Cancer/testis antigens (CTAs), also known as tumor-specific antigens (TSAs) are specifically expressed in cancer cells and exhibit high immunogenicity, making them promising targets for immunotherapy and cancer vaccines. A new integrated high-throughput screening methodology for CTAs was proposed in this study through combining DNA methylation and RNA sequencing data. Briefly, the genes with increased transcript level and decreased DNA methylation were identified by multi-omics analysis. RNA sequencing studies in cell lines exposed to DNA methyltransferase (DNMT) inhibitors were performed to validate the inherent causal relationship between DNA hypomethylation and gene expression upregulation. We proposed a new integrated high-throughput screening methodology for identification of CTAs using multi-omics analysis. In addition, we tested the feasibility of this method using gastric cancer (GC) as an example. In GC, we identified over 2000 primary candidate CTAs and ultimately identified 20 CTAs with significant tissue-specificity, including a testis-specific serine protease TESSP1/PRSS41. Integrated analysis confirmed that PRSS41 expression was reactivated in gastrointestinal cancers by promoter DNA hypomethylation at the CpG site (cg08104780). Additionally, DNA hypomethylation of PRSS41 predicted a poor prognosis in GC. We propose a new high-throughput screening method for the identification of CTAs in cancer and validate its effectiveness. Our work emphasizes that serine protease PRSS41 is a novel TSA that is reactivated in GC due to promoter DNA hypomethylation.

中文翻译：

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利用多组学数据发现癌症睾丸抗原的新高通量筛选方法

癌症/睾丸抗原（CTA），也称为肿瘤特异性抗原（TSA），在癌细胞中特异性表达并表现出高免疫原性，使其成为免疫治疗和癌症疫苗的有希望的靶标。本研究通过结合 DNA 甲基化和 RNA 测序数据，提出了一种新的 CTA 集成高通量筛选方法。简而言之，通过多组学分析鉴定了转录水平增加和 DNA 甲基化减少的基因。对暴露于 DNA 甲基转移酶 (DNMT) 抑制剂的细胞系进行 RNA 测序研究，以验证 DNA 低甲基化与基因表达上调之间的内在因果关系。我们提出了一种新的集成高通量筛选方法，用于使用多组学分析来鉴定 CTA。此外，我们以胃癌（GC）为例测试了该方法的可行性。在 GC 中，我们鉴定了 2000 多个主要候选 CTA，并最终鉴定了 20 个具有显着组织特异性的 CTA，其中包括睾丸特异性丝氨酸蛋白酶 TESSP1/PRSS41。综合分析证实，胃肠道癌症中 PRSS41 的表达通过 CpG 位点 (cg08104780) 的启动子 DNA 低甲基化而重新激活。此外，PRSS41 的 DNA 低甲基化预示着 GC 的预后不良。我们提出了一种新的高通量筛选方法来鉴定癌症中的 CTA 并验证其有效性。我们的工作强调丝氨酸蛋白酶 PRSS41 是一种新型 TSA，由于启动子 DNA 低甲基化而在 GC 中重新激活。