rPIV5- and AVLP-vectored vaccine development with public tumor-specific neoantigens

摘要 本申请是为了回应被确认为非CA-CA的特殊利益通知(NOSI)而提交的 23-045.建议的补充计划旨在验证总共10个公共肿瘤的免疫原性- 我们的父母R01CA252713项目确定了特定的新抗原(PTSNA)候选基因，名为“犬MHC- I基因分型和肿瘤特异性新抗原测定“。它们被定义为突变多肽，它们是：1) 来源于犬和人类癌症的热点突变；以及2)被预测与MHC-I类(MHC-I)结合 在犬/人种群和/或亚种群(品种或种族)中占优势的等位基因。 我们将与两位领先的疫苗研发人员何彪和董安博士合作进行两项研究。1) 我们将把pTSNA候选基因串联克隆到疫苗载体副流感病毒5(PIV5)和PIV5- 基于自我放大的病毒样颗粒(AVLP)。由何博士和安博士开发的PIV5和AVLP是理想的 疫苗载体，特别是用于癌症疫苗开发的疫苗载体，因为它们可以诱导更强大的细胞免疫 回应，与其他策略相比。2)感染外周血单核细胞(PBMC) 从携带rPIV5-pTSNA和rAVLP-pTSNA病毒所需MHC-I等位基因的狗身上分离出，鉴定 通过质谱学分析MHC-I等位基因呈现的pTSNAs，鉴定免疫原性pTSNAs 通过干扰素-g检测。 这项拟议的补充研究将显著提高亲本R01的目标2c，通过加速 免疫原性pTSNA的发现。此外，这项研究将产生关键试剂(rPIV5-pTSNA和rAVLP- PTSNA疫苗)和未来影响大量人口的癌症疫苗开发的知识 无论是狗还是人类。考虑到基于PIV5的疫苗被证明是 对狗来说是安全的和免疫原性的，基于PIV5的疫苗通常是经鼻腔给药的。

Abstract This application is being submitted in response to the Notice of Special Interest (NOSI) identified as NOT-CA- 23-045. The proposed supplement project aims to validate the immunogenicity of a total 10 public tumor- specific neoantigen (pTSNA) candidates identified by our parent R01CA252713 project, entitled “Canine MHC- I genotyping and tumor specific neoantigen determination”. These are defined as mutant peptides that are: 1) derived from hotspot mutations in canine and human cancers; and 2) predicted to bind MHC class I (MHC-I) alleles dominant in canine/human populations and/or subpopulations (breeds or ethnic groups). We will collaborate with Drs. Biao He and Dong An, two leading vaccine developers to perform two studies. 1) We will clone the pTSNA candidates in tandem into vaccine vectors parainfluenza virus 5 (PIV5) and PIV5- based self-amplifying virus-like particle (AVLP). PIV5 and AVLP, developed by Drs He and An, are ideal vaccine vectors especially for cancer vaccine development, as they elicit more robust cellular immune responses, compared to other strategies. 2) We will infecting peripheral blood mononuclear cells (PBMCs) isolated from dogs harboring the required MHC-I alleles with rPIV5-pTSNA and rAVLP-pTSNA viruses, identify pTSNAs that are presented by the MHC-I alleles via mass-spectrometry, and identify immunogenic pTSNAs via interferon-g assays. This proposed supplement study will significantly enhance Aim 2c of the parent R01, by accelerating immunogenic pTSNA discovery. Furthermore, the study will yield critical reagents (rPIV5-pTSNA amd rAVLP- pTSNA vaccines) and knowledge for future cancer vaccine development that impacts a large population of both dogs and humans. The study is especially significant, considering that PIV5-based vaccines are shown to be safe and immunogenic for dogs, and PIV5-based vaccines are often administrated intranasally.