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

摘要 本申请是为了响应被标识为NOT-CA的特别利益通知（NOSI）而提交的- 23-045.该补充项目旨在验证10种公共肿瘤的免疫原性， 特异性新抗原（pTSNA）候选物，其由我们的父R 01 CA 252713项目，题为“犬MHC- I基因分型和肿瘤特异性新抗原测定”。这些被定义为突变肽，其为：1） 来源于犬和人癌症中的热点突变;和2）预测结合MHC I类（MHC-I） 等位基因在犬/人类群体和/或亚群（品种或种族群体）中占优势。 我们将与两位领先的疫苗开发者何彪和董安博士合作进行两项研究。第一章 我们将pTSNA候选物串联克隆到疫苗载体副流感病毒5型（PIV 5）和PIV 5 - 1中。 自扩增病毒样颗粒（AVLP）。由何博士和安博士开发的PIV 5和AVLP是理想的 疫苗载体，特别是用于癌症疫苗开发，因为它们引起更强大的细胞免疫 与其他策略相比。2)我们将感染外周血单个核细胞（PBMC） 从携带所需MHC-I等位基因的犬中分离rPIV 5-pTSNA和rAVLP-pTSNA病毒，鉴定 通过质谱分析由MHC-I等位基因呈递的pTSNA，并鉴定免疫原性pTSNA 通过干扰素-G测定。 这项拟议的补充研究将通过加速 免疫原性pTSNA发现。此外，该研究将产生关键试剂（rPIV 5-pTSNA和rAVLP-pTSNA）。 pTSNA疫苗）和未来癌症疫苗开发的知识， 狗和人类都是。这项研究尤其重要，因为基于PIV 5的疫苗显示出 对于狗是安全的和免疫原性的，并且基于PIV 5的疫苗通常鼻内施用。

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.