{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T19:41:22Z","timestamp":1790106082329,"version":"4.0.1"},"reference-count":65,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2025,8,25]],"date-time":"2025-08-25T00:00:00Z","timestamp":1756080000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100000024","name":"Canadian Institutes of Health Research","doi-asserted-by":"publisher","award":["PJT-166062"],"award-info":[{"award-number":["PJT-166062"]}],"id":[{"id":"10.13039\/501100000024","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000024","name":"Canadian Institutes of Health Research","doi-asserted-by":"publisher","award":["FBD-181422"],"award-info":[{"award-number":["FBD-181422"]}],"id":[{"id":"10.13039\/501100000024","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>In breast cancer, progression from localized stage I to distant metastatic stage IV disease is associated with a reduction of 5-year survival from nearly 100% to 23.2%. Expression of the calcium-activated protease isoforms calpain-1 and calpain-2 has been correlated with cell migration and invasion in vitro, metastatic potential in preclinical mouse models of cancer, and breast cancer prognosis in patients. It is unclear which of these two calpain isoforms is responsible for the apparent metastatic potential of cancer cells. Here, we demonstrate that while individual CRISPR-Cas9 knockouts of either CAPN1 or CAPN2 genes (encoding the catalytic subunits of calpain-1 and -2, respectively) reduce in vitro migration and marginally suppress in vivo metastasis, genetic disruption of both calpain-1 and calpain-2 through knockout of the CAPNS1 gene (encoding the common regulatory subunit of calpain-1 and -2) diminishes metastasis by 83.4 \u00b1 13.6% in a mouse xenograft model of human triple-negative breast cancer. The effect of calpain-1\/2 deficiency was replicated in vitro with a modified cell-permeable calpastatin (CAST)-based peptide inhibitor (cell migration reduced to 53.5 \u00b1 11.0% of vehicle control). However, this peptide inhibitor was not effective in vivo at reducing metastasis under the conditions used (vehicle vs. CAST, 1.12 \u00b1 1.35 lung metastases per mm2 vs. 0.34 \u00b1 0.20 metastases per mm2), likely due to rapid clearance, as indicated by the short serum half-life. This work demonstrates that calpain-1\/2 disruption effectively abrogates metastasis and provides rationale for development of effective calpain inhibitors.<\/jats:p>","DOI":"10.3390\/cells14171314","type":"journal-article","created":{"date-parts":[[2025,8,26]],"date-time":"2025-08-26T06:25:57Z","timestamp":1756189557000},"page":"1314","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Calpain-1 and Calpain-2 Promote Breast Cancer Metastasis"],"prefix":"10.3390","volume":"14","author":[{"ORCID":"https:\/\/orcid.org\/0009-0009-1184-4923","authenticated-orcid":false,"given":"Danielle","family":"Harper","sequence":"first","affiliation":[{"name":"Department Pathology and Molecular Medicine, Queen\u2019s University, Kingston, ON K7L3N6, Canada"},{"name":"Division of Cancer Biology and Genetics, Sinclair Cancer Research Institute, Queen\u2019s University, Kingston, ON K7L3N6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jung Yeon","family":"Min","sequence":"additional","affiliation":[{"name":"Department Pathology and Molecular Medicine, Queen\u2019s University, Kingston, ON K7L3N6, Canada"},{"name":"Division of Cancer Biology and Genetics, Sinclair Cancer Research Institute, Queen\u2019s University, Kingston, ON K7L3N6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"James A.","family":"MacLeod","sequence":"additional","affiliation":[{"name":"Department Pathology and Molecular Medicine, Queen\u2019s University, Kingston, ON K7L3N6, Canada"},{"name":"Division of Cancer Biology and Genetics, Sinclair Cancer Research Institute, Queen\u2019s University, Kingston, ON K7L3N6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Samantha","family":"Cockburn","sequence":"additional","affiliation":[{"name":"Department Pathology and Molecular Medicine, Queen\u2019s University, Kingston, ON K7L3N6, Canada"},{"name":"Division