Piezo1 channel exaggerates ferroptosis of nucleus pulposus cells by mediating mechanical stress-induced iron influx

Abstract

To date, several molecules have been found to facilitate iron influx, while the types of iron influx channels remain to be elucidated. Here, Piezo1 channel was identified as a key iron transporter in response to mechanical stress. Piezo1-mediated iron overload disturbed iron metabolism and exaggerated ferroptosis in nucleus pulposus cells (NPCs). Importantly, Piezo1-induced iron influx was independent of the transferrin receptor (TFRC), a well-recognized iron gatekeeper. Furthermore, pharmacological inactivation of Piezo1 profoundly reduced iron accumulation, alleviated mitochondrial ROS, and suppressed ferroptotic alterations in stimulation of mechanical stress. Moreover, conditional knockout of Piezo1 (Col2a1-CreERT Piezo1 ^flox/flox) attenuated the mechanical injury-induced intervertebral disc degeneration (IVDD). Notably, the protective effect of Piezo1 deficiency in IVDD was dampened in Piezo1/Gpx4 conditional double knockout (cDKO) mice (Col2a1-CreERT Piezo1 ^flox/flox /Gpx4 ^flox/flox). These findings suggest that Piezo1 is a potential determinant of iron influx, indicating that the Piezo1-iron-ferroptosis axis might shed light on the treatment of mechanical stress-induced diseases.

References

[1] Koleini N, Shapiro JS, Geier J, Ardehali H. Ironing out mechanisms of iron homeostasis and disorders of iron deficiency. J. Clin. Invest., 2021, 131: e148671

[2] Delijewski M, Bartoń A, Maksym B, Pawlas N. The link between iron turnover and pharmacotherapy in transplant patients. Nutrients., 2023, 15: 1453

[3] Sevimli G et al. Nitric oxide biosensor uncovers diminished ferrous iron-dependency of cultured cells adapted to physiological oxygen levels. Redox Biol., 2022, 53: 102319

[4] Donovan A et al. The iron exporter ferroportin/Slc40a1 is essential for iron homeostasis. Cell Metab., 2005, 1: 191-200

[5] Hassannia B, Vandenabeele P, Vanden Berghe T. Targeting ferroptosis to iron out cancer. Cancer Cell., 2019, 35: 830-849

[6] Vogt AS et al. On iron metabolism and its regulation. Int. J. Mol. Sci., 2021, 22: 4591

[7] Dixon SJ et al. Ferroptosis: an iron-dependent form of nonapoptotic cell death. Cell., 2012, 149: 1060-1072

[8] Yang WS, Stockwell BR. Ferroptosis: death by lipid peroxidation. Trends Cell Biol., 2016, 26: 165-176

[9] Wolfenson H, Yang B, Sheetz MP. Steps in mechanotransduction pathways that control cell morphology. Annu. Rev. Physiol., 2019, 81: 585-605

[10] Nazet U et al. Mechanical stress induce PG-E2 in murine synovial fibroblasts originating from the temporomandibular joint. Cells., 2021, 10: 298

[11] Onwudiwe K, Najera J, Siri S, Datta M. Do tumor mechanical stresses promote cancer immune escape? Cells., 2022, 11: 3840

[12] Höhfeld J et al. Maintaining proteostasis under mechanical stress. EMBO Rep., 2021, 22: e52507

[13] Shi S et al. Excessive mechanical stress-induced intervertebral disc degeneration is related to Piezo1 overexpression triggering the imbalance of autophagy/apoptosis in human nucleus pulpous. Arthritis Res. Ther., 2022, 24: 119

[14] Sonam S et al. Mechanical stress driven by rigidity sensing governs epithelial stability. Nat. Phys., 2023, 19: 132-141

[15] Lyons JS et al. Microtubules tune mechanotransduction through NOX2 and TRPV4 to decrease sclerostin abundance in osteocytes. Sci. Signal., 2017, 10: eaan5748

[16] Chen X, Xu S, Zhao C, Liu B. Role of TLR4/NADPH oxidase 4 pathway in promoting cell death through autophagy and ferroptosis during heart failure. Biochem. Biophys. Res. Commun., 2019, 516: 37-43

[17] Gottlieb PA, Sachs F. Piezo1: properties of a cation selective mechanical channel. Channels, 2012, 6: 214-219

[18] Niu L et al. A positive mechanobiological feedback loop controls bistable switching of cardiac fibroblast phenotype. Cell Discov., 2022, 8: 84

