Dual-pronged nano-drug delivery using plant virus-like particles

总结 本次R 01续期申请重点关注三阴性乳腺癌（TNBC），这是一种侵略性的生活- 预后差、复发和远处转移可能性增加的威胁性疾病。 癌症免疫治疗的进展已经证明，调节患者的免疫系统可以 导致显著的抗肿瘤活性。最有前途的免疫治疗方法是那些 个性化并利用每个患者肿瘤内的独特新抗原。为了实现这一目标，我们 开发了一种植物病毒纳米颗粒免疫治疗方法， 肿瘤微环境（TME）启动适应性，系统性和持久的抗肿瘤免疫。 具体地，肿瘤内注射豇豆花叶病毒（CPMV）在多个小鼠中显示出有效的功效。 模型，包括TNBC。在患有乳腺癌的伴侣犬中进行的试验也证明了有效的抗肿瘤疗效。 在上一个供资周期，我们深入了解了作用机制，并证明， CPMV被病原体相关分子模式（PAMP）受体识别，该受体检测危险信号 并激活先天免疫系统;具体而言，CPMV被Toll样受体（TLR 2、4和7）识别。 此外，我们开发并测试了联合和双管齐下的治疗方法：我们证明了 CPMV单独治疗以及与放疗、化疗、免疫调节剂联合治疗疗效 药物和检查点抑制剂等。这一建议建立在这一强大的数据组合基础上。我们的第一 我们的目标是集中在双管齐下的CPMV，结合其免疫调节和抗肿瘤免疫特性 检查点疗法（目标1）。检查点阻断抗体可有效去除抑制信号， 因为单一疗法的疗效可变且有限。原位接种CPMV增加肿瘤抗原 特异性效应T细胞和我们的初步数据表明，CPMV治疗与免疫应答协同作用， 检查点疗法接下来，我们寻求制定有针对性的方法，有效地集中系统 在肿瘤中施用CPMV并提供进一步的治疗选择以治疗转移性疾病（目的2）。S100A9- 将研究靶向CPMV：S100 A9（也称为骨髓相关蛋白14 [MRP-14]）的表达， 与炎症和致癌有关。乳腺癌中S100 A9的高表达与乳腺癌的预后相关。 预后S100 A9表达是肿瘤发生的早期事件，增强了肿瘤的侵袭性， 转移在TME中，S100 A9分泌到细胞外基质，使其成为高度适合的靶点， 纳米医学认识到S100 A9作为药理学靶点的潜力，我们开发了S100 A9靶向 CPMV有效地集中在转移部位，从而能够有效地防止肿瘤的生长。 转移在这里，我们开始详细介绍S100 A9的作用机制和药理学。 针对CPMV。最后，临床前开发将通过兽医临床试验得到实质性扩展 在患有乳腺肿瘤的伴侣犬中（目标3）。

Summary This R01 renewal application is focused on triple negative breast cancer (TNBC), which is an aggressive life- threatening disease with poor prognosis and increased likelihood of recurrence and distant metastasis. Advances in cancer immunotherapy have demonstrated that modulation of the patient’s immune system can result in dramatic antitumor activity. The most promising immunotherapy approaches are those that are personalized and take advantage of the unique neoantigens within each patient’s tumor. Toward this goal, we developed a plant virus nanoparticle immunotherapy approach that activates innate immune cells within the tumor microenvironment (TME) to launch adaptive, systemic, and durable antitumor immunity. Specifically, intratumorally injected cowpea mosaic virus (CPMV) demonstrates potent efficacy in multiple mouse models, incl. TNBC. Trials in companion dogs with breast cancer also demonstrate potent antitumor efficacy. During the previous funding cycle, we gained insights into the mechanism of action and demonstrated that CPMV is recognized by pathogen-associated molecular pattern (PAMP) receptors that detect danger signals and activate the innate immune system; specifically, CPMV is recognized by Toll-like receptors (TLR2, 4 and 7). Further, we developed and tested combination and dual-pronged treatment approaches: we demonstrated efficacy of CPMV as solo-treatment as well as in combination with radiation, chemotherapy, immunomodulatory drugs, and checkpoint inhibitors, amongst others. This proposal builds on this strong portfolio of data. Our first goal is to focus on dual-pronged CPMV that combines its immunomodulatory and antitumor immunity properties with checkpoint therapy (Aim 1). Checkpoint blocking antibodies are effective at removing inhibitory signals but as monotherapy have variable and limited efficacy. In situ vaccination with CPMV increases tumor antigen specific effector T cells and our preliminary data indicate that CPMV treatment synergizes with immune checkpoint therapy. Next, we seek to develop targeted approaches that effectively concentrate systemically administered CPMV in tumors and provide further therapy options to treat metastatic disease (Aim 2). S100A9- targeted CPMV will be studied: expression of S100A9 (also known as myeloid-related protein 14 [MRP-14]), is linked to inflammation and carcinogenesis. Higher S100A9 expression in breast cancer correlates with a worse prognosis. S100A9 expression is an early event in tumorigenesis, enhancing tumor aggressiveness and metastasis. In the TME, S100A9 is secreted to the extracellular matrix, making it a highly suitable target for nanomedicine. Recognizing the potential of S100A9 as a pharmacologic target, we developed S100A9-targeted CPMV that efficiently concentrates at sites of metastasis enabling potent efficacy preventing outgrowth of metastases. Here we set out to detail the mechanisms of action and understand the pharmacology of S100A9- targeted CPMV. Finally, the preclinical development will be substantially extended with veterinary clinical trials in companion dogs with mammary tumors (Aim 3).