GSDMD-dependent pyroptotic induction by a multivalent CXCR4-targeted nanotoxin blocks colorectal cancer metastases

Drug Deliv. 2022 Dec;29(1):1384-1397. doi: 10.1080/10717544.2022.2069302.

Abstract

Colorectal cancer (CRC) remains the third cause of cancer-related mortality in Western countries, metastases are the main cause of death. CRC treatment remains limited by systemic toxicity and chemotherapy resistance. Therefore, nanoparticle-mediated delivery of cytotoxic agents selectively to cancer cells represents an efficient strategy to increase the therapeutic index and overcome drug resistance. We have developed the T22-PE24-H6 therapeutic protein-only nanoparticle that incorporates the exotoxin A from Pseudomonas aeruginosa to selectively target CRC cells because of its multivalent ligand display that triggers a high selectivity interaction with the CXCR4 receptor overexpressed on the surface of CRC stem cells. We here observed a CXCR4-dependent cytotoxic effect for T22-PE24-H6, which was not mediated by apoptosis, but instead capable of inducing a time-dependent and sequential activation of pyroptotic markers in CRC cells in vitro. Next, we demonstrated that repeated doses of T22-PE24-H6 inhibit tumor growth in a subcutaneous CXCR4+ CRC model, also through pyroptotic activation. Most importantly, this nanoparticle also blocked the development of lymphatic and hematogenous metastases, in a highly aggressive CXCR4+ SW1417 orthotopic CRC model, in the absence of systemic toxicity. This targeted drug delivery approach supports for the first time the clinical relevance of inducing GSDMD-dependent pyroptosis, a cell death mechanism alternative to apoptosis, in CRC models, leading to the selective elimination of CXCR4+ cancer stem cells, which are associated with resistance, metastases and anti-apoptotic upregulation.

Keywords: CXCR4; GSDMD; Metastasis; PE24 exotoxin; colorectal cancer; pyroptosis; targeted nanoparticle.

MeSH terms

  • Antineoplastic Agents* / pharmacology
  • Colorectal Neoplasms* / drug therapy
  • Drug Delivery Systems
  • Humans
  • Neoplasm Metastasis / prevention & control
  • Phosphate-Binding Proteins* / metabolism
  • Pore Forming Cytotoxic Proteins* / metabolism
  • Pyroptosis*
  • Receptors, CXCR4* / metabolism
  • Receptors, CXCR4* / therapeutic use
  • Signal Transduction

Substances

  • Antineoplastic Agents
  • CXCR4 protein, human
  • GSDMD protein, human
  • Phosphate-Binding Proteins
  • Pore Forming Cytotoxic Proteins
  • Receptors, CXCR4

Grants and funding

This work was supported by Instituto de Salud Carlos III (ISCIII, Co-funding from FEDER) [FI16/00017 to R.S., PI18/00650, PIE15/00028, PI21/00150 and EU COST Action CA 17140 to R.M.]; Agencia Estatal de Investigación (AEI) and Fondo Europeo de Desarrollo Regional (FEDER) [grant BIO2016-76063-R, AEI/FEDER, UE to A.V. and PID2019-105416RB-I00/AEI/10.13039/501100011033 to E.V]; CIBER-BBN [CB06/01/1031 and 4NanoMets to R.M., VENOM4CANCER to A.V. NANOSCAPE to U.U. and NANOREMOTE to E.V.]; AGAUR [2017- SGR-865 to R.M., 2017SGR-229 to A.V., and 2018FI_B2_00051 to L.S.G.] and a predoctoral fellowship from the Government of Navarra to N.S.; Josep Carreras Leukemia Research Institute [P/AG to R.M.] and the Generalitat de Catalunya CERCA Programme. A.V. received an Icrea Academia Award. UU was supported by Miguel Servet contract (CP19/00028) from ISCIII co-funded by European Social Fund (ESF investing in your future). The authors would like to thank the ICTS-141007 Nanbiosis Platform, especifically the Platform of Protein Production and the ICTS Nanbiosis Nantoxicology Unit of CIBER-BBN where the bioluminescent follow-up of cancer cells and toxicity studies were performed.