[19] Coste B et al. Piezo1 and Piezo2 are essential components of distinct mechanically activated cation channels. Science., 2010, 330: 55-60

[20] Solis AG et al. Mechanosensation of cyclical force by PIEZO1 is essential for innate immunity. Nature., 2019, 573: 69-74

[21] Sun W et al. The mechanosensitive Piezo1 channel is required for bone formation. Elife., 2019, 8: e47454

[22] Zhao X et al. Mechanosensitive Piezo1 channels mediate renal fibrosis. JCI Insight., 2022, 7: e152330

[23] Allison SJ. Hypertension: mechanosensation by PIEZO1 in blood pressure control. Nat. Rev. Nephrol., 2017, 13: 3

[24] Yang Q et al. Study on the mechanism of excessive apoptosis of nucleus pulposus cells induced by shRNA-Piezo1 under abnormal mechanical stretch stress. J. Cell Biochem., 2019, 120: 3989-3997

[25] Choi H et al. Hypoxia promotes noncanonical autophagy in nucleus pulposus cells independent of MTOR and HIF1A signaling. Autophagy, 2016, 12: 1631-1646

[26] Wang Y et al. SIRT1 alleviates high-magnitude compression-induced senescence in nucleus pulposus cells via PINK1-dependent mitophagy. Aging, 2020, 12: 16126-16141

[27] Vergroesen PP et al. Mechanics and biology in intervertebral disc degeneration: a vicious circle. Osteoarthr. Cartil., 2015, 23: 1057-1070

[28] Farfan HF. The torsional injury of the lumbar spine. Spine, 1984, 9: 53

[29] Iatridis JC, Setton LA, Weidenbaum M, Mow VC. Alterations in the mechanical behavior of the human lumbar nucleus pulposus with degeneration and aging. J. Orthop. Res., 1997, 15: 318-322

[30] Wu J et al. Self-amplifying loop of NF-κB and periostin initiated by PIEZO1 accelerates mechano-induced senescence of nucleus pulposus cells and intervertebral disc degeneration. Mol. Ther., 2022, 30: 3241-3256

[31] Wang S et al. Mechanical overloading induces GPX4-regulated chondrocyte ferroptosis in osteoarthritis via Piezo1 channel facilitated calcium influx. J. Adv. Res., 2022, 41: 63-75

[32] Dutt S, Hamza I, Bartnikas TB. Molecular mechanisms of iron and heme metabolism. Annu. Rev. Nutr., 2022, 42: 311-335

[33] Hirata Y et al. Lipid peroxidation increases membrane tension, Piezo1 gating, and cation permeability to execute ferroptosis. Curr. Biol., 2023, 33: 1282-1294.e5

[34] Stockwell BR et al. Ferroptosis: a regulated cell death nexus linking metabolism, redox biology, and disease. Cell., 2017, 171: 273-285

[35] Qiu S et al. Mitochondria-localized cGAS suppresses ferroptosis to promote cancer progression. Cell Res., 2023, 33: 299-311

[36] Yu F et al. Dynamic O-GlcNAcylation coordinates ferritinophagy and mitophagy to activate ferroptosis. Cell Discov., 2022, 8: 40

[37] Ma S et al. A role of PIEZO1 in iron metabolism in mice and humans. Cell., 2021, 184: 969-982.e13

[38] Gnanasambandam R, Bae C, Gottlieb PA, Sachs F. Ionic selectivity and permeation properties of human PIEZO1 channels. PLoS One, 2015, 10: e0125503

[39] Jin P et al. Electron cryo-microscopy structure of the mechanotransduction channel NOMPC. Nature., 2017, 547: 118-122

[40] Ge J et al. Architecture of the mammalian mechanosensitive Piezo1 channel. Nature., 2015, 527: 64-69

[41] Zhao Q et al. Structure and mechanogating mechanism of the Piezo1 channel. Nature., 2018, 554: 487-492

[42] Douguet D, Honoré E. Mammalian mechanoelectrical transduction: structure and function of force-gated ion channels. Cell., 2019, 179: 340-354

[43] Colombier P et al. The lumbar intervertebral disc: from embryonic development to degeneration. Joint Bone Spine., 2014, 81: 125-129

[44] Hall JA et al. Systematic review of decision analytic modelling in economic evaluations of low back pain and sciatica. Appl. Health Econ. Health Policy, 2019, 17: 467-491

