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

摘要 这份R01续签申请专注于三重阴性乳腺癌(TNBC)，这是一种侵略性的生活- 预后差，复发和远处转移的可能性增加的威胁疾病。 癌症免疫治疗的进展表明，对患者免疫系统的调节可以 具有显著的抗肿瘤活性。最有希望的免疫治疗方法是那些 个性化并利用每个患者肿瘤中独特的新抗原。为了实现这个目标，我们 开发了一种植物病毒纳米颗粒免疫疗法，可以激活体内的天然免疫细胞 肿瘤微环境(TME)以启动适应性、系统性和持久的抗肿瘤免疫。 具体地说，瘤内注射豌豆花叶病毒(CPMV)在多只小鼠身上显示出强大的疗效 型号，包括TNBC。在患有乳腺癌的伴侣犬身上的试验也表明了强大的抗肿瘤效果。 在前一个供资周期中，我们对行动机制有了深入的了解，并证明 CPMV由病原体相关分子模式(PAMP)受体识别，PAMP受体检测危险信号 并激活先天免疫系统；具体地说，CPMV由Toll样受体(TLR2、4和7)识别。 此外，我们开发并测试了组合和双管齐下的治疗方法：我们演示了 CPMV单独治疗及与放化疗、免疫调节联合治疗的疗效观察 药物和检查站抑制剂等。这项建议建立在这一强大的数据组合之上。我们的第一次 目标是专注于结合了免疫调节和抗肿瘤免疫特性的双管齐下的CPMV 使用检查点疗法(目标1)。检查点阻断抗体在消除抑制信号方面是有效的，但 AS单一疗法疗效多变且有限。CPMV原位免疫可增加肿瘤抗原 特异性效应T细胞和我们的初步数据表明，CPMV治疗与免疫协同作用 检查点疗法。接下来，我们寻求开发有针对性的方法，有效地系统地集中精力 在肿瘤中实施CPMV，并为治疗转移性疾病提供进一步的治疗选择(目标2)。S100A9- 将研究靶向CPMV：S100A9(也称为髓系相关蛋白14[MRP-14])的表达，是 与炎症和癌症有关。S100A9在乳腺癌中的高表达与病情恶化相关 预后。S100A9的表达是肿瘤发生、增强肿瘤侵袭性和 转移。在TME中，S100A9被分泌到细胞外基质，使其成为非常适合的靶点 纳米医学。认识到S100A9作为药理靶点的潜力，我们开发了靶向S100A9 有效地集中在转移部位的CPMV，能够有效地防止肿瘤的生长 转移瘤。在这里，我们将详细介绍S100A9的作用机制，并了解其药理作用。 靶向CPMV。最后，随着兽医临床试验的进行，临床前开发将大幅延长 患有乳腺肿瘤的伴侣犬(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).