{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T20:50:10Z","timestamp":1786740610297,"version":"build-2736575974"},"reference-count":29,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2026,1,1]],"date-time":"2026-01-01T00:00:00Z","timestamp":1767225600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T00:00:00Z","timestamp":1786665600000},"content-version":"vor","delay-in-days":225,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100001779","name":"Monash University","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100001779","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Mol Neurobiol"],"published-print":{"date-parts":[[2026,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Polymyxins remain indispensable last-line antibiotics for multidrug-resistant Gram-negative infections, yet their clinical use in central nervous system (CNS) infections is constrained by poorly understood neurotoxicity. Here, we define the early molecular signalling events underlying polymyxin B-induced CNS toxicity using an integrated phosphoproteomic and Reverse Phase Protein Array (RPPA) approach in rat brain following intracerebroventricular administration. Global phosphoproteomics revealed extensive phosphosite coverage but identified a highly selective set of significantly regulated phosphosites, implicating calcium-dependent signalling, transcriptional stress regulation, synaptic signalling, and cytoskeletal control, while parallel total proteomics showed minimal changes in protein abundance. RPPA profiling independently confirmed coordinated modulation of stress, apoptotic and survival-associated signalling pathways, including p53, CREB, SQSTM1, Bcl-2, and NF\u03baB related nodes. Network and functional enrichment analyses converged on DNA damage signalling, apoptotic regulation and growth factor-mediated pathways as central features of the polymyxin B early neurotoxicity response, while phosphor to total protein analyses demonstrated suppression of proliferative and pro-survival signalling. Together, these data establish phosphorylation-driven signalling reprogramming as a primary early mechanism of polymyxin B-induced neurotoxicity, providing a mechanistic framework that links membrane-active antibiotic exposure to neuronal stress signalling and identifies candidate pathways for toxicity biomarkers and neuroprotective strategies.<\/jats:p>","DOI":"10.1007\/s12035-026-06126-x","type":"journal-article","created":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T20:24:15Z","timestamp":1786739055000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Systems-Level Phosphoproteomic and RPPA Profiling Reveals Stress and DNA Damage Signalling as Early Drivers of Polymyxin B Neurotoxicity"],"prefix":"10.1007","volume":"63","author":[{"given":"Maytham","family":"Hussein","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thuraya Safaa","family":"Ansaf","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Terry C. C. Lim Kam","family":"Sian","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark","family":"Baker","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pouya","family":"Faridi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhi Ying","family":"Kho","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nivedhitha","family":"Selvakumar","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Keith S.","family":"Kaye","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gauri G.","family":"Rao","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tony","family":"Velkov","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,8,14]]},"reference":[{"issue":"1","key":"6126_CR1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-024-78769-6","volume":"14","author":"M Hussein","year":"2024","unstructured":"Hussein M, Kang Z, Neville SL, Allobawi R, Thrombare V, Koh AJJ et al (2024) Metabolic profiling unveils enhanced antibacterial synergy of polymyxin B and teixobactin against multi-drug resistant Acinetobacter baumannii. Sci Rep 14(1)","journal-title":"Sci