of Cancer Biology and Genetics, Sinclair Cancer Research Institute, Queen\u2019s University, Kingston, ON K7L3N6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-0835-833X","authenticated-orcid":false,"given":"Iryna","family":"Predko","sequence":"additional","affiliation":[{"name":"Department Pathology and Molecular Medicine, Queen\u2019s University, Kingston, ON K7L3N6, Canada"},{"name":"Division of Cancer Biology and Genetics, Sinclair Cancer Research Institute, Queen\u2019s University, Kingston, ON K7L3N6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yan","family":"Gao","sequence":"additional","affiliation":[{"name":"Department Pathology and Molecular Medicine, Queen\u2019s University, Kingston, ON K7L3N6, Canada"},{"name":"Division of Cancer Biology and Genetics, Sinclair Cancer Research Institute, Queen\u2019s University, Kingston, ON K7L3N6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter A.","family":"Greer","sequence":"additional","affiliation":[{"name":"Department Pathology and Molecular Medicine, Queen\u2019s University, Kingston, ON K7L3N6, Canada"},{"name":"Division of Cancer Biology and Genetics, Sinclair Cancer Research Institute, Queen\u2019s University, Kingston, ON K7L3N6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-3111-6770","authenticated-orcid":false,"given":"Ivan","family":"Shapovalov","sequence":"additional","affiliation":[{"name":"Department Pathology and Molecular Medicine, Queen\u2019s University, Kingston, ON K7L3N6, Canada"},{"name":"Division of Cancer Biology and Genetics, Sinclair Cancer Research Institute, Queen\u2019s University, Kingston, ON K7L3N6, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,8,25]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1195","DOI":"10.1038\/bjc.2013.6","article-title":"Breast cancer survival and stage at diagnosis in Australia, Canada, Denmark, Norway, Sweden and the UK, 2000\u20132007: A population-based study","volume":"108","author":"Walters","year":"2013","journal-title":"Br. J. Cancer"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1007\/s10549-024-07580-8","article-title":"A Canadian real-world, multi-center, prospective, observational study assessing the treatment duration, the treatment sequence, and the overall survival for patients treated with endocrine therapy +\/- targeted therapy in HR + HER2-negative advanced breast cancer","volume":"210","author":"Doyle","year":"2025","journal-title":"Breast Cancer Res. Treat."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1016\/S1470-2045(24)00757-5","article-title":"Timing of nivolumab with neoadjuvant carboplatin and paclitaxel for early triple-negative breast cancer (BCT1902\/IBCSG 61-20; Neo-N): A non-comparative, open-label, randomised, phase 2 trial","volume":"26","author":"Zdenkowski","year":"2025","journal-title":"Lancet Oncol."},{"key":"ref_4","first-page":"3","article-title":"Five-year cancer survival by stage at diagnosis in Canada","volume":"34","author":"Ellison","year":"2023","journal-title":"Health Rep."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1016\/j.bbapap.2011.08.005","article-title":"Calpains: An elaborate proteolytic system","volume":"1824","author":"Ono","year":"2012","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"854","DOI":"10.1038\/nrd.2016.212","article-title":"Calpain research for drug discovery: Challenges and potential","volume":"15","author":"Ono","year":"2016","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1002\/dvg.20216","article-title":"Conditional disruption of ubiquitous calpains in the mouse","volume":"44","author":"Tan","year":"2006","journal-title":"Genesis"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"364","DOI":"10.1038\/nrc3050","article-title":"The calpain system and cancer","volume":"11","author":"Storr","year":"2011","journal-title":"Nat. Rev. Cancer"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"112220","DOI":"10.1016\/j.yexcr.2020.112220","article-title":"Capn4 contributes to tumor invasion and metastasis in gastric cancer via activation of the Wnt\/beta-catenin\/MMP9 signalling