FANCA 促进肺腺癌的进展，是表位疫苗免疫治疗的潜在靶点。
FANCA promotes lung adenocarcinoma progression and is a potential target for epitope vaccine immunotherapy.
机构信息
Shengli Clinical Medical College of Fujian Medical University, Fuzhou, China.
Department of Clinical Laboratory, Fuzhou University Affiliated Provincial Hospital, Fuzhou, China.
出版信息
J Transl Med. 2024 Oct 7;22(1):911. doi: 10.1186/s12967-024-05675-w.
BACKGROUND
FANCA mutations have been detected in a variety of cancers and found to be pro-carcinogenic. However, no functional studies have been identified regarding the involvement of FANCA in the occurrence and the immune response of LUAD.
METHODS
The mRNA expression and overall survival rates of FANCA were evaluated by the TIMER, PrognoScan and TCGA database in LUAD tissues, and FANCA expression was further validated by clinical serum samples using ELISA. The correlation between FANCA and immune infiltration level was investigated via TISIDB database and CIBERSORT algorithm. The Kaplan-Meier plotter was used to further evaluate the prognostic value based on the expression levels of FANCA in related immune cells. Then, the influence of FANCA knockout on the proliferation, migration, and invasion of A549 and H1299 cells was validated using CCK8, cloning formation, and Transwell assays. Subsequently, HLA-A2-restricted FANCA antigenic peptides were predicted and synthesized by NetMHC4.0 and SYFPEITHI, and DCs were induced and cultured in vitro. Finally, DCs loaded with HLA-A2-restricted FANCA antigenic peptides were co-cultured with autologous peripheral blood lymphocyte to generate specific CTLs. The killing effects of different CTLs on LUAD cells were studied.
RESULTS
The results showed that high levels of FANCA in patients with LUAD were significantly correlated with worse OS survival, which was correlated with age, clinical stage, pathological T stage, M stage, and N stage in LUAD. Knockdown of FANCA in A549 and H1299 cells significantly inhibited proliferation, metastasis, and invasion in vitro. In addition, FANCA was significantly related to immune infiltrate, genomic alterations and TMB. FANCA expression infuenced the prognosis of LUAD patients by directly affecting immune cell infltration. Finally, HLA-A2-restricted FANCA antigenic peptides were synthesized. And FANCA 146-154 (SLLEFAQYL) antigenic peptide exhibit a stronger affinity for DCs, and induce CTLs to produce stronger targeted killing ability for LUAD cells at an effector-to-target ratio of 40:1.
CONCLUSION
These results demonstrated that the elevation of FANCA promotes malignant phenotype of LUAD, and the potential peptide P2 (SLLEFAQYL) derived from FANCA may be used as an epitope vaccine for the treatment of LUAD.
背景
FANCA 突变已在多种癌症中被检测到，并被发现具有致癌作用。然而，目前尚未有关于 FANCA 参与 LUAD 的发生和免疫反应的功能研究。
方法
通过 TIMER、PrognoScan 和 TCGA 数据库评估 LUAD 组织中 FANCA 的 mRNA 表达和总生存率，并使用 ELISA 进一步验证临床血清样本中的 FANCA 表达。通过 TISIDB 数据库和 CIBERSORT 算法研究 FANCA 与免疫浸润水平的相关性。Kaplan-Meier plotter 用于根据相关免疫细胞中 FANCA 的表达水平进一步评估预后价值。然后，通过 CCK8、克隆形成和 Transwell 测定验证 FANCA 敲除对 A549 和 H1299 细胞增殖、迁移和侵袭的影响。随后，通过 NetMHC4.0 和 SYFPEITHI 预测和合成 HLA-A2 限制性 FANCA 抗原肽，并在体外诱导和培养 DC。最后，将负载 HLA-A2 限制性 FANCA 抗原肽的 DC 与自体外周血淋巴细胞共培养，产生特异性 CTL。研究了不同 CTL 对 LUAD 细胞的杀伤作用。
结果
结果表明，LUAD 患者中高水平的 FANCA 与更差的 OS 生存显著相关，与 LUAD 中的年龄、临床分期、病理 T 分期、M 分期和 N 分期相关。在 A549 和 H1299 细胞中敲低 FANCA 显著抑制体外增殖、转移和侵袭。此外，FANCA 与免疫浸润、基因组改变和 TMB 显著相关。FANCA 表达通过直接影响免疫细胞浸润影响 LUAD 患者的预后。最后，合成了 HLA-A2 限制性 FANCA 抗原肽。并且 FANCA 146-154（SLLEFAQYL）抗原肽对 DC 具有更强的亲和力，并在效应物与靶细胞比为 40:1 时诱导 CTL 对 LUAD 细胞产生更强的靶向杀伤能力。
结论
这些结果表明，FANCA 的升高促进了 LUAD 的恶性表型，并且源自 FANCA 的潜在肽 P2（SLLEFAQYL）可作为治疗 LUAD 的表位疫苗。
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