Fusion of Lysosomes to Plasma Membrane Initiates Radiation-induced Apoptosis

Overview

Journal J Cell Biol

Specialty Cell Biology

Date 2020 Apr 25

PMID 32328634

Citations 20

Authors

Charles S Ferranti

Jin Cheng

Chris Thompson

Jianjun Zhang

Jimmy A Rotolo

Salma Buddaseth

Zvi Fuks

Richard N Kolesnick

Affiliations

Soon will be listed here.

Abstract

Diverse stresses, including reactive oxygen species (ROS), ionizing radiation, and chemotherapies, activate acid sphingomyelinase (ASMase) and generate the second messenger ceramide at plasma membranes, triggering apoptosis in specific cells, such as hematopoietic cells and endothelium. Ceramide elevation drives local bilayer reorganization into ceramide-rich platforms, macrodomains (0.5-5-µm diameter) that transmit apoptotic signals. An unresolved issue is how ASMase residing within lysosomes is released extracellularly within seconds to hydrolyze sphingomyelin preferentially enriched in outer plasma membranes. Here we show that physical damage by ionizing radiation and ROS induces full-thickness membrane disruption that allows local calcium influx, membrane lysosome fusion, and ASMase release. Further, electron microscopy reveals that plasma membrane "nanopore-like" structures (∼100-nm diameter) form rapidly due to lipid peroxidation, allowing calcium entry to initiate lysosome fusion. We posit that the extent of upstream damage to mammalian plasma membranes, calibrated by severity of nanopore-mediated local calcium influx for lysosome fusion, represents a biophysical mechanism for cell death induction.

Citing Articles

Emerging roles of the acid sphingomyelinase/ceramide pathway in metabolic and cardiovascular diseases: Mechanistic insights and therapeutic implications.

Wang H, Wang Y, Zhang S, Bai L World J Cardiol. 2025; 17(2):102308.

PMID: 40061281 PMC: 11886385. DOI: 10.4330/wjc.v17.i2.102308.

DNA2 knockout aggravates cerebral ischemia/reperfusion injury by reducing postsynaptic Homer1a.

Ma T, Li Y, Ren P, Wang S, Liu X, Lv W Zool Res. 2025; 46(1):87-102.

PMID: 39846189 PMC: 11890998. DOI: 10.24272/j.issn.2095-8137.2024.269.

Anti-Ceramide ScFv Prophylaxis for First Responders to a Limited Nuclear Attack.

Cheng J, Nagesh P, Feldman R, Shamu T, Zhang Z, Fuks Z Cell Physiol Biochem. 2024; 58(4):418-430.

PMID: 39172137 PMC: 11650686. DOI: 10.33594/000000721.

Combination of ionizing radiation and 2-thio-6-azauridine induces cell death in radioresistant triple negative breast cancer cells by downregulating CD151 expression.

Marni R, Malla M, Chakraborty A, Voonna M, Bhattacharyya P, Kgk D Cancer Chemother Pharmacol. 2024; 94(5):685-706.

PMID: 39167147 DOI: 10.1007/s00280-024-04709-w.

Key molecular DNA damage responses of human cells to radiation.

Zhang C, Liu J, Wu J, Ranjan K, Cui X, Wang X Front Cell Dev Biol. 2024; 12:1422520.

PMID: 39050891 PMC: 11266142. DOI: 10.3389/fcell.2024.1422520.

References

Stancevic B, Varda-Bloom N, Cheng J, Fuller J, Rotolo J, Garcia-Barros M . Adenoviral transduction of human acid sphingomyelinase into neo-angiogenic endothelium radiosensitizes tumor cure. PLoS One. 2013; 8(8):e69025. PMC: 3732255. DOI: 10.1371/journal.pone.0069025. View

Cremesti A, Paris F, Grassme H, Holler N, Tschopp J, Fuks Z . Ceramide enables fas to cap and kill. J Biol Chem. 2001; 276(26):23954-61. DOI: 10.1074/jbc.M101866200. View

Stancevic B, Kolesnick R . Ceramide-rich platforms in transmembrane signaling. FEBS Lett. 2010; 584(9):1728-40. PMC: 4440589. DOI: 10.1016/j.febslet.2010.02.026. View

