反向疫苗学方法构建非小细胞肺癌表达的癌-睾丸抗原多表位疫苗

Reverse Vaccinology Approach in Constructing a Multi-Epitope Vaccine Against Cancer-Testis Antigens Expressed in Non-Small Cell Lung Cancer.

机构信息

Department of Physical Sciences, College of Science, Polytechnic University of the Philippines, Manila City, Philippines.

出版信息

Asian Pac J Cancer Prev. 2021 May 1;22(5):1495-1506. doi: 10.31557/APJCP.2021.22.5.1495.

BACKGROUND

The 5-year survival rate of non-small cell lung cancer (NSCLC) patients has not significantly improved despite advancements in the currently applied treatments. Thus, efforts are put forth in developing novel immunotherapeutic agents targeting cancer-testis antigens (CTA) in NSCLC. This work utilized reverse vaccinology approach in designing a novel multi-epitope vaccine targeting melanoma-associated antigen 3 (MAGEA3), MAGEA4, New York esophageal squamous cell carcinoma-1 (NY-ESO-1), and Kita-Kyushu lung cancer antigen 1 (KK-LC1), being the most frequently expressed CTAs in NSCLC.

METHODS

Epitopes were mapped from the sequences of CTAs. The population coverage (PC) of identified CD4+ and CD8+ epitopes were estimated. Candidate linear B cell (BL), CD4+, and CD8+ epitopes were adjoined in a multi-epitope construct (Mvax) with flagellin domain as an adjuvant. Antigenicity, and cross-reactivity of Mvax were examined. The tertiary structure of Mvax was modelled, and validated. All epitopes included in the vaccine were docked with their human leukocyte antigen (HLA) binders. The immunogenicity of epitopes in Mvax was validated through molecular dynamics analysis.

RESULTS

Mvax contains 22 epitopes from MAGEA3, MAGEA4, NY-ESO-1, and KK-LC1. It is classified as antigenic, non-allergen, non-toxic, and possesses physicochemical stability. Epitopes have no significant hits with other human proteins, except for 2 other CTAs frequently expressed in NSCLC. The stretch of BL epitopes in Mvax confers flexibility, and accessibility emphasizing its antigenicity. The tertiary structure analysis showed that Mvax model has good structural quality. All epitopes included in the vaccine are highly immunogenic as indicated by favorable binding affinity, low binding energy, and acceptable root-mean-square deviation (RMSD). CD4+ and CD8+ epitopes have global PC of 81.81%, and 84.15%, respectively.

CONCLUSION

Overall, in silico evaluations show that Mvax is a potential immunotherapeutic agent against NSCLC.

背景

尽管目前应用的治疗方法有所进步，但非小细胞肺癌（NSCLC）患者的 5 年生存率并没有显著提高。因此，人们正在努力开发针对癌症睾丸抗原（CTA）的新型免疫治疗药物。本研究利用反向疫苗学方法设计了一种针对黑色素瘤相关抗原 3（MAGEA3）、MAGEA4、纽约食管鳞状细胞癌-1（NY-ESO-1）和北九州肺癌抗原 1（KK-LC1）的新型多表位疫苗，这些抗原是 NSCLC 中最常表达的 CTA。

方法

从 CTA 的序列中映射出表位。估计鉴定的 CD4+和 CD8+表位的人群覆盖率（PC）。候选线性 B 细胞（BL）、CD4+和 CD8+表位与鞭毛蛋白结构域作为佐剂连接在一个多表位构建体（Mvax）中。检测 Mvax 的抗原性和交叉反应性。对 Mvax 的三级结构进行建模和验证。疫苗中包含的所有表位都与它们的人类白细胞抗原（HLA）结合物对接。通过分子动力学分析验证表位在 Mvax 中的免疫原性。

结果

Mvax 包含来自 MAGEA3、MAGEA4、NY-ESO-1 和 KK-LC1 的 22 个表位。它被归类为抗原性、非变应原性、非毒性，并且具有物理化学稳定性。除了 NSCLC 中经常表达的另外 2 个 CTA 外，这些表位与其他人类蛋白没有明显的相似性。Mvax 中的 BL 表位序列赋予其柔韧性和可及性，强调其抗原性。三级结构分析表明，Mvax 模型具有良好的结构质量。疫苗中包含的所有表位都具有高度的免疫原性，表现为良好的结合亲和力、低结合能和可接受的均方根偏差（RMSD）。CD4+和 CD8+表位的全球 PC 分别为 81.81%和 84.15%。