Yaseen Allah Rakha, Suleman Muhammad, Alzahrani Khalid J, Alsharif Khalaf F, Alzahrani Fuad M
School of Biological Sciences, Faculty of Life Sciences, University of the Punjab, Lahore, 54590, Pakistan.
Department of Clinical Laboratories Sciences, College of Applied Medical Sciences, Taif University, P.O. Box 11099, 21944, Aţ Ţā'if, Saudi Arabia.
Med Oncol. 2025 Jun 28;42(8):295. doi: 10.1007/s12032-025-02863-6.
Clear cell sarcoma (CCS) is a rare and aggressive soft tissue malignancy characterized by a high metastatic propensity and recurrence rate, with conventional non-specific treatment modalities often yielding limited efficacy and substantial adverse effects. This study aimed to design novel vaccine candidates: a multi-epitope vaccine (MEV) and an mRNA-based vaccine, both targeting the EWSR1-ATF1 and EWSR1-CREB1 fusion proteins, along with the PRAME antigen, to stimulate robust tumor-specific immune responses. Advanced immunoinformatics approaches were employed to identify highly antigenic B-cell and T-cell epitopes while minimizing potential allergenicity and toxicity, ensuring optimal safety and efficacy. The MEV construct was engineered with GM-CSF as an adjuvant to enhance dendritic cell activation and antigen presentation, with EAAK and AAY linkers incorporated to improve structural integrity and epitope processing. For the mRNA vaccine, the MEV was codon-optimized and incorporated into a stable mRNA construct with a 5' cap, Kozak sequence, and poly(A) tail to enhance the translation efficiency and prolong antigen expression. Structural and molecular dynamics simulations confirmed strong and stable interactions of both vaccine constructs with Toll-Like Receptor-3 (TLR-3), supporting their potential for effective immune activation. Furthermore, population coverage analysis demonstrated a global reach of 99.48%, ensuring broad immunogenicity across diverse genetic backgrounds. In silico immune response simulations predicted a sustained immune activation lasting over 417 days, characterized by robust cytokine secretion, strong memory cell formation, and high antibody titers following a three-dose regimen. These findings suggest that both the multi-epitope and mRNA-based vaccine candidates hold substantial promise as novel and precise immunotherapeutic interventions for CCS, potentially overcoming the limitations of existing treatment approaches and significantly enhancing long-term patient prognosis.
透明细胞肉瘤(CCS)是一种罕见且侵袭性强的软组织恶性肿瘤,其特点是具有高转移倾向和复发率,传统的非特异性治疗方式往往疗效有限且不良反应严重。本研究旨在设计新型候选疫苗:一种多表位疫苗(MEV)和一种基于mRNA的疫苗,二者均靶向EWSR1-ATF1和EWSR1-CREB1融合蛋白以及PRAME抗原,以刺激强大的肿瘤特异性免疫反应。采用先进的免疫信息学方法来识别高抗原性的B细胞和T细胞表位,同时将潜在的致敏性和毒性降至最低,确保最佳的安全性和有效性。MEV构建体经工程改造,以GM-CSF作为佐剂来增强树突状细胞的激活和抗原呈递,并掺入EAAK和AAY接头以改善结构完整性和表位加工。对于mRNA疫苗,对MEV进行密码子优化,并将其整合到具有5'帽、Kozak序列和聚腺苷酸尾的稳定mRNA构建体中,以提高翻译效率并延长抗原表达。结构和分子动力学模拟证实了两种疫苗构建体与Toll样受体-3(TLR-3)之间有强大且稳定的相互作用,支持了它们有效激活免疫的潜力。此外,群体覆盖率分析表明全球覆盖率达99.48%,确保了在不同遗传背景下具有广泛的免疫原性。计算机免疫反应模拟预测,在三剂接种方案后,持续的免疫激活将持续超过417天,其特征为强大的细胞因子分泌、强烈的记忆细胞形成和高抗体滴度。这些发现表明,多表位疫苗和基于mRNA的候选疫苗作为CCS的新型精准免疫治疗干预措施都具有巨大潜力,有可能克服现有治疗方法的局限性,并显著改善患者的长期预后。
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