{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T18:19:35Z","timestamp":1785435575459,"version":"3.56.0"},"reference-count":45,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T00:00:00Z","timestamp":1783036800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81801241"],"award-info":[{"award-number":["81801241"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"award":["81801241"],"award-info":[{"award-number":["81801241"]}],"id":[{"id":"https:\/\/ror.org\/01h0zpd94","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Sichuan Science and Technology Program","award":["2024NSFSC0612"],"award-info":[{"award-number":["2024NSFSC0612"]}]},{"name":"Natural Science Research Project of Anhui Educational Committee","award":["2025AHGXZK31598"],"award-info":[{"award-number":["2025AHGXZK31598"]}]},{"name":"Luzhou Science and Technology Program","award":["2023SYF104"],"award-info":[{"award-number":["2023SYF104"]}]},{"name":"National College Students' innovation and entrepreneurship training program","award":["202510367044"],"award-info":[{"award-number":["202510367044"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>The small heat shock protein HSPB1 is a ubiquitously expressed mechanoresponsive chaperone essential for cytoskeletal remodeling under mechanical load. Mutations in HSPB1, including S135F, cause Charcot-Marie-Tooth (CMT) peripheral neuropathy, yet the mechanisms underlying the selective vulnerability of peripheral nerves remain enigmatic. Here we demonstrate that substrate stiffness is a critical determinant of HSPB1S135F-mediated neurodegeneration. Using stiffness-tunable polydimethylsiloxane (PDMS) substrates (1 kPa, 10 kPa, 2 MPa) and uniaxial cyclic stretch, we show that primary dorsal root ganglia (DRG) neurons and SH-SY5Y cells expressing HSPB1S135F exhibit profound deficits in mechanoadaptation. On compliant substrates (10 kPa), HSPB1S135F causes stretch-induced axon fragmentation and neuronal death, whereas HSPB1WT confers robust neuroprotection. HSPB1S135F also disrupts stiffness-directed neuritogenesis in differentiated SH-SY5Y cells: HSPB1WT-expressing cells show optimal axonal outgrowth and \u03b2III-tubulin expression on 10 kPa substrates mimicking muscle tissue stiffness, while HSPB1S135F mutants display disorganized focal adhesions and complete differentiation failure. Mechanistically, we uncover that HSPB1S135F dysregulates stage-specific transglutaminase (TGase) expression\u2014insufficient TGase during early neuritogenesis impairs filopodia stabilization, whereas aberrant TGase persistence at late stages constrains axon extension. Our findings establish HSPB1 as a biomechanical sensor that integrates ECM stiffness signals to coordinate peripheral nerve regeneration, and identify defective mechanoadaptation as a previously unrecognized pathomechanism in CMT. These results open new avenues for stiffness-targeted therapeutic strategies in peripheral neuropathy.<\/jats:p>","DOI":"10.3390\/cells15131216","type":"journal-article","created":{"date-parts":[[2026,7,3]],"date-time":"2026-07-03T15:15:04Z","timestamp":1783091704000},"page":"1216","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Neuropathy-Associated HSPB1 Mutant Impairs Neuronal Mechanoadaptation and Axonal Regeneration"],"prefix":"10.3390","volume":"15","author":[{"given":"Jiming","family":"Xie","sequence":"first","affiliation":[{"name":"School of Basic Medical Science, Bengbu Medical University, Bengbu 233030, China"},{"name":"School of Basic Medical Science, Southwest Medical University, Luzhou 646000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ronglin","family":"Han","sequence":"additional","affiliation":[{"name":"School of Basic Medical Science, Southwest Medical University, Luzhou 646000, China"},{"name":"Department of Pathology, West China Hospital, Sichuan University, Chengdu 610041, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Haidong","family":"Xu","sequence":"additional","affiliation":[{"name":"School of Basic Medical Science, Bengbu Medical University, Bengbu 233030, China"},{"name":"Key Laboratory of Basic and Clinical Cardiovascular Diseases, Bengbu Medical University, Bengbu 233030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhiyu","family":"Li","sequence":"additional","affiliation":[{"name":"School of Basic Medical Science, Southwest Medical University, Luzhou 646000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jingyi","family":"Zhao","sequence":"additional","affiliation":[{"name":"School of Basic Medical Science, Bengbu Medical University, Bengbu 233030, China"},{"name":"Key Laboratory of Basic and Clinical Cardiovascular Diseases, Bengbu Medical University, Bengbu 233030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5909-2892","authenticated-orcid":false,"given":"Ying","family":"Wan","sequence":"additional","affiliation":[{"name":"School