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

摘要 本R01，提交给纳米技术癌症干预措施的转译(TTNCI)计划， 是我们成功的U01 CA218292(09/2017-07/2022)的延续，由NCI联盟资助 癌症中的纳米技术计划。在U01项目期间，我们展示了我们的纳米颗粒 从植物病毒中提取的豌豆花叶病毒(CPMV)能激发抗肿瘤免疫反应 多瘤小鼠模型。此外，在患有黑色素瘤、肉瘤和乳腺癌的伴侣犬身上进行的试验 通过我们独特的免疫治疗方法显示出强大的抗肿瘤效果。CPMV是一种肿瘤内病变 注射原位疫苗(ISV)技术，具有诱导持久免疫介导性抗肿瘤的能力 对已治疗和未治疗的肿瘤的疗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，以改善肿瘤相关和 新抗原，从而产生更有功能的适应性抗肿瘤免疫，对抗由 肿瘤。这导致已治疗和未治疗的恶性肿瘤得到缓解，并防止复发 通过适应性手臂和免疫记忆。建立在数据和知识的坚实基础上 性质，这个项目将专注于优化GMP的制造和在小鼠肿瘤模型中的药理 为今后的人类肿瘤学试验提供基础。多元PI领导 斯坦梅茨团队(加州大学圣地亚哥分校)、菲林(达特茅斯分校)和兰詹(俄亥俄州立大学)带来了免费的专业知识 以植物病毒为基础的CPMV纳米技术、癌症免疫学和兽医肿瘤学/免疫疗法试验。 我们的团队还得到了一组在监管方面具有专业知识的顾问的支持(Garnick，Mosaic IE Inc.)， 药理学/毒理学(Luksic，Intrinsik)，临床(Garoway，Mosaic IE Inc.)事务，以及经验 在垂直农业(Eisenberg，OPO)。我们相信，我们强大的合作者团队可以帮助建立 未来植物衍生生物的制造。我们将实现以下具体目标：(1)化学， 用于CPMV可扩展制造和QAC的制造和控制(CMC)方法开发- ISV，(2)药理学：确定CPMV-ISV候选药物的生物分布、剂量和毒理学 黑色素瘤的原位和转移性小鼠模型；以及(3)犬试验：建立剂量和安全性 适用于原位接种的不同肿瘤类型(黑色素瘤、肉瘤和肥大)的犬肿瘤学试验 细胞瘤)。通过对小鼠肿瘤模型和狗患者的平行试验，我们将识别活组织检查或血液 用于纵向监测治疗反应的生物标志物。如果成功完成，我们将永远处于有利地位 生产实践(GMP)这种以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 以CPMV为基础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.