Innovative Dual mRNA-Lipid Nanoparticle Therapy Targeting CRHBP and CFHR3 for Enhanced Treatment of Hepatocellular Carcinoma

Tianmei Fu,1,&ast; Boxuan Zhou,2,&ast; Yingliang Li,2,&ast; Wei Liu,1 Yuankang Xie,3 Zhaohong Mo,4 Fang Yin,1 Yu Wang,1 Kang Fang,5 Yangyang Fang,1 Ziqing Xiong,1 Kuai Yu,1 Aiping Le1 1Department of Transfusion Medicine, Key Laboratory of Jiangxi Province for Transfusion Medicine, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, People’s Republic of China; 2Department of Breast Disease Center, the First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, People’s Republic of China; 3Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Gannan Medical University, Ganzhou, People’s Republic of China; 4Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital of Guangxi Medical University, Nanchang, People’s Republic of China; 5Department of Hepatobiliary and Pancreatic Surgery, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Kuai Yu; Aiping Le, Email [email protected]; [email protected]: Hepatocellular carcinoma (HCC) is a deadly disease requiring the identification of new therapeutic targets and strategies.Methods: This study identified genes linked to HCC progression via differential analysis. Key genes were identified through univariate and multivariate Cox regression analysis. The biological effects of co-expressed CRHBP and CFHR3 were evaluated in vitro. mRNAs encoding CRHBP and CFHR3 were encapsulated in lipid nanoparticles (LNPs), with the addition of SP94 peptide on the LNPs surface to enhance targeting. The therapeutic efficacy of dual-mRNA LNPs was evaluated in HCC cells and mouse models.Results: CRHBP and CFHR3 were closely associated with HCC progression. Low expression of CRHBP (P < 0.01, HR = 1.931 [1.174– 3.175]) and CFHR3 (P < 0.05, HR = 1.755 [1.066– 2.890]) was identified as a poor prognostic factor for HCC. The risk score model combining CRHBP and CFHR3 demonstrated superior predictive power (P < 0.001, HR = 2.935 [1.768– 4.872]). Co-expression of CRHBP and CFHR3 significantly inhibited the malignant biological functions of HCC cells. Treatment with SP94 peptide-modified dual-mRNA LNPs markedly suppressed HCC tumor growth and exhibited excellent biocompatibility and safety.Conclusion: Our study proposes a dual-targeted therapeutic strategy for HCC, which may represent a promising treatment approach. Keywords: CRHBP, CFHR3, Lipid nanoparticles, mRNA