pathways","volume":"395","author":"Zhao","year":"2020","journal-title":"Exp. Cell Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"924","DOI":"10.1007\/s12094-018-02006-6","article-title":"High calpain-1 expression predicts a poor clinical outcome and contributes to tumor progression in pancreatic cancer patients","volume":"21","author":"Yu","year":"2019","journal-title":"Clin. Transl. Oncol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"2422","DOI":"10.1111\/j.1582-4934.2012.01559.x","article-title":"Calpain-2 expression is associated with response to platinum based chemotherapy, progression-free and overall survival in ovarian cancer","volume":"16","author":"Storr","year":"2012","journal-title":"J. Cell Mol. Med."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Tang, S., Yin, Q., Liu, F., and Zhang, Y. (2019). Calpain Small Subunit 1 Protein in the Prognosis of Cancer Survivors and Its Clinicopathological Correlation. Biomed. Res. Int., 2019.","DOI":"10.1155\/2019\/8053706"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"8233","DOI":"10.1002\/jcp.27649","article-title":"Evidence for calpains in cancer metastasis","volume":"234","author":"Chen","year":"2019","journal-title":"J. Cell Physiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"W521","DOI":"10.1093\/nar\/gkae241","article-title":"The EMBL-EBI Job Dispatcher sequence analysis tools framework in 2024","volume":"52","author":"Madeira","year":"2024","journal-title":"Nucleic Acids Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"588","DOI":"10.1073\/pnas.97.2.588","article-title":"The crystal structure of calcium-free human m-calpain suggests an electrostatic switch mechanism for activation by calcium","volume":"97","author":"Strobl","year":"2000","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"493","DOI":"10.1038\/s41586-024-07487-w","article-title":"Accurate structure prediction of biomolecular interactions with AlphaFold 3","volume":"630","author":"Abramson","year":"2024","journal-title":"Nature"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1106","DOI":"10.1111\/j.1600-0854.2012.01371.x","article-title":"Regulated intramembrane cleavage of the EGF receptor","volume":"13","author":"Liao","year":"2012","journal-title":"Traffic"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"1339","DOI":"10.1038\/onc.2009.422","article-title":"Calpain regulates sensitivity to trastuzumab and survival in HER2-positive breast cancer","volume":"29","author":"Kulkarni","year":"2010","journal-title":"Oncogene"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"24435","DOI":"10.1074\/jbc.M203457200","article-title":"Calpain cleaves RhoA generating a dominant-negative form that inhibits integrin-induced actin filament assembly and cell spreading","volume":"277","author":"Kulkarni","year":"2002","journal-title":"J. Biol. Chem."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"977","DOI":"10.1038\/ncb1175","article-title":"Calpain-mediated proteolysis of talin regulates adhesion dynamics","volume":"6","author":"Franco","year":"2004","journal-title":"Nat. Cell Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"11418","DOI":"10.1074\/jbc.M109.090746","article-title":"Regulation of adhesion dynamics by calpain-mediated proteolysis of focal adhesion kinase (FAK)","volume":"285","author":"Chan","year":"2010","journal-title":"J. Biol. Chem."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"5726","DOI":"10.1038\/sj.onc.1209582","article-title":"Calpain 2 and Src dependence distinguishes mesenchymal and amoeboid modes of tumour cell invasion: A link to integrin function","volume":"25","author":"Carragher","year":"2006","journal-title":"Oncogene"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"e70147","DOI":"10.1002\/mco2.70147","article-title":"Calpain 2 Isoform-Specific Cleavage of Filamin A Enhances HIF1\u03b1 Nuclear Translocation, Promoting Metastasis in Triple-Negative Breast Cancer","volume":"6","author":"Jeon","year":"2025","journal-title":"MedComm"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1372","DOI":"10.1074\/jbc.M208772200","article-title":"The role of calpain in the proteolytic cleavage of E-cadherin in prostate and mammary epithelial cells","volume":"278","author":"Day","year":"2003","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"689","DOI":"10.1101\/gad.231894.113","article-title":"Stress-induced cleavage of Myc promotes cancer cell survival","volume":"28","author":"Ngouenet","year":"2014","journal-title":"Genes. Dev."