肝细胞癌mRNA疫苗的肿瘤相关抗原筛选及验证-现代医学

肝细胞癌mRNA疫苗的肿瘤相关抗原筛选及验证

单位：1. 南京医科大学公共卫生学院, 江苏南京 211166;
2. 南京市第二医院传染病科, 江苏南京 211103;
3. 南京市第二医院南京医科大学公共卫生学院联合培养基地, 江苏南京 210008

分类号：R735.7

出版年·卷·期（页码）：2026·45·第四期（597-607）

摘要：

目的： 通过生物信息学数据筛选能利用于肝细胞癌(HCC) mRNA疫苗的肿瘤相关抗原(TAAs)。方法： 整合TCGA、cBioPortal数据库分析HCC基因突变与异常表达情况；利用GEPIA 2.0和Kaplan-Meier评估基因过表达对预后的影响；通过TIMER分析基因与抗原递呈细胞(APCs)免疫浸润的相关性；采用TSNAdb预测抗原免疫原性；结合临床转录组数据及人类蛋白质图谱数据库(HPA)进行表达验证与免疫组化分析。结果： 对TCGA-LIHC的数据初步筛选获得1 485个与HCC相关的过表达基因；生存分析发现108个与患者总生存期(OS)和无复发生存期(RFS)相关的基因。两者取交集得到33个潜在TAAs；进一步分析其与APCs免疫浸润的关联性、与HLA Ⅰ类分子的结合能力，确定核转运蛋白α2亚基(KPNA2)、细胞周期蛋白B1(CCNB1)、细胞周期蛋白依赖性激酶1(CDK1)、核糖体生物合成调控因子1(PES1)、极光激酶A(AURKA)和检查点激酶1(CHEK1)6个HCC-TAAs可作为HCC mRNA疫苗的候选靶点；并在免疫组化结果和小样本的临床转录组测序分析中得到较好的一致性验证。结论： 通过多维生物信息学数据能够筛选具有潜力的HCC-TAAs，为后续mRNA疫苗的开发和临床转化应用提供参考依据。

Objective: To screen tumor-associated antigens(TAAs) applicable to mRNA vaccines for hepatocellular carcinoma(HCC) using bioinformatics data. Methods: Gene mutations and abnormal expression in HCC were analyzed by integrating TCGA and cBioPortal databases. The impact of gene overexpression on prognosis was evaluated using GEPIA 2.0 and Kaplan-Meier analysis. The correlation between gene expression and immune infiltration of antigen-presenting cell(APC) was analyzed via TIMER. Antigen immunogenicity was predicted using TSNAdb. Expression validation and immunohistochemical analysis were performed using clinical transcriptomic data and the Human Protein Atlas(HPA). Results: Preliminary screening of TCGA-LIHC data identified 1 485 genes overexpressed in HCC. Survival analysis revealed 108 genes associated with both overall survival(OS) and recurrence-free survival(RFS) of patients. Intersection of these two datasets yielded 33 potential TAAs. Further analysis of their immune infiltration correlation with APCs and their binding affinity to HLA class Ⅰ molecules identified six HCC-TAAs as candidate targets for HCC mRNA vaccines: karyopherin subunit alpha 2(KPNA2), cyclin B1(CCNB1), cyclin-dependent kinase 1(CDK1), pescadillo ribosomal biogenesis factor 1(PES1), aurora kinase A(AURKA), and checkpoint kinase 1(CHEK1). These findings were consistently validated by immunohistochemistry and transcriptomic data. Conclusion: Multi-dimensional bioinformatics data enable the screening of potential HCC-TAAs, providing a reference basis for the development of mRNA vaccines and their clinical translation.