of Basic Medical Science, Southwest Medical University, Luzhou 646000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xianchao","family":"Pan","sequence":"additional","affiliation":[{"name":"School of Basic Medical Science, Bengbu Medical University, Bengbu 233030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juan","family":"Xing","sequence":"additional","affiliation":[{"name":"School of Basic Medical Science, Bengbu Medical University, Bengbu 233030, China"},{"name":"School of Basic Medical Science, Southwest Medical University, Luzhou 646000, China"},{"name":"Key Laboratory of Basic and Clinical Cardiovascular Diseases, Bengbu Medical University, Bengbu 233030, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,3]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"728","DOI":"10.1038\/nrm.2017.108","article-title":"Mechanical forces direct stem cell behaviour in development and regeneration","volume":"18","author":"Vining","year":"2017","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"88","DOI":"10.1016\/j.plrev.2017.06.016","article-title":"Cellular mechanosensing of the biophysical microenvironment: A review of mathematical models of biophysical regulation of cell responses","volume":"22\u201323","author":"Cheng","year":"2017","journal-title":"Phys. Life Rev."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1364","DOI":"10.4103\/NRR.NRR-D-23-01874","article-title":"Stiffness-tunable biomaterials provide a good extracellular matrix environment for axon growth and regeneration","volume":"20","author":"Han","year":"2025","journal-title":"Neural Regen. Res."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"4993","DOI":"10.7150\/thno.74571","article-title":"Biomechanical microenvironment in peripheral nerve regeneration: From pathophysiological understanding to tissue","volume":"12","author":"Kong","year":"2022","journal-title":"Theranostics"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2856","DOI":"10.1038\/s41598-024-52992-7","article-title":"Mechanical activation and expression of HSP27 in epithelial ovarian cancer","volume":"14","author":"Buckley","year":"2024","journal-title":"Sci. Rep."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2661","DOI":"10.1091\/mbc.e17-02-0087","article-title":"Mechanical signals activate p38 MAPK pathway-dependent reinforcement of actin via mechanosensitive HspB1","volume":"28","author":"Hoffman","year":"2017","journal-title":"Mol. Biol. Cell"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"ar100","DOI":"10.1091\/mbc.E22-02-0057","article-title":"Phosphorylation of the small heat shock protein HspB1 regulates cytoskeletal recruitment and cell motility","volume":"33","author":"Hoffman","year":"2022","journal-title":"Mol. Biol. Cell"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"119","DOI":"10.1007\/s12031-010-9481-3","article-title":"Distinct expression of c-Jun and HSP27 in axotomized and spared bulbospinal neurons after cervical spinal cord injury","volume":"45","author":"Vinit","year":"2011","journal-title":"J. Mol. Neurosci."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"602","DOI":"10.1038\/ng1354","article-title":"Mutant small heat-shock protein 27 causes axonal Charcot-Marie-Tooth disease and distal hereditary motor neuropathy","volume":"36","author":"Evgrafov","year":"2004","journal-title":"Nat. Genet."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1111\/jns.12395","article-title":"A novel HSPB1 mutation associated with a late onset CMT2 phenotype: Case presentation and systematic review of the literature","volume":"25","author":"Taga","year":"2020","journal-title":"J. Peripher. Nerv. Syst."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1186\/s12984-023-01172-1","article-title":"Quantitative assessment of plantar pressure patterns in relation to foot deformities in people with hereditary motor and sensory neuropathies","volume":"20","author":"Bloks","year":"2023","journal-title":"J. Neuroeng. Rehabil."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"15320","DOI":"10.1523\/JNEUROSCI.3266-11.2011","article-title":"Small heat-shock protein HSPB1 mutants stabilize microtubules in Charcot-Marie-Tooth neuropathy","volume":"31","author":"Asselbergh","year":"2011","journal-title":"J. Neurosci. Off. J. Soc. Neurosci."