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"53","DOI":"10.1002\/1097-4644(20010101)80:1<53::AID-JCB60>3.0.CO;2-E","article-title":"N-terminal cleavage of bax by calpain generates a potent proapoptotic 18-kDa fragment that promotes bcl-2-independent cytochrome C release and apoptotic cell death","volume":"80","author":"Gao","year":"2000","journal-title":"J. Cell Biochem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"731","DOI":"10.1152\/physrev.00029.2002","article-title":"The calpain system","volume":"83","author":"Goll","year":"2003","journal-title":"Physiol. Rev."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"7079604","DOI":"10.1155\/2019\/7079604","article-title":"The Role of Nonerythroid Spectrin \u03b1II in Cancer","volume":"2019","author":"Ackermann","year":"2019","journal-title":"J. Oncol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"783","DOI":"10.1038\/nmeth.3047","article-title":"Improved vectors and genome-wide libraries for CRISPR screening","volume":"11","author":"Sanjana","year":"2014","journal-title":"Nat. Methods"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1006\/abio.2001.5606","article-title":"Casein zymography of calpains using a 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid-imidazole buffer","volume":"304","author":"Croall","year":"2002","journal-title":"Anal. Biochem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1931","DOI":"10.1038\/nprot.2014.131","article-title":"Quantitative and unbiased analysis of directional persistence in cell migration","volume":"9","author":"Gorelik","year":"2014","journal-title":"Nat. Protoc."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2078","DOI":"10.1128\/MCB.01062-15","article-title":"Calpain Genetic Disruption and HSP90 Inhibition Combine To Attenuate Mammary Tumorigenesis","volume":"36","author":"Grieve","year":"2016","journal-title":"Mol. Cell Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2761","DOI":"10.4049\/jimmunol.162.5.2761","article-title":"Dissecting NK cell development using a novel alymphoid mouse model: Investigating the role of the c-abl proto-oncogene in murine NK cell differentiation","volume":"162","author":"Colucci","year":"1999","journal-title":"J. Immunol."},{"key":"ref_34","unstructured":"O\u2019Clair, L., Shean, J., and Kolozsvari, B. (2017). Live-Cell Analysis Handbook: A Guide to Real-Time Live-Cell Imaging and Analysis, Sartorius, Essen BioScience, Inc.. [6th ed.]."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1080\/14728222.2022.2047178","article-title":"Calpain as a therapeutic target in cancer","volume":"26","author":"Shapovalov","year":"2022","journal-title":"Expert. Opin. Ther. Targets"},{"key":"ref_36","unstructured":"Harper, D., Min, J.Y., MacLeod, J.A., Cockburn, S., Predko, I., Gao, Y., Greer, P., and Shapovalov, I. (2025, June 20). Dataset Related to Article: \u201cCalpain-1 and Calpain-2 Promote Breast Cancer Metastasis\u201d (Version 1). Zenodo. Available online: https:\/\/zenodo.org\/records\/16790974."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"409","DOI":"10.1038\/nature07451","article-title":"Calcium-bound structure of calpain and its mechanism of inhibition by calpastatin","volume":"456","author":"Hanna","year":"2008","journal-title":"Nature"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1515\/BC.2008.004","article-title":"Inhibition of human mu-calpain by conformationally constrained calpastatin peptides","volume":"389","author":"Pfizer","year":"2008","journal-title":"Biol. Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"10141","DOI":"10.7150\/thno.46985","article-title":"The peptide PROTAC modality: A novel strategy for targeted protein ubiquitination","volume":"10","author":"Jin","year":"2020","journal-title":"Theranostics"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"221","DOI":"10.1016\/j.bbamcr.2017.10.013","article-title":"An easy-to-use FRET protein substrate to detect calpain cleavage in vitro and in vivo","volume":"1865","author":"McCartney","year":"2018","journal-title":"Biochim. Biophys. Acta Mol. Cell Res."