Rep"},{"issue":"5","key":"6126_CR2","doi-asserted-by":"publisher","first-page":"1898","DOI":"10.1021\/jm900999h","volume":"53","author":"T Velkov","year":"2010","unstructured":"Velkov T, Thompson PE, Nation RL, Li J (2010) Structure\u2212activity relationships of polymyxin antibiotics. J Med Chem 53(5):1898\u20131916","journal-title":"J Med Chem"},{"issue":"1","key":"6126_CR3","doi-asserted-by":"publisher","DOI":"10.1186\/cc3995","volume":"10","author":"ME Falagas","year":"2006","unstructured":"Falagas ME, Kasiakou SK (2006) Toxicity of polymyxins: a systematic review of the evidence from old and recent studies. Crit Care 10(1)","journal-title":"Crit Care"},{"issue":"6","key":"6126_CR4","doi-asserted-by":"publisher","first-page":"1206","DOI":"10.1093\/jac\/dkm357","volume":"60","author":"AP Zavascki","year":"2007","unstructured":"Zavascki AP, Goldani LZ, Li J, Nation RL (2007) Polymyxin B for the treatment of multidrug-resistant pathogens: a critical review. J Antimicrob Chemother 60(6):1206\u20131215","journal-title":"J Antimicrob Chemother"},{"issue":"6","key":"6126_CR5","doi-asserted-by":"publisher","first-page":"535","DOI":"10.1097\/QCO.0b013e328332e672","volume":"22","author":"RL Nation","year":"2009","unstructured":"Nation RL, Li J (2009) Colistin in the 21st century. Curr Opin Infect Dis 22(6):535\u2013543","journal-title":"Curr Opin Infect Dis"},{"key":"6126_CR6","doi-asserted-by":"publisher","first-page":"305","DOI":"10.1007\/978-3-030-16373-0_18","volume":"1145","author":"MAK Azad","year":"2019","unstructured":"Azad MAK, Nation RL, Velkov T, Li J (2019) Mechanisms of Polymyxin-Induced Nephrotoxicity. Adv Exp Med Biol 1145:305\u2013319","journal-title":"Adv Exp Med Biol"},{"issue":"9","key":"6126_CR7","doi-asserted-by":"publisher","first-page":"589","DOI":"10.1016\/S1473-3099(06)70580-1","volume":"6","author":"J Li","year":"2006","unstructured":"Li J, Nation RL, Turnidge JD, Milne RW, Coulthard K, Rayner CR et al (2006) Colistin: the re-emerging antibiotic for multidrug-resistant Gram-negative bacterial infections. Lancet Infect Dis 6(9):589\u2013601","journal-title":"Lancet Infect Dis"},{"issue":"1","key":"6126_CR8","doi-asserted-by":"publisher","first-page":"66","DOI":"10.1136\/ejhpharm-2023-003786","volume":"31","author":"I Bayraktar","year":"2023","unstructured":"Bayraktar I, Halacli B, Demirkan K, Topeli A (2023) Polymyxin B-related neurotoxicity: a brief case report. Eur J Hosp Pharm 31(1):66\u201367","journal-title":"Eur J Hosp Pharm"},{"issue":"7","key":"6126_CR9","doi-asserted-by":"publisher","first-page":"1021","DOI":"10.1007\/s00134-017-4780-6","volume":"43","author":"T T\u00e4ngd\u00e9n","year":"2017","unstructured":"T\u00e4ngd\u00e9n T, Ramos Mart\u00edn V, Felton TW, Nielsen EI, Marchand S, Br\u00fcggemann RJ et al (2017) The role of infection models and PK\/PD modelling for optimising care of critically ill patients with severe infections. Intensive Care Med 43(7):1021\u20131032","journal-title":"Intensive Care Med"},{"issue":"8","key":"6126_CR10","doi-asserted-by":"publisher","first-page":"4416","DOI":"10.1128\/AAC.00231-12","volume":"56","author":"R Imberti","year":"2012","unstructured":"Imberti R, Cusato M, Accetta G, Marin\u00f2 V, Procaccio F, Gaudio AD et al (2012) Pharmacokinetics of colistin in cerebrospinal fluid after intraventricular administration of colistin methanesulfonate. Antimicrob Agents Chemother 56(8):4416\u20134421","journal-title":"Antimicrob Agents Chemother"},{"issue":"6","key":"6126_CR11","doi-asserted-by":"publisher","first-page":"1333","DOI":"10.1378\/chest.10-0463","volume":"138","author":"R Imberti","year":"2010","unstructured":"Imberti R, Cusato M, Villani P, Carnevale L, Iotti GA, Langer M et al (2010) Steady-state pharmacokinetics and BAL concentration of colistin in critically ill patients after IV colistin methanesulfonate administration. Chest 138(6):1333\u20139","journal-title":"Chest"},{"issue":"1","key":"6126_CR12","doi-asserted-by":"publisher","first-page":"120","DOI":"10.1021\/acschemneuro.8b00300","volume":"10","author":"C Dai","year":"2019","unstructured":"Dai