LGR5 as a Therapeutic Target of Antibody-Functionalized Biomimetic Magnetoliposomes for Colon Cancer Therapy

Int J Nanomedicine. 2024 Feb 23:19:1843-1865. doi: 10.2147/IJN.S440881. eCollection 2024.

Abstract

Purpose: The lack of specificity of conventional chemotherapy is one of the main difficulties to be solved in cancer therapy. Biomimetic magnetoliposomes are successful chemotherapy controlled-release systems, hyperthermia, and active targeting agents by functionalization of their surface with monoclonal antibodies. The membrane receptor Leucine-rich repeat-containing G-protein coupled receptor 5 (LGR5) stands out as colorectal cancer (CRC) biomarker and appears to be related to treatment resistance and the development of metastasis. The aim of this study was to assess the effectiveness and safety of LGR5-targeted biomimetic magnetoliposomes loaded with oxaliplatin (OXA) or 5-fluorouracil (5-FU) in the selective treatment of CRC and their possible application in hyperthermia.

Methods: Synthesis, characterization and determination of heating capacity of magnetoliposomes transporting OXA or 5-FU (with and without LGR5 functionalization) were conducted. In vitro antitumoral activity was assayed in multiple colorectal cell lines at different times of exposition. In addition to this, cell internalization was studied by Prussian Blue staining, flow cytometry and fluorescence microscopy. In vivo acute toxicity of magnetoliposomes was performed to evaluate iron-related toxicity.

Results: OXA and 5-FU loaded magnetoliposomes functionalized with LGR5 antibody showed higher cellular uptake than non-targeted nanoformulation with a reduction of the percentage of proliferation in colon cancer cell lines up to 3.2-fold of the IC50 value compared to that of free drug. The differences between non-targeted and targeted nanoformulations were more evident after short exposure times (4 and 8 hours). Interestingly, assays in the MC38 transduced cells with reduced LGR5 expression (MC38-L(-)), showed lower cell internalization of LGR5-targeted magnetoliposomes compared to non-transduced MC38 cell line. In addition, magnetoliposomes showed an in vitro favorable heating response under magnetic excitation and great iron-related biocompatibility data in vivo.

Conclusion: Drug-loaded magnetoliposomes functionalized with anti-LGR5 antibodies could be a promising CRC treatment strategy for LGR5+ targeted chemotherapy, magnetic hyperthermia, and both in combination.

Keywords: 5-fluorouracil; LGR5; colorectal neoplasms; magnetic hyperthermia; magnetoliposome; oxaliplatin.

MeSH terms

  • Biomimetics
  • Colonic Neoplasms* / drug therapy
  • Colonic Neoplasms* / metabolism
  • Colorectal Neoplasms* / drug therapy
  • Colorectal Neoplasms* / pathology
  • Fluorouracil / therapeutic use
  • Humans
  • Hyperthermia, Induced*
  • Iron
  • Oxaliplatin / therapeutic use
  • Receptors, G-Protein-Coupled / metabolism

Substances

  • Fluorouracil
  • Oxaliplatin
  • Iron
  • Receptors, G-Protein-Coupled
  • LGR5 protein, human

Grants and funding

This research was funded by Project “PI19/01478-FEDER”, funded by Instituto de Salud Carlos III (ISCIII) and co-funded by the European Union, Junta de Andalucía (P20_00208), FEDER Operational Program (B-BIO-432-UGR20) and Project PDC2021-121135.100 funded through MCIN/AEI/10.13039/501100011033 and the European Union Next GenerationEU/ PRTR. This research was also funded in part by Personalized Medicine and Advanced Therapies Program (PMPTA22/00136), Instituto de Salud Carlos III.