[45] Zhu D et al. A new hope in spinal degenerative diseases: Piezo1. Biomed. Res. Int., 2021, 2021: 6645193

[46] Sun Y et al. Piezo1 activates the NLRP3 inflammasome in nucleus pulposus cell-mediated by Ca²⁺/NF-κB pathway. Int. Immunopharmacol., 2020, 85: 106681

[47] Doll S et al. ACSL4 dictates ferroptosis sensitivity by shaping cellular lipid composition. Nat. Chem. Biol., 2017, 13: 91-98

[48] Yan HF et al. Ferroptosis: mechanisms and links with diseases. Signal Transduct. Target Ther., 2021, 6: 49

[49] Alvarez SW et al. NFS1 undergoes positive selection in lung tumours and protects cells from ferroptosis. Nature., 2017, 551: 639-643

[50] Linkermann A et al. Synchronized renal tubular cell death involves ferroptosis. Proc. Natl. Acad. Sci. USA., 2014, 111: 16836-16841

[51] Wang H et al. Characterization of ferroptosis in murine models of hemochromatosis. Hepatology., 2017, 66: 449-465

[52] Wang W et al. Iron overload promotes intervertebral disc degeneration via inducing oxidative stress and ferroptosis in endplate chondrocytes. Free Radic Biol. Med., 2022, 190: 234-246

[53] Luck AN, Mason AB. Transferrin-mediated cellular iron delivery. Curr. Top. Membr., 2012, 69: 3-35

[54] Yu Y et al. Hepatic transferrin plays a role in systemic iron homeostasis and liver ferroptosis. Blood., 2020, 136: 726-739

[55] Gao M et al. Glutaminolysis and transferrin regulate ferroptosis. Mol. Cell., 2015, 59: 298-308

[56] Feng H et al. Transferrin receptor is a specific ferroptosis marker. Cell Rep., 2020, 30: 3411-3423.e7

[57] Pan J et al. The imbalance of p53-Park7 signaling axis induces iron homeostasis dysfunction in doxorubicin-challenged cardiomyocytes. Adv. Sci., 2023, 10: e2206007

[58] Zilka O et al. On the mechanism of cytoprotection by ferrostatin-1 and liproxstatin-1 and the role of lipid peroxidation in ferroptotic cell death. ACS Cent. Sci., 2017, 3: 232-243

[59] Doll S et al. FSP1 is a glutathione-independent ferroptosis suppressor. Nature., 2019, 575: 693-698

[60] Kannan M et al. HIV-1 Tat-mediated microglial ferroptosis involves the miR-204-ACSL4 signaling axis. Redox Biol., 2023, 62: 102689

[61] Koppula P et al. A targetable CoQ-FSP1 axis drives ferroptosis- and radiation-resistance in KEAP1 inactive lung cancers. Nat. Commun., 2022, 13

[62] Gottlieb PA. A tour de force: the discovery, properties, and function of piezo channels. Curr. Top Membr., 2017, 79: 1-36

[63] Martinac B, Poole K. Mechanically activated ion channels. Int. J. Biochem. Cell Biol., 2018, 97: 104-107

[64] Rode B et al. Piezo1 channels sense whole body physical activity to reset cardiovascular homeostasis and enhance performance. Nat. Commun., 2017, 8

[65] Pathak MM et al. Stretch-activated ion channel Piezo1 directs lineage choice in human neural stem cells. Proc. Natl. Acad. Sci. USA., 2014, 111: 16148-16153

[66] Retailleau K et al. Piezo1 in smooth muscle cells is involved in hypertension-dependent arterial remodeling. Cell Rep., 2015, 13: 1161-1171

[67] Wuertz K et al. In vivo remodeling of intervertebral discs in response to short- and long-term dynamic compression. J. Orthop. Res., 2009, 27: 1235-1242

[68] Gruber HE et al. Vertebral endplate architecture and vascularization: application of micro-computerized tomography, a vascular tracer, and immunocytochemistry in analyses of disc degeneration in the aging sand rat. Spine, 2005, 30: 2593-2600

Funding

National Natural Science Foundation of China (National Science Foundation of China)(81874022)

Key R&D Project of Shandong Province (2022CXGC010503)

Natural Science Foundation of Shandong Province (Shandong Provincial Natural Science Foundation)(ZR202102210113)

Taishan Scholar Project of Shandong Province(tsqn202211317)