Assaraf YG: Lysosomal sequestration of hydrophobic weak base chemotherapeutics triggers lysosomal biogenesis and lysosome-dependent cancer multidrug resistance. Oncotarget. 6:1143–1156. 2015. View Article : Google Scholar : PubMed/NCBI
34	Wu M, Li X, Zhang T, Liu Z and Zhao Y: Identification of a nine-gene signature and establishment of a prognostic nomogram predicting overall survival of pancreatic cancer. Front Oncol. 9:9962019. View Article : Google Scholar : PubMed/NCBI
35	Kim HJ, Li Q, Tjon-Kon-Sang S, So I, Kiselyov K and Muallem S: Gain-of-function mutation in TRPML3 causes the mouse Varitint-Waddler phenotype. J Biol Chem. 282:36138–36142. 2007. View Article : Google Scholar : PubMed/NCBI
36	Kim HJ, Soyombo AA, Tjon-Kon-Sang S, So I and Muallem S: The Ca(2+) channel TRPML3 regulates membrane trafficking and autophagy. Traffic. 10:1157–1167. 2009. View Article : Google Scholar : PubMed/NCBI
37	Debnath J, Gammoh N and Ryan KM: Autophagy and autophagy-related pathways in cancer. Nat Rev Mol Cell Biol. 24:560–575. 2023. View Article : Google Scholar : PubMed/NCBI
38	Kim MS, Yang SH and Kim MS: TRPML3 enhances drug resistance in non-small cell lung cancer cells by promoting Ca²⁺-mediated lysosomal trafficking. Biochem Biophys Res Commun. 627:152–159. 2022. View Article : Google Scholar : PubMed/NCBI
39	Christensen KA, Myers JT and Swanson JA: pH-dependent regulation of lysosomal calcium in macrophages. J Cell Sci. 115:599–607. 2002. View Article : Google Scholar : PubMed/NCBI
40	Morgan AJ, Platt FM, Lloyd-Evans E and Galione A: Molecular mechanisms of endolysosomal Ca2+ signalling in health and disease. Biochem J. 439:349–374. 2011. View Article : Google Scholar : PubMed/NCBI
41	Shen D, Wang X, Li X, Zhang X, Yao Z, Dibble S, Dong XP, Yu T, Lieberman AP, Showalter HD and Xu H: Lipid storage disorders block lysosomal trafficking by inhibiting a TRP channel and lysosomal calcium release. Nat Commun. 3:7312012. View Article : Google Scholar : PubMed/NCBI
42	de Araujo MEG, Liebscher G, Hess MW and Huber LA: Lysosomal size matters. Traffic. 21:60–75. 2020. View Article : Google Scholar : PubMed/NCBI

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