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"32719","DOI":"10.1074\/jbc.272.52.32719","article-title":"Regulation of cell migration by the calcium-dependent protease calpain","volume":"272","author":"Huttenlocher","year":"1997","journal-title":"J. Biol. Chem."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1634\/theoncologist.10-90003-20","article-title":"Extending survival with chemotherapy in metastatic breast cancer","volume":"10","year":"2005","journal-title":"Oncologist"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"eaan0026","DOI":"10.1126\/scitranslmed.aan0026","article-title":"Neoadjuvant chemotherapy induces breast cancer metastasis through a TMEM-mediated mechanism","volume":"9","author":"Karagiannis","year":"2017","journal-title":"Sci. Transl. Med."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"456","DOI":"10.1158\/2767-9764.CRC-21-0117","article-title":"Targeting the Ezrin Adaptor Protein Sensitizes Metastatic Breast Cancer Cells to Chemotherapy and Reduces Neoadjuvant Therapy-induced Metastasis","volume":"2","author":"Hoskin","year":"2022","journal-title":"Cancer Res. Commun."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Volmer, L., Koch, A., Matovina, S., Dannehl, D., Weiss, M., Welker, G., Hahn, M., Engler, T., Wallwiener, M., and Walter, C.B. (2022). Neoadjuvant Chemotherapy of Patients with Early Breast Cancer Is Associated with Increased Detection of Disseminated Tumor Cells in the Bone Marrow. Cancers, 14.","DOI":"10.3390\/cancers14030635"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"101330","DOI":"10.1016\/j.xcrm.2023.101330","article-title":"Needle biopsy accelerates pro-metastatic changes and systemic dissemination in breast cancer: Implications for mortality by surgery delay","volume":"4","author":"Kameyama","year":"2023","journal-title":"Cell Rep. Med."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"682","DOI":"10.1002\/pros.23512","article-title":"Calpain-2 triggers prostate cancer metastasis via enhancing CRMP4 promoter methylation through NF-kappaB\/DNMT1 signaling pathway","volume":"78","author":"Gao","year":"2018","journal-title":"Prostate"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"9696","DOI":"10.1074\/jbc.M112.419713","article-title":"A truncated fragment of Src protein kinase generated by calpain-mediated cleavage is a mediator of neuronal death in excitotoxicity","volume":"288","author":"Hossain","year":"2013","journal-title":"J. Biol. Chem."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"957","DOI":"10.1083\/jcb.200708048","article-title":"Calpain 2 and PTP1B function in a novel pathway with Src to regulate invadopodia dynamics and breast cancer cell invasion","volume":"180","author":"Cortesio","year":"2008","journal-title":"J. Cell Biol."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"1862","DOI":"10.1002\/acn3.51169","article-title":"Novel CAPN1 mutations extend the phenotypic heterogeneity in combined spastic paraplegia and ataxia","volume":"7","author":"Lai","year":"2020","journal-title":"Ann. Clin. Transl. Neurol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"781","DOI":"10.1007\/s12311-019-01038-0","article-title":"Increased Diagnostic Yield of Spastic Paraplegia with or Without Cerebellar Ataxia Through Whole-Genome Sequencing","volume":"18","author":"Kim","year":"2019","journal-title":"Cerebellum"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/j.celrep.2016.05.044","article-title":"Defects in the CAPN1 Gene Result in Alterations in Cerebellar Development and Cerebellar Ataxia in Mice and Humans","volume":"16","author":"Wang","year":"2016","journal-title":"Cell Rep."