Toward translation of a plant virus-based in situ vaccination nanotechnology

总结 该R 01提交给了纳米技术癌症干预（TTNCI）项目的翻译， 是我们成功的U 01 CA 218292（09/2017-07/2022）的延续，通过NCI联盟资助， 纳米技术在癌症计划。在U 01项目期间，我们证明了我们的纳米颗粒 来源于植物病毒豇豆花叶病毒（CPMV）刺激有效的抗肿瘤免疫反应， 多种肿瘤小鼠模型。此外，在患有黑色素瘤、肉瘤和乳腺癌的伴侣犬中进行的试验 通过我们独特的免疫治疗方法证明了强大的抗肿瘤功效。CPMV是一种肿瘤内 注射原位疫苗（ISV）技术，能够诱导持久的免疫介导的抗肿瘤 对治疗和未治疗的肿瘤的功效以及对复发的免疫记忆。 从机制上讲，CPMV激活先天免疫系统以改善肿瘤相关的和非肿瘤性免疫系统的处理。 新抗原，从而导致针对由以下抗原表达的抗原的功能性更强的适应性抗肿瘤免疫： 肿瘤这导致治疗和未治疗的恶性肿瘤的缓解，并防止复发 通过自适应手臂和免疫记忆。在数据和知识的强大基础上， 该项目将专注于优化GMP生产和小鼠肿瘤模型的药理学 和犬类患者，为未来的人类肿瘤学试验提供基础。多PI领导 团队斯坦梅茨（加州大学圣地亚哥分校），Fiering（达特茅斯）和Ranjan（俄勒冈州立大学）带来了免费的专业知识， 基于植物病毒的CPMV纳米技术、癌症免疫学和兽医肿瘤学/免疫治疗试验。 我们的团队还得到了一组具有监管专业知识的顾问（Garnick，Mosaic IE Inc.）的支持， 药理学/毒理学（Luksic，Intrinsik）、临床（Garoadine，Mosaic IE Inc.）事务，以及经验 垂直农业（Eisenberg，OPO）。我们相信，我们强大的合作者团队可以帮助建立 未来的植物源生物制品。我们将实现以下具体目标：（1）化学， 用于CPMV的规模化生产和QAC的生产和控制（CMC）方法开发- ISV，（2）药理学：确定CPMV-ISV候选药物在体内的生物分布、给药和毒理学。 原位和转移性黑色素瘤小鼠模型;和（3）犬试验：建立在黑色素瘤中的给药和安全性。 在适合原位接种的不同肿瘤类型（黑色素瘤、肉瘤和肥大细胞瘤）中进行的犬肿瘤学试验， 细胞瘤）。通过在小鼠肿瘤模型和狗患者中的平行试验，我们将确定活检或血液 用于纵向监测治疗反应的生物标志物。成功的完成将使我们处于有利地位 生产规范（GMP）生产这种植物病毒为基础的生物纳米技术，并继续 发展路径，例如通过NCI实验治疗（NExT）计划，从而将 基于CPMV的候选药物进入人体临床试验和实践。

Summary This R01, submitted to the Toward Translation of Nanotechnology Cancer Interventions (TTNCI) program, is a continuation of our successful U01 CA218292 (09/2017-07/2022), funded through the NCI Alliance for Nanotechnology in Cancer program. During the U01 project period, we demonstrated that our nanoparticles derived from the plant virus cowpea mosaic virus (CPMV) stimulate a potent antitumor immune response against multiple tumor mouse models. Additionally, trials in companion dogs with melanoma, sarcoma, and breast cancer demonstrate potent antitumor efficacy with our unique immunotherapy approach. CPMV is an intratumoral injected in situ vaccine (ISV) technology, with an ability to induce durable immune-mediated antitumor efficacy against treated and untreated tumors as well as immunological memory against recurrence. Mechanistically, CPMV activates the innate immune system to improve processing of tumor-associated and neoantigens, thereby resulting in a more functional adaptive antitumor immunity against antigens expressed by the tumor. This results in remissions of treated and untreated malignant tumors, and protection from recurrence via the adaptive arm and immune memory. Building on this strong foundation of data as well as intellectual property, this project will focus on optimizing GMP manufacturing and pharmacology in murine tumor models and canine patients to provide the foundational basis of future human oncology trials. The multi-PI leadership team Steinmetz (UC San Diego), Fiering (Dartmouth) and Ranjan (OSU) brings complimentary expertise in plant virus-based CPMV nanotechnology, cancer immunology, and veterinary oncology/immunotherapy trials. Our team is also supported by a group of consultants with expertise in regulatory (Garnick, Mosaic IE Inc.), pharmacology/toxicology (Luksic, Intrinsik), clinical (Garovoy, Mosaic IE Inc.) affairs, as well as experience in vertical farming (Eisenberg, OPO). We believe that our strong team of collaborators can help establish the future manufacture of the plant-derived biologic. We will fulfil the following Specific Aims: (1) Chemistry, Manufacturing, and Control (CMC) method development for scalable manufacture and QAC of the CPMV- ISV, (2) Pharmacology: Determine biodistribution, dosing, and toxicology of the CPMV-ISV drug candidate in orthotopic and metastatic mouse models of melanoma; and (3) Canine trials: establish dosing and safety in canine oncology trials in varying tumor types amenable for in situ vaccination (melanoma, sarcoma, and mast- cell tumor). Through parallel trials in murine tumor models and dog patients, we will identify biopsy or blood biomarkers for longitudinal monitoring of treatment response. Successful completion would position us for good manufacturing practice (GMP) production of this plant virus-based biologic nanotechnology and to continue the developmental path, e.g. through the NCI Experimental Therapeutics (NExT) program, thereby translating the CPMV-based drug candidate into a human clinical trial and practice.