Holopainen J, Angelova M, Kinnunen P . Vectorial budding of vesicles by asymmetrical enzymatic formation of ceramide in giant liposomes. Biophys J. 2000; 78(2):830-8. PMC: 1300685. DOI: 10.1016/S0006-3495(00)76640-9. View

Singh V, Beattie L, Seed T . Vitamin E: tocopherols and tocotrienols as potential radiation countermeasures. J Radiat Res. 2013; 54(6):973-88. PMC: 3823775. DOI: 10.1093/jrr/rrt048. View

Reddy A, Caler E, Andrews N . Plasma membrane repair is mediated by Ca(2+)-regulated exocytosis of lysosomes. Cell. 2001; 106(2):157-69. DOI: 10.1016/s0092-8674(01)00421-4. View

Li X, Gulbins E, Zhang Y . Oxidative stress triggers Ca-dependent lysosome trafficking and activation of acid sphingomyelinase. Cell Physiol Biochem. 2012; 30(4):815-26. PMC: 3777434. DOI: 10.1159/000341460. View

Radford I, Hodgson G . 125I-induced DNA double strand breaks: use in calibration of the neutral filter elution technique and comparison with X-ray induced breaks. Int J Radiat Biol Relat Stud Phys Chem Med. 1985; 48(4):555-66. DOI: 10.1080/09553008514551611. View

van der Goot F, Harder T . Raft membrane domains: from a liquid-ordered membrane phase to a site of pathogen attack. Semin Immunol. 2001; 13(2):89-97. DOI: 10.1006/smim.2000.0300. View

10.

Nurminen T, Holopainen J, Zhao H, Kinnunen P . Observation of topical catalysis by sphingomyelinase coupled to microspheres. J Am Chem Soc. 2002; 124(41):12129-34. DOI: 10.1021/ja017807r. View

11.

Grassme H, Jendrossek V, Bock J, Riehle A, Gulbins E . Ceramide-rich membrane rafts mediate CD40 clustering. J Immunol. 2001; 168(1):298-307. DOI: 10.4049/jimmunol.168.1.298. View

12.

Narayan K, Subramaniam S . Focused ion beams in biology. Nat Methods. 2015; 12(11):1021-31. PMC: 6993138. DOI: 10.1038/nmeth.3623. View

13.

Powell S, McMILLAN T . The repair fidelity of restriction enzyme-induced double strand breaks in plasmid DNA correlates with radioresistance in human tumor cell lines. Int J Radiat Oncol Biol Phys. 1994; 29(5):1035-40. DOI: 10.1016/0360-3016(94)90399-9. View

14.

Jeggo P, Lobrich M . How cancer cells hijack DNA double-strand break repair pathways to gain genomic instability. Biochem J. 2015; 471(1):1-11. DOI: 10.1042/BJ20150582. View

15.

Rotolo J, Zhang J, Donepudi M, Lee H, Fuks Z, Kolesnick R . Caspase-dependent and -independent activation of acid sphingomyelinase signaling. J Biol Chem. 2005; 280(28):26425-34. DOI: 10.1074/jbc.M414569200. View

16.

Belka C, Gruber C, Jendrossek V, Wesselborg S, Budach W . The tyrosine kinase Lck is involved in regulation of mitochondrial apoptosis pathways. Oncogene. 2003; 22(2):176-85. DOI: 10.1038/sj.onc.1206103. View

17.

Defour A, van der Meulen J, Bhat R, Bigot A, Bashir R, Nagaraju K . Dysferlin regulates cell membrane repair by facilitating injury-triggered acid sphingomyelinase secretion. Cell Death Dis. 2014; 5:e1306. PMC: 4079937. DOI: 10.1038/cddis.2014.272. View

18.

LeMotte P, Little J . DNA damage induced in human diploid cells by decay of incorporated radionuclides. Cancer Res. 1984; 44(4):1337-42. View

19.

Stark G . The effect of ionizing radiation on lipid membranes. Biochim Biophys Acta. 1991; 1071(2):103-22. DOI: 10.1016/0304-4157(91)90020-w. View

20.

Dutta D, Williamson C, Cole N, Donaldson J . Pitstop 2 is a potent inhibitor of clathrin-independent endocytosis. PLoS One. 2012; 7(9):e45799. PMC: 3448704. DOI: 10.1371/journal.pone.0045799. View