C, Xiao X, Li J, Ciccotosto GD, Cappai R, Tang S et al (2019) Molecular mechanisms of neurotoxicity induced by polymyxins and chemoprevention. ACS Chem Neurosci 10(1):120\u2013131","journal-title":"ACS Chem Neurosci"},{"issue":"3","key":"6126_CR13","doi-asserted-by":"publisher","first-page":"1317","DOI":"10.1007\/s12035-022-03140-7","volume":"60","author":"J Lu","year":"2023","unstructured":"Lu J, Zhu Y, Parkington HC, Hussein M, Zhao J, Bergen P et al (2023) Transcriptomic Mapping of Neurotoxicity Pathways in the Rat Brain in Response to Intraventricular Polymyxin B. Mol Neurobiol 60(3):1317\u20131330","journal-title":"Mol Neurobiol"},{"key":"6126_CR14","doi-asserted-by":"publisher","first-page":"6067","DOI":"10.1016\/j.csbj.2022.10.041","volume":"20","author":"M Hussein","year":"2022","unstructured":"Hussein M, Oberrauch S, Allobawi R, Cornthwaite-Duncan L, Lu J, Sharma R et al (2022) Untargeted metabolomics to evaluate polymyxin B toxicodynamics following direct intracerebroventricular administration into the rat brain. Comput Struct Biotechnol J 20:6067\u201377","journal-title":"Comput Struct Biotechnol J"},{"key":"6126_CR15","first-page":"93624084","volume":"936","author":"P Nazemi","year":"2024","unstructured":"Nazemi P, Talebi F, Emami KH, Jafarieh A, Albaji M, Davoudi-Monfared E (2024) Neurotoxicity of Polymyxins: a Review of the Literature. ESS Open Archive eprints 936:93624084","journal-title":"ESS Open Archive eprints"},{"issue":"4","key":"6126_CR16","doi-asserted-by":"publisher","DOI":"10.1002\/mco2.175","volume":"3","author":"K Pang","year":"2020","unstructured":"Pang K, Wang W, Qin JX, Shi ZD, Hao L, Ma YY et al (2020) Role of protein phosphorylation in cell signaling, disease, and the intervention therapy. MedComm (2020) 3(4)","journal-title":"MedComm (2020)"},{"issue":"23","key":"6126_CR17","doi-asserted-by":"publisher","first-page":"12475","DOI":"10.1073\/pnas.220418997","volume":"97","author":"H-C Huang","year":"2000","unstructured":"Huang H-C, Nguyen T, Pickett CB (2000) Regulation of the antioxidant response element by protein kinase C-mediated phosphorylation of NF-E2-related factor 2. Proc Natl Acad Sci U S A 97(23):12475\u201312480","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"10","key":"6126_CR18","doi-asserted-by":"publisher","first-page":"2512","DOI":"10.1158\/1535-7163.MCT-06-0334","volume":"5","author":"R Tibes","year":"2006","unstructured":"Tibes R, Qiu Y, Lu Y, Hennessy B, Andreeff M, Mills GB et al (2006) Reverse phase protein array: validation of a novel proteomic technology and utility for analysis of primary leukemia specimens and hematopoietic stem cells. Mol Cancer Ther 5(10):2512\u201321","journal-title":"Mol Cancer Ther"},{"key":"6126_CR19","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1016\/j.pharmthera.2017.07.012","volume":"181","author":"T Velkov","year":"2018","unstructured":"Velkov T, Dai C, Ciccotosto GD, Cappai R, Hoyer D, Li J (2018) Polymyxins for CNS infections: pharmacology and neurotoxicity. Pharmacol Ther 181:85\u201390","journal-title":"Pharmacol Ther"},{"issue":"1","key":"6126_CR20","doi-asserted-by":"publisher","DOI":"10.1186\/s12879-024-09759-2","volume":"24","author":"P Aysert-Yildiz","year":"2024","unstructured":"Aysert-Yildiz P, \u00d6Zgen-Top \u00d6, \u015eEnt\u00dcRk AF, Kanik S, \u00d6Zger HS, D\u0130Zbay M (2024) Polymyxin B vs. colistin: the comparison of neurotoxic and nephrotoxic effects of the two polymyxins. BMC Infect Dis 24(1)","journal-title":"BMC Infect Dis"},{"key":"6126_CR21","doi-asserted-by":"publisher","DOI":"10.3389\/fneur.2024.1342419","author":"AX Chin","year":"2024","unstructured":"Chin AX, Ng KW, Chan YC, Goh Y, Rathakrishnan R (2024) Polymyxin-induced neuromuscular weakness: a case report. Front Neurol. https:\/\/doi.org\/10.3389\/fneur.2024.1342419","journal-title":"Front Neurol"},{"issue":"7","key":"6126_CR22","doi-asserted-by":"publisher","first-page":"2259","DOI":"10.1007\/s00204-018-2192-1","volume":"92","author":"B Yun","year":"2018","unstructured":"Yun B, Zhang T, Azad MAK, Wang