pH-Dependent Release and Folate Receptor Alpha (FRα) Targeting To Allow Exclusive Targeting of Lipid-Nanoparticles (LNP)

Abstract

Introduction: Colorectal cancer rates continue to rise in Canada, placing an increasing economic burden on the healthcare system and profoundly impacting patients’ quality of life. Here, we propose a novel method for delivering anti-colorectal cancer treatment that uses a Eudragit-coated folate-LNP conjugation to allow pH-sensitive release and specificity for cancerous colon cells.

Methods: GFP-tagged IL-10 mRNA will serve as a biomarker to evaluate folate receptor alpha (FR  binding, transfection, and release of therapeutic contents by the novel LNP formulation in LS174 human cells, measured by flow cytometry. To confirm exclusive delivery, tissue samples from the small and large intestines of Apc^Min/+ mice treated with the formulation will be analyzed using ELISA following oral gavage. MTT assay of healthy, non-cancerous CCD 841 CoN will confirm consistent cell viability across non-targeted colonic tissue.

Expected Results: The folate-LNP formulation is expected to bind FR  on the LS174 cells efficiently and release the encapsulated IL-10 mRNA after endosomal escape, evidenced by significantly higher fluorescence in treated cells than the negative controls. In the mouse model, the Eudragit coating is anticipated to dissolve exclusively in the colonic tissue of the mouse models, with minimal dissolution in the small intestine, allowing targeted binding to cancerous colonic enterocytes.

Discussion: Our proposed folate-LNP formulation is designed to achieve targeted delivery to colonic tissue by leveraging pH-sensitive release and FR  upregulation in colorectal cancer cells. Efficient binding and successful translation of IL-10 mRNA in the LS174 cells will demonstrate the formulation’s specificity and therapeutic potential. Exclusive localization to cancerous colonic tissue in the mouse model will confirm the ability to bypass non-target tissues.

Conclusion: This approach represents the first known proposal for a Eudragit-coated folate-LNP molecule targeting the colon. If successful, this approach could offer a more effective, targeted treatment for colorectal cancer, minimizing systemic side effects and improving patient outcomes. Future research and clinical validation are required to confirm the safety and efficacy of this approach, as well as to determine the patients who will benefit most.