Microfluidic Synthesis of miR-10a Antisense Oligonucleotides-Loaded LNP Combined with Cisplatin for Colorectal Cancer Therapy.

Abstract

Introduction: To address the issue of insufficient sensitivity of the chemotherapeutic drug cisplatin in colorectal cancer (CRC), this study aimed to develop a miR-10a antisense oligonucleotide (AMO) system delivered by lipid nanoparticles (LNPs). The aim was to enhance the sensitivity and efficacy of cisplatin by targeting and inhibiting the expression of miR-10a in CRC.

Methods: Through bioinformatics analysis, key miRNAs that are differentially expressed in colon cancer were identified. Then, by using the FDA-approved patisiran formulation and microfluidic technology, LNPs/miR10a- AMOs were constructed to target miR-10a. The transfection efficiency, anti-tumor effectiveness, and safety were evaluated through in vivo and in vitro experiments.

Results: LNPs increased the transfection efficiency of AMOs by 2.3 times (p < 0.01). In HCT116 cells, miR- 10a expression was inhibited by 68.5%, and after combined treatment with cisplatin, the IC50 decreased from 8.7 μM to 5.0 μM (p < 0.001). Meanwhile, cell viability in normal THLE2 and HCoEpiC cells remained > 85%. In the xenograft model, the combined therapy achieved a 100.4% tumor growth inhibition compared to cisplatin monotherapy, with Ki67 expression decreased by 67.3%, and cleaved-caspase-3 expression increased by 3.1- fold (p<0.001). Histopathological analysis confirmed minimal organ toxicity. Mechanistically, miR-10a downregulation sensitized the tumor by activating the apoptotic pathway and inhibiting proliferation.

Discussion: Our findings demonstrated that targeting miR-10a significantly enhanced the sensitivity of tumor cells to cisplatin, providing a novel strategic approach to overcome chemoresistance. Compared to existing studies, the innovation of this work lies in employing an FDA-approved LNP formulation, substantially improving the feasibility for clinical translation. Furthermore, the application of microfluidic technology ensured reproducible nanoparticle preparation, offering distinct advantages over conventional methods in terms of industrial applicability. However, this study has involved certain limitations; the animal models utilized could not fully recapitulate the complexity of the human tumor microenvironment, and the precise molecular mechanisms regulated by miR-10a warrant further investigation.

Conclusion: LNPs/miR-10a-AMOs can efficiently deliver AMOs and represent a potential strategy for treating CRC. By enhancing the sensitivity of tumors to cisplatin, this system improved the therapeutic effect while ensuring the safety of normal cells. This provides a new strategy with translational potential for the precise treatment of CRC.