参考文献：

[1] 郑荣寿,陈茹,韩冰峰,等.2022年中国恶性肿瘤流行情况分析[J].中华肿瘤杂志,2024,46(03)：221-231.
[2] BIZUAYEHU H M,AHMED K Y,KIBRET G D,et al.Global disparities of cancer and its projected burden in 2050[J].JAMA Netw Open,2024,7(11):e2443198.
[3] FORNER A,REIG M,BRUIX J.Hepatocellular carcinoma[J].Lancet,2018,391(10127):1301-1314.
[4] MALUCCIO M,COVEY A.Recent progress in understanding,diagnosing,and treating hepatocellular carcinoma[J].CA Cancer J Clin,2012,62(6):394-399.
[5] ANWANWAN D,SINGH S K,SINGH S,et al.Challenges in liver cancer and possible treatment approaches[J].Biochim Biophys Acta Rev Cancer,2020,1873(1):188314.
[6] SU G L,ALTAYAR O,O'SHEA R,et al.AGA clinical practice guideline on systemic therapy for hepatocellular carcinoma[J].Gastroenterology,2022,162(3):920-934.
[7] MCNAMARA M A,NAIR S K,HOLL E K.RNA-based vaccines in cancer immunotherapy[J].J Immunol Res,2015,2015:794528.
[8] LORENTZEN C L,HAANEN J B,MET Ö,et al.Clinical advances and ongoing trials on mRNA vaccines for cancer treatment[J].Lancet Oncol,2022,23(10):e450-e458.
[9] YUAN Y,GAO F,CHANG Y,et al.Advances of mRNA vaccine in tumor:a maze of opportunities and challenges[J].Biomark Res,2023,11(1):6.
[10] ZHANG T Y,XU H,ZHENG X N,et al.Clinical benefit and safety associated with mRNA vaccines for advanced solid tumors:a meta-analysis[J].MedComm,2023,4(4):e286.
[11] REN Y,YUE Y,LI X,et al.Proteogenomics offers a novel avenue in neoantigen identification for cancer immunotherapy[J].Int Immunopharmacol,2024,142(Pt A):113147.
[12] SCHUMACHER T N,SCHREIBER R D.Neoantigens in cancer immunotherapy[J].Science,2015,348(6230):69-74.
[13] GREES M,SHARBI-YUNGER A,EVANGELOU C,et al.Optimized dendritic cell vaccination induces potent CD8 T cell responses and anti-tumor effects in transgenic mouse melanoma models[J].Oncoimmunology,2018,7(7):e1445457.
[14] 高花,韩勇,翟晓鑫,等.MHC分子抗原递呈机制的研究进展[J].生命科学,2017,29(5):450-461.
[15] LI T,FU J,ZENG Z,et al.TIMER2.0 for analysis of tumor-infiltrating immune cells[J].Nucleic Acids Res,2020,48(W1):W509-W514.
[16] GU W,LI G,ZHANG W,et al.miR-29b regulates Th2 cell differentiation in asthma by targeting inducible B7-H3 and STAT3[J].Clin Transl Allergy,2022,12(1):e12114.
[17] NEIZER-ASHUN F,BHATTACHARYA R.Reality CHEK:understanding the biology and clinical potential of CHK1[J].Cancer Lett,2021,497:202-211.
[18] SMITS V A J,GILLESPIE D A.DNA damage control:regulation and functions of checkpoint kinase 1[J].FEBS J,2015,282(19):3681-3692.
[19] SUN J,ZHU Z,LI W,et al.UBE2T-regulated H2AX monoubiquitination induces hepatocellular carcinoma radioresistance by facilitating CHK1 activation[J].J Exp Clin Cancer Res,2020,39(1):222.
[20] YIN C,SUN Y,LI H,et al.miR-424-5p suppresses tumor growth and progression by directly targeting CHEK1 and activating cell cycle pathway in Hepatocellular Carcinoma[J].Heliyon,2024,10(18):e37769.
[21] WANG Q,BODE A M,ZHANG T.Targeting CDK1 in cancer:mechanisms and implications[J].NPJ Precis Oncol,2023,7(1):58.
[22] WU C X,WANG X Q,CHOK S H,et al.Blocking CDK1/PDK1/β-Catenin signaling by CDK1 inhibitor RO3306 increased the efficacy of sorafenib treatment by targeting cancer stem cells in a preclinical model of hepatocellular carcinoma[J].Theranostics,2018,8(14):3737-3750.