J, Nowell CJ, Kalitsis P et al (2018) Polymyxin B causes DNA damage in HK-2 cells and mice. Arch Toxicol 92(7):2259\u20132271","journal-title":"Arch Toxicol"},{"issue":"12","key":"6126_CR23","doi-asserted-by":"publisher","first-page":"3394","DOI":"10.1021\/acsinfecdis.5c00782","volume":"11","author":"Z Cai","year":"2025","unstructured":"Cai Z, Zhang Y, Xin Y, Han L, Xu B, Du M et al (2025) Breaking the toxicity-efficacy dilemma of polymyxins: advances in synergists. ACS Infect Dis 11(12):3394\u20133406","journal-title":"ACS Infect Dis"},{"key":"6126_CR24","doi-asserted-by":"publisher","DOI":"10.3389\/fnagi.2021.662474","author":"GY Ou","year":"2021","unstructured":"Ou GY, Lin WW, Zhao WJ (2021) Neuregulins in neurodegenerative diseases. Front Aging Neurosci. https:\/\/doi.org\/10.3389\/fnagi.2021.662474","journal-title":"Front Aging Neurosci"},{"key":"6126_CR25","doi-asserted-by":"publisher","unstructured":"P\u00e9rez-Garc\u00eda P, Mart\u00ednez-G\u00f3mez N, G\u00f3rgolas SV-d, Chamorro-Francisco A, Pardillo-D\u00edaz R, Nunez-Abades P et al (2025) ErbB signaling in brain injury regeneration: pathway interactions and therapeutic potential. Neural Regen Res 21(6):2275\u20132285. https:\/\/doi.org\/10.4103\/NRR.NRR-D-25-00155","DOI":"10.4103\/NRR.NRR-D-25-00155"},{"issue":"3","key":"6126_CR26","first-page":"226","volume":"3","author":"RK Rajan","year":"2025","unstructured":"Rajan RK (2025) A comprehensive review on adaptive plasticity and recovery mechanisms post-acquired brain injury. Neuroprotection (New York NY) 3(3):226\u2013252","journal-title":"Neuroprotection (New York NY)"},{"issue":"2","key":"6126_CR27","doi-asserted-by":"publisher","first-page":"65","DOI":"10.1007\/s11481-014-9534-4","volume":"9","author":"S Buch","year":"2014","unstructured":"Buch S (2014) Growth factor signaling: implications for disease & therapeutics. J Neuroimmune Pharmacol 9(2):65\u201368","journal-title":"J Neuroimmune Pharmacol"},{"key":"6126_CR28","doi-asserted-by":"publisher","unstructured":"Oberrauch S, Lu J, Cornthwaite-Duncan L, Hussein M, Li J, Rao G et al (2022) Intraventricular drug delivery and sampling for pharmacokinetics and pharmacodynamics study. J Vis Exp (181):e63540. https:\/\/doi.org\/10.3791\/63540","DOI":"10.3791\/63540"},{"key":"6126_CR29","doi-asserted-by":"publisher","unstructured":"Lim Kam Sian TCC, Sun C, Cain JE, Steele JR, Hanchapola I, Stoychev S et al (2024) A semiautomated proteomics and phosphoproteomics protocol for the identification of novel therapeutic targets and predictive biomarkers in in vivo xenograft models of pediatric cancers. In: Saad MI (ed) Patient-derived xenografts: methods and protocols. Methods in Molecular Biology, vol 2806. Humana Press, New York, pp 229\u2013242. https:\/\/doi.org\/10.1007\/978-1-0716-3858-3_17","DOI":"10.1007\/978-1-0716-3858-3_17"}],"container-title":["Molecular Neurobiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s12035-026-06126-x.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1007\/s12035-026-06126-x","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1007\/s12035-026-06126-x.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,14]],"date-time":"2026-08-14T20:24:16Z","timestamp":1786739056000},"score":1,"resource":{"primary":{"URL":"https:\/\/link.springer.com\/10.1007\/s12035-026-06126-x"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1]]},"references-count":29,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2026,1]]}},"alternative-id":["6126"],"URL":"https:\/\/doi.org\/10.1007\/s12035-026-06126-x","relation":{},"ISSN":["0893-7648","1559-1182"],"issn-type":[{"value":"0893-7648","type":"print"},{"value":"1559-1182","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1]]},"assertion":[{"value":"6 March 2026","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"5 August 2026","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"14 August 2026","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","label":"Competing Interests","group":{"name":"EthicsHeading","label":"Declarations"}}],"article-number":"835"}}