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"C1226","DOI":"10.1152\/ajpcell.00408.2019","article-title":"Loss of calpains-1 and -2 prevents repair of plasma membrane scrape injuries, but not small pores, and induces a severe muscular dystrophy","volume":"318","author":"Piper","year":"2020","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"4097","DOI":"10.1128\/MCB.05189-11","article-title":"Vital Role of the Calpain-Calpastatin System for Placental-Integrity-Dependent Embryonic Survival","volume":"31","author":"Takano","year":"2011","journal-title":"Mol. Cell. Biol."},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Dutt, P., Croall, D.E., Arthur, J.S., Veyra, T.D., Williams, K., Elce, J.S., and Greer, P.A. (2006). m-Calpain is required for preimplantation embryonic development in mice. BMC Dev. Biol., 6.","DOI":"10.1186\/1471-213X-6-3"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1080\/15216540050176610","article-title":"The calpain small subunit gene is essential: Its inactivation results in embryonic lethality","volume":"50","author":"Zimmerman","year":"2000","journal-title":"IUBMB Life"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"21002","DOI":"10.1074\/jbc.M710354200","article-title":"In vivo targeted deletion of calpain small subunit, Capn4, in cells of the osteoblast lineage impairs cell proliferation, differentiation, and bone formation","volume":"283","author":"Shimada","year":"2008","journal-title":"J. Biol. Chem."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"5773","DOI":"10.1523\/JNEUROSCI.4247-12.2013","article-title":"Conditional disruption of calpain in the CNS alters dendrite morphology, impairs LTP, and promotes neuronal survival following injury","volume":"33","author":"Amini","year":"2013","journal-title":"J. Neurosci."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"2799","DOI":"10.1128\/MCB.00157-10","article-title":"Targeted deletion of Capn4 in cells of the chondrocyte lineage impairs chondrocyte proliferation and differentiation","volume":"30","author":"Kashiwagi","year":"2010","journal-title":"Mol. Cell Biol."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"255","DOI":"10.2337\/db15-0963","article-title":"Mitochondrial Calpain-1 Disrupts ATP Synthase and Induces Superoxide Generation in Type 1 Diabetic Hearts: A Novel Mechanism Contributing to Diabetic Cardiomyopathy","volume":"65","author":"Ni","year":"2016","journal-title":"Diabetes"},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Wernimont, S.A., Simonson, W.T., Greer, P.A., Seroogy, C.M., and Huttenlocher, A. (2010). Calpain 4 is not necessary for LFA-1-mediated function in CD4+ T cells. PLoS ONE, 5.","DOI":"10.1371\/journal.pone.0010513"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"L145","DOI":"10.1152\/ajplung.00348.2015","article-title":"Activation of calpain by renin-angiotensin system in pleural mesothelial cells mediates tuberculous pleural fibrosis","volume":"311","author":"Yang","year":"2016","journal-title":"Am. J. Physiol. Lung Cell Mol. Physiol."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"465","DOI":"10.1515\/BC.2004.054","article-title":"Interaction of calpastatin with calpain: A review","volume":"385","author":"Wendt","year":"2004","journal-title":"Biol. Chem."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"1775","DOI":"10.1016\/S0006-2952(99)00259-2","article-title":"Effect of D-amino acid substitution on the stability, the secondary structure, and the activity of membrane-active peptide","volume":"58","author":"Hong","year":"1999","journal-title":"Biochem. Pharmacol."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"855","DOI":"10.1007\/s40262-013-0079-0","article-title":"Pharmacokinetics and pharmacokinetic-pharmacodynamic correlations of therapeutic peptides","volume":"52","author":"Diao","year":"2013","journal-title":"Clin. Pharmacokinet."}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/14\/17\/1314\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:32:26Z","timestamp":1760034746000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/14\/17\/1314"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,8,25]]},"references-count":65,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["cells14171314"],"URL":"https:\/\/doi.org\/10.3390\/cells14171314","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,8,25]]}}}