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1051","DOI":"10.1080\/15548627.2019.1569930","article-title":"Neuropathy-causing mutations in HSPB1 impair autophagy by disturbing the formation of SQSTM1\/p62 bodies","volume":"15","author":"Haidar","year":"2019","journal-title":"Autophagy"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Zhang, X., Qiao, Y., Han, R., Gao, Y., Yang, X., Zhang, Y., Wan, Y., Yu, W., Pan, X., and Xing, J. (2022). A Charcot-Marie-Tooth-Causing Mutation in HSPB1 Decreases Cell Adaptation to Repeated Stress by Disrupting Autophagic Clearance of Misfolded Proteins. Cells, 11.","DOI":"10.3390\/cells11182886"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"300","DOI":"10.1016\/j.nbd.2017.02.009","article-title":"Defective axonal transport: A common pathological mechanism in inherited and acquired peripheral neuropathies","volume":"105","author":"Prior","year":"2017","journal-title":"Neurobiol. Dis."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"342","DOI":"10.1016\/j.neuron.2023.10.005","article-title":"Mechanics in the nervous system From development to disease","volume":"112","author":"Pillai","year":"2024","journal-title":"Neuron"},{"key":"ref_17","first-page":"839","article-title":"Mechanobiology of peripheral nerve development: Mechanical properties, sensing, and response","volume":"40","author":"Liu","year":"2026","journal-title":"Chin. J. Reparative Reconstr. Surg."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"677","DOI":"10.1016\/j.cell.2006.06.044","article-title":"Matrix elasticity directs stem cell lineage specification","volume":"126","author":"Engler","year":"2006","journal-title":"Cell"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1016\/j.biomaterials.2018.01.036","article-title":"Adsorption force of fibronectin controls transmission of cell traction force and subsequent stem cell fate","volume":"162","author":"Lin","year":"2018","journal-title":"Biomaterials"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"183","DOI":"10.3233\/JND-150144","article-title":"Characterization of New Transgenic Mouse Models for Two Charcot-Marie-Tooth-Causing HspB1 Mutations using the Rosa26 Locus","volume":"3","author":"Bouhy","year":"2016","journal-title":"J. Neuromuscul. Dis."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"968","DOI":"10.1038\/nm.2396","article-title":"HDAC6 inhibitors reverse axonal loss in a mouse model of mutant HSPB1-induced Charcot-Marie-Tooth disease","volume":"17","author":"Krishnan","year":"2011","journal-title":"Nat. Med."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"307","DOI":"10.1016\/j.yexcr.2005.10.007","article-title":"Integration of topographical and biochemical cues by axons during growth on microfabricated 3-D substrates","volume":"311","author":"Li","year":"2005","journal-title":"Exp. Cell Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1002\/jbm.a.37443","article-title":"Enhanced neuronal differentiation by dynamic piezoelectric stimulation","volume":"111","author":"Pinho","year":"2023","journal-title":"J. Biomed. Mater. Res. A"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1016\/j.neuro.2008.11.001","article-title":"Effects of all-trans-retinoic acid on human SH-SY5Y neuroblastoma as in vitro model in neurotoxicity research","volume":"30","author":"Cheung","year":"2009","journal-title":"Neurotoxicology"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"8","DOI":"10.1016\/j.abb.2011.04.016","article-title":"A novel mechanical loading model for studying the distributions of strain and mechano-growth factor expression","volume":"511","author":"Peng","year":"2011","journal-title":"Arch. Biochem. Biophys."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"147","DOI":"10.1007\/s11010-012-1377-9","article-title":"The nuclear localization of MGF receptor in osteoblasts under mechanical stimulation","volume":"369","author":"Peng","year":"2012","journal-title":"Mol. Cell. Biochem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"744","DOI":"10.1002\/jnr.24408","article-title":"Ion current and action potential alterations in peripheral neurons subject to uniaxial strain","volume":"97","author":"Bianchi","year":"2019","journal-title":"J. Neurosci. Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1016\/j.colsurfb.2016.06.044","article-title":"Stretching-induced nanostructures on shape memory polyurethane films and their regulation to osteoblasts morphology","volume":"146","author":"Xing","year":"2016","journal-title":"Colloids Surf. B"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"194","DOI":"10.1016\/j.brainresbull.2018.12.001","article-title":"Computing neurite outgrowth and arborization in superior cervical ganglion neurons","volume":"144","author":"Henley","year":"2019","journal-title":"Brain Res. Bull."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Jocher, G., Mannschatz, S.H., Offterdinger, M., and Schweigreiter, R. (2018). Microfluidics of Small-Population Neurons Allows for a Precise Quantification of the Peripheral Axonal Growth State. Front. Cell Neurosci., 12.","DOI":"10.3389\/fncel.2018.00166"},{"key":"ref_31","first-page":"1401","article-title":"Mechanical breaking of microtubules in axons during dynamic stretch injury underlies delayed elasticity, microtubule disassembly, and axon degeneration","volume":"24","author":"Patel","year":"2010","journal-title":"FASEB J. Off. Publ. Fed. Am. Soc. Exp. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"7423","DOI":"10.1021\/acs.langmuir.8b02003","article-title":"Directing Neuronal Outgrowth and Network Formation of Rat Cortical Neurons by Cyclic Substrate Stretch","volume":"35","author":"Abraham","year":"2019","journal-title":"Langmuir ACS J. Surf. Colloids"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"128","DOI":"10.1016\/j.jmbbm.2019.01.008","article-title":"Shape recovery strain and nanostructures on recovered polyurethane films and their regulation to osteoblasts morphology","volume":"92","author":"Xing","year":"2019","journal-title":"J. Mech. Behav. Biomed. Mater."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Lin, J., Li, X., Yin, J., and Qian, J. (2020). Effect of Cyclic Stretch on Neuron Reorientation and Axon Outgrowth. Front. Bioeng. Biotechnol., 8.","DOI":"10.3389\/fbioe.2020.597867"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"146485","DOI":"10.1016\/j.ijbiomac.2025.146485","article-title":"Research progress on stiffness controllable scaffolds based on gelatin methacryloyl hydrogels for tissue repair and reconstruction","volume":"321","author":"Liu","year":"2025","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"21005","DOI":"10.1002\/jcp.28704","article-title":"Treatment of human neuroblastoma cell line SH-SY5Y with HSP27 siRNA tagged-exosomes decreased differentiation rate into mature neurons","volume":"234","author":"Shokrollahi","year":"2019","journal-title":"J. Cell. Physiol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"10670","DOI":"10.1007\/s12035-025-04888-4","article-title":"Bexarotene Promotes Neuroblastoma SH-SY5Y Cell Differentiation to Mature Neurons with Decreased Proliferation","volume":"62","author":"Zhang","year":"2025","journal-title":"Mol. Neurobiol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"9497","DOI":"10.1038\/s41467-024-53859-1","article-title":"Talin and vinculin combine their activities to trigger actin assembly","volume":"15","author":"Wang","year":"2024","journal-title":"Nat. Commun."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2413","DOI":"10.1093\/emboj\/20.10.2413","article-title":"Role of transglutaminase II in retinoic acid-induced activation of RhoA-associated kinase-2","volume":"20","author":"Singh","year":"2001","journal-title":"EMBO J."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1038\/s42003-026-09661-4","article-title":"Molecular resilience of neurons to repetitive mechanical compression","volume":"9","author":"Coppini","year":"2026","journal-title":"Commun. Biol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1263","DOI":"10.1159\/000485485","article-title":"Mechanosensitivity of Embryonic Neurites Promotes Their Directional Extension and Schwann Cells Progenitors Migration","volume":"44","author":"Rosso","year":"2017","journal-title":"Cell. Physiol. Biochem."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"688","DOI":"10.1038\/s41598-017-18874-x","article-title":"Characterization of human small heat shock protein HSPB1 \u03b1-crystallin domain localized mutants associated with hereditary motor neuron diseases","volume":"8","author":"Weeks","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1007\/s00726-011-1083-z","article-title":"Transglutaminase 2 interaction with small heat shock proteins mediate cell survival upon excitotoxic stress","volume":"44","author":"Caccamo","year":"2013","journal-title":"Amino Acids"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"391","DOI":"10.1074\/jbc.M206361200","article-title":"Tissue transglutaminase mediates activation of RhoA and MAP kinase pathways during retinoic acid-induced neuronal differentiation of SH-SY5Y cells","volume":"278","author":"Singh","year":"2003","journal-title":"J. Biol. Chem."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Schmidt, S.I., Blaabjerg, M., Freude, K., and Meyer, M. (2022). RhoA Signaling in Neurodegenerative Diseases. Cells, 11.","DOI":"10.3390\/cells11091520"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/15\/13\/1216\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,8]],"date-time":"2026-07-08T04:36:47Z","timestamp":1783485407000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/15\/13\/1216"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,3]]},"references-count":45,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2026,7]]}},"alternative-id":["cells15131216"],"URL":"https:\/\/doi.org\/10.3390\/cells15131216","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,3]]}}}