Overcoming the immune-suppressive tumor microenvironment through in situ vaccination nanotechnology.

总结 肿瘤产生的免疫抑制微环境是免疫清除的关键屏障 从而使癌症得以显现。我们建议使用植物病毒样纳米棒来克服这一障碍， 原位疫苗接种的颗粒（VLP）平台技术，以消除局部免疫抑制并产生有效的免疫抑制。 有效的局部和全身抗肿瘤免疫。我们最近证明了植物生产的，工程化的 基于VLP的纳米技术在转移性肿瘤小鼠模型中刺激有效的抗肿瘤免疫应答 黑色素瘤卵巢癌和乳腺癌数据表明，这种影响是系统性和持久的，导致 免疫记忆和防止复发。转移性癌伴犬的初步研究 黑色素瘤的研究表明，有效的抗肿瘤功效可以在犬模型中复制，该模型具有高 与人类黑色素瘤有关原位疫苗接种提供了一种个性化的治疗方法， 患者的肿瘤介导的免疫抑制和增强针对抗原的抗肿瘤免疫， 被自己的肿瘤压迫使用植物VLP的拟议纳米工程方法将改善 护理标准以多种方式。首先，原位接种会增加抗原特异性T细胞的频率， 细胞，导致持久的免疫记忆;这与检查点抑制剂相反，检查点抑制剂不是 抗原特异性，激活所有抗原经历的T细胞并导致脱靶免疫毒性。其次，FDA- 批准的使用T-VEC的原位疫苗接种证明了该方法，但由于使用at- 具有传染性的减弱疱疹病毒稳定性差，每剂价格昂贵。而 我们的支持数据表明，在各种肿瘤模型中， 非常独特，尚未完全理解。因此，这一提议的实质是要破译 强大的纳米技术的工程设计空间，并描绘了潜在的机制， 行动主义。因此，本研究的目的是：1）破译VLP的分子结构; 触发强大功效的更大特征。2)阐明免疫激活的潜在机制 由贵宾准备的3)通过研究，进一步了解其作用机制和疗效， 患有口腔黑素瘤的伴侣犬。多PI伙伴关系和合作研究者将有助于 该计划的成功。总之，数据将提供抗肿瘤免疫激活的详细机制 通过工程VLP纳米技术，并将通知纳米工程，以进一步改进的方法， 开始在人类患者身上进行测试。

Summary The immune-suppressive microenvironment generated by tumors is a key barrier to immune clearance and thereby, enables cancer to manifest. We propose to overcome this barrier using a plant virus-like nanopar- ticle (VLP) platform technology for in situ vaccination, to eliminate local immunosuppression and generate ef- fective local and systemic anti-tumor immunity. We recently demonstrated that plant-produced, engineered VLP-based nanotechnologies stimulate a potent anti-tumor immune response in mouse models of metastatic melanoma, ovarian cancer, and breast cancer. Data indicate that the effect is systemic and durable, resulting in immune-memory and protection from recurrence. Preliminary studies in companion dogs with metastatic melanoma indicate that the potent anti-tumor efficacy can be replicated in the canine model, which has high relevance to human melanoma. In situ vaccination provides a personalized treatment approach by relieving the patient's tumor-mediated immunosuppression and potentiating anti-tumor immunity against antigens ex- pressed by their own tumor. The proposed nanoengineering approach using plant VLPs would improve the standard of care in several ways. First, in situ vaccination will increase the frequency of antigen-specific T cells, leading to long-lasting immunologic memory; this is in contrast to checkpoint inhibitors, which are not an- tigen specific, activate all antigen-experienced T cells and result in off-target immune toxicities. Second, FDA- approved in situ vaccination using T-VEC demonstrates the approach but is limited because of the use of at- tenuated herpes virus, which can be infectious, has poor stability and an extraordinary price per dose. While our supporting data indicate potent efficacy in various tumor models, the underlying immunology is quite unique and not yet completely understood. Therefore, the essence of this proposal is to decipher the engineering design space of the potent nanotechnology and to delineated the underlying mecha- nism of action. Therefore, this proposal sets out to fulfill the following aims: 1) To decipher the VLP's molecu- lar features triggering the potent efficacy. 2) To delineate the underlying mechanisms of immune activation primed by the VLPs. 3) To gain further insights into the mechanism of action and efficacy through study of companion dogs with oral melanoma. A multi-PI partnership and collaborating investigators will contribute to the success of the program. Together, data will provide detailed mechanisms of anti-tumor immune activation by engineered VLP nanotechnologies and will inform nanoengineering to further improve the approach and begin the effort to test it in human patients.