CXCR4-targeted nanotoxins induce GSDME-dependent pyroptosis in head and neck squamous cell carcinoma
Rioja Blanco, Elisa (Institut Germans Trias i Pujol. Institut de Recerca contra la Leucèmia Josep Carreras)
Arroyo Solera, Irene (Institut d'Investigació Biomèdica Sant Pau)
Álamo, Patricia (Institut d'Investigació Biomèdica Sant Pau)
Casanova Rigat, Isolda (Institut d'Investigació Biomèdica Sant Pau)
Gallardo, Alberto (Institut d'Investigació Biomèdica Sant Pau)
Unzueta Elorza, Ugutz (Institut d'Investigació Biomèdica Sant Pau)
Serna, Naroa (Universitat Autònoma de Barcelona. Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")
Sánchez-García, Laura (Universitat Autònoma de Barcelona. Departament de Genètica i de Microbiologia)
Quer, Miquel (Institut d'Investigació Biomèdica Sant Pau)
Villaverde Corrales, Antonio (Universitat Autònoma de Barcelona. Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")
Vázquez Gómez, Esther (Universitat Autònoma de Barcelona. Institut de Biotecnologia i de Biomedicina "Vicent Villar Palasí")
León i Vintró, Xavier (Institut d'Investigació Biomèdica Sant Pau)
Alba-Castellón, Lorena (Institut d'Investigació Biomèdica Sant Pau)
Mangues, Ramon 1957- (Institut d'Investigació Biomèdica Sant Pau)

Date:	2022
Abstract:	Therapy resistance, which leads to the development of loco-regional relapses and distant metastases after treatment, constitutes one of the major problems that head and neck squamous cell carcinoma (HNSCC) patients currently face. Thus, novel therapeutic strategies are urgently needed. Targeted drug delivery to the chemokine receptor 4 (CXCR4) represents a promising approach for HNSCC management. In this context, we have developed the self-assembling protein nanotoxins T22-PE24-H6 and T22-DITOX-H6, which incorporate the de-immunized catalytic domain of Pseudomonas aeruginosa (PE24) exotoxin A and the diphtheria exotoxin (DITOX) domain, respectively. Both nanotoxins contain the T22 peptide ligand to specifically target CXCR4-overexpressing HNSCC cells. In this study, we evaluate the potential use of T22-PE24-H6 and T22-DITOX-H6 nanotoxins for the treatment of HNSCC. T22-PE24-H6 and T22-DITOX-H6 CXCR4-dependent cytotoxic effect was evaluated in vitro in two different HNSCC cell lines. Both nanotoxins cell death mechanisms were assessed in HNSCC cell lines by phase-contrast microscopy, AnnexinV/ propidium iodide (PI) staining, lactate dehydrogenase (LDH) release assays, and western blotting. Nanotoxins antitumor effect in vivo was studied in a CXCR4 + HNSCC subcutaneous mouse model. Immunohistochemistry, histopathology, and toxicity analyses were used to evaluate both nanotoxins antitumor effect and possible treatment toxicity. GSMDE and CXCR4 expression in HNSCC patient tumor samples was also assessed by immunohistochemical staining. First, we found that both nanotoxins exhibit a potent CXCR4-dependent cytotoxic effect in vitro. Importantly, nanotoxin treatment triggered caspase-3/Gasdermin E (GSDME)-mediated pyroptosis. The activation of this alternative cell death pathway that differs from traditional apoptosis, becomes a promising strategy to bypass therapy resistance. In addition, T22-PE24-H6 and T22-DITOX-H6 displayed a potent antitumor effect in the absence of systemic toxicity in a CXCR4 + subcutaneous HNSCC mouse model. Lastly, GSDME was found to be overexpressed in tumor tissue from HNSCC patients, highlighting the relevance of this strategy. Altogether, our results show that T22-PE24-H6 and T22-DITOX-H6 represent a promising therapy for HNSCC patients. Remarkably, this is the first study showing that both nanotoxins are capable of activating caspase-3/GSDME-dependent pyroptosis, opening a novel avenue for HNSCC treatment. The online version contains supplementary material available at 10. 1186/s13046-022-02267-8.
Grants:	Instituto de Salud Carlos III PI21/00150 Instituto de Salud Carlos III PI18/00650 Instituto de Salud Carlos III PIE15/00028 Instituto de Salud Carlos III PI15/00378 Instituto de Salud Carlos III PI19/01661 Instituto de Salud Carlos III PI17/00584 Instituto de Salud Carlos III CP19/00028 Agencia Estatal de Investigación BIO2016-76063-R Agencia Estatal de Investigación PID2019-105416RB-I00 Agència de Gestió d'Ajuts Universitaris i de Recerca 2017/SGR-865 Agència de Gestió d'Ajuts Universitaris i de Recerca 2017/SGR-229 Agència de Gestió d'Ajuts Universitaris i de Recerca 2020FI_B2 00168 Agència de Gestió d'Ajuts Universitaris i de Recerca 2018FI_B2_00051 Generalitat de Catalunya PERIS SLT006/17/00093
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Subject:	Targeted drug delivery ; CXCR4 ; HNSCC ; Pyroptosis ; GSDME
Published in:	Journal of Experimental & Clinical Cancer Research : CR, Vol. 41 (february 2022) , ISSN 1756-9966

DOI: 10.1186/s13046-022-02267-8
PMID: 35120582

17 p, 11.1 MB

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Research literature > UAB research groups literature > Research Centres and Groups (research output) > Health sciences and biosciences > Institut d'Investigació en Ciencies de la Salut Germans Trias i Pujol (IGTP) > Josep Carreras Leukaemia Research Institute
Research literature > UAB research groups literature > Research Centres and Groups (research output) > Health sciences and biosciences > Institut de Biotecnologia i de Biomedicina (IBB)
Research literature > UAB research groups literature > Research Centres and Groups (research output) > Health sciences and biosciences > Institut de Recerca Sant Pau
Articles > Research articles
Articles > Published articles