Nafiah Zahrotun, Ujiantari Navista Sri Octa, Rattyananda Badra Sanditya, Utami Setyowati Triastuti, Alwi Ratna Surya, Hermawan Adam
Laboratory of Macromolecular Engineering, Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Universitas Gadjah Mada Sekip Utara II, 55281, Yogyakarta, Indonesia.
Laboratory of Medicinal Chemistry, Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Universitas Gadjah Mada, Sekip Utara II, 55281, Yogyakarta, Indonesia.
Comput Biol Med. 2025 Sep;196(Pt B):110757. doi: 10.1016/j.compbiomed.2025.110757. Epub 2025 Jul 18.
A-Kinase Anchoring Protein 3 (AKAP3), a Cancer-Testis Antigen (CTA), is involved in cell proliferation and is aberrantly expressed in triple-negative breast cancer (TNBC), making it a promising target for immunotherapy. This study aimed to design a novel multi-epitope vaccine targeting AKAP3 using immunoinformatics approaches. Predicted CTL, HTL, and B-cell epitopes were selected based on their immunogenicity, antigenicity, non-allergenicity, and non-toxicity. The final construct integrated three epitopes from each category, connected by appropriate linkers (EAAAK, AAY, GPGPG, KK) and adjuvanted with the 50S ribosomal protein L7/L12 to enhance the immune response. Population coverage analysis indicated high accessibility, with 99.28 % in Indonesia, 99.4 % in Southeast Asia, and 100 % globally. Structural modeling and validation confirmed the vaccine's stability and immunogenic profile. Molecular docking demonstrated strong binding affinity to TLR-4 (-839.8 kcal/mol), supported by stable interactions in molecular dynamics simulations. Immune simulations further predicted the robust activation of both cellular and humoral immunity. In silico cloning confirmed compatibility with the pET28a(+) expression system for potential recombinant production. Collectively, these findings support the designed vaccine as a promising candidate for TNBC immunotherapy, warranting further in vitro and in vivo evaluation.
A激酶锚定蛋白3（AKAP3）是一种癌-睾丸抗原（CTA），参与细胞增殖，在三阴性乳腺癌（TNBC）中异常表达，使其成为免疫治疗的一个有前景的靶点。本研究旨在利用免疫信息学方法设计一种靶向AKAP3的新型多表位疫苗。根据其免疫原性、抗原性、非致敏性和无毒性选择预测的细胞毒性T淋巴细胞（CTL）、辅助性T淋巴细胞（HTL）和B细胞表位。最终构建体整合了来自每个类别的三个表位，通过适当的接头（EAAAK、AAY、GPGPG、KK）连接，并与50S核糖体蛋白L7/L12佐剂结合以增强免疫反应。群体覆盖率分析表明其具有高可及性，在印度尼西亚为99.28%，在东南亚为99.4%，在全球为100%。结构建模和验证证实了疫苗的稳定性和免疫原性特征。分子对接显示与Toll样受体4（TLR-4）具有强结合亲和力（-839.8千卡/摩尔），分子动力学模拟中的稳定相互作用支持了这一点。免疫模拟进一步预测了细胞免疫和体液免疫的强劲激活。电子克隆证实了与pET28a(+)表达系统的兼容性，以便进行潜在的重组生产。总体而言，这些发现支持所设计的疫苗作为TNBC免疫治疗的一个有前景的候选物，值得进一步进行体外和体内评估。