[23] HU B,SHEN X,QIN W,et al.A prognostic nomogram for hepatocellular carcinoma based on wound healing and immune checkpoint genes[J].J Clin Transl Hepatol,2022,10(5):891-900.
[24] FANG Y,YU H,LIANG X,et al.Chk1-induced CCNB1 overexpression promotes cell proliferation and tumor growth in human colorectal cancer[J].Cancer Biol Ther,2014,15(9):1268-1279.
[25] XIE B,WANG S,JIANG N,et al.Cyclin B1/CDK1-regulated mitochondrial bioenergetics in cell cycle progression and tumor resistance[J].Cancer Lett,2019,443:56-66.
[26] GU J,LIU X,LI J,et al.microRNA-144 inhibits cell proliferation,migration and invasion in human hepatocellular carcinoma by targeting CCNB1[J].Cancer Cell Int,2019,19:15.
[27] LIN Z Z,HSU H C,HSU C H,et al.The Aurora kinase inhibitor VE-465 has anticancer effects in pre-clinical studies of human hepatocellular carcinoma[J].J Hepatol,2009,50(3):518-527.
[28] ZHU Q,YU X,ZHOU Z W,et al.Inhibition of aurora a kinase by alisertib induces autophagy and cell cycle arrest and increases chemosensitivity in human hepatocellular carcinoma HepG2 cells[J].Curr Cancer Drug Targets,2017,17(4):386-401.
[29] FU Z,JIAO Y,LI Y Q,et al.PES1 in liver cancer:a prognostic biomarker with tumorigenic roles[J].Cancer Manag Res,2019,11:9641-9653.
[30] FAN P,WANG B,MENG Z,et al.PES1 is transcriptionally regulated by BRD4 and promotes cell proliferation and glycolysis in hepatocellular carcinoma[J].Int J Biochem Cell Biol,2018,104:1-8.
[31] WANG J,SUN J,ZHANG N,et al.PES1 enhances proliferation and tumorigenesis in hepatocellular carcinoma via the PI3K/AKT pathway[J].Life Sci,2019,219:182-189.
[32] WEI S,LIU K,HE Q,et al.PES1 is regulated by CD44 in liver cancer stem cells via miR-105-5p[J].FEBS Lett,2019,593(14):1777-1786.
[33] HAN Y,WANG X.The emerging roles of KPNA2 in cancer[J].Life Sci,2020,241:117140.
[34] 张文福,李学东.KPNA2在肿瘤发生发展中作用的进展[J].现代医学,2016,44(05):759-762.
[35] GARRITANO S,INGA A,GEMIGNANI F,et al.More targets,more pathways and more clues for mutant p53[J].Oncogenesis,2013,2(7):e54.
[36] WINKLER J,ORI A,HOLZER K,et al.Prosurvival function of the cellular apoptosis susceptibility/importin-α1 transport cycle is repressed by p53 in liver cancer[J].Hepatology,2014,60(3):884-895.
[37] GUO X,WANG Z,ZHANG J,et al.Upregulated KPNA2 promotes hepatocellular carcinoma progression and indicates prognostic significance across human cancer types[J].Acta Biochim Biophys Sin,2019,51(3):285-292.
[38] JIANG P,TANG Y,HE L,et al.Aberrant expression of nuclear KPNA2 is correlated with early recurrence and poor prognosis in patients with small hepatocellular carcinoma after hepatectomy[J].Med Oncol,2014,31(8):131.
[39] YANG Y,GUO J,HAO Y,et al.Silencing of karyopherin α2 inhibits cell growth and survival in human hepatocellular carcinoma[J].Oncotarget,2017,8(22):36289-36304.
[40] LI Y F,HOU Q Q,ZHAO S,et al.Identification of tumor-specific neoantigens and immune clusters of hepatocellular carcinoma for mRNA vaccine development[J].J Cancer Res Clin Oncol,2023,149(2):623-637.
[41] LIAO W,SHI Z,TANG H,et al.Identifying potential tumor antigens and antigens-related subtypes in hepatocellular carcinoma for mRNA vaccine development[J].J Oncol,2022,2022:6851026.

服务与反馈：

【文章下载】【发表评论】【查看评论】【加入收藏】

提示：您还未登录，请登录！点此登录