Enhancing natural killer immunotherapy with first-in-dog trials of inhaled recombinant IL-15 and super-agonist IL-15 in naturally occurring canine cancers

项目摘要 这项提案寻求建立在我们在犬类免疫治疗试验中的广泛初步数据的基础上，以证明 外源性细胞因子与自体自然杀伤(NK)细胞相结合的新方法 首个狗内临床试验代表了一种潜在的高影响力的方法，以优化基于非T细胞的 免疫疗法。这项研究的目标是患有典型的致命的自然发生的骨肉瘤(OSA)的狗。 和黑色素瘤，预计这些发现将对设计和翻译 创新的人类免疫治疗方法。吸入型(IH)IL-15提供局部给药的优势 这种免疫刺激细胞因子，同时限制全身暴露，从而潜在的毒性。超级激动剂IL- 15具有更长的半衰期和更强的抗肿瘤作用。因此，预计这些细胞因子中的每一种都可以 改变风险/收益比率，以利于它们的使用。为了实现这些目标，我们提出了以下三项建议 具体目标：1)通过第一次犬内交付靶向肉眼肺转移(OSA和黑色素瘤) 吸入(IH)人IL-15和超激动剂IL-15；2)IH IL-15和自体NK细胞治疗的II期试验 大体OSA和黑色素瘤肺转移，以及；3)针对微转移疾病使用超 原发阻塞性睡眠呼吸暂停综合征患者使用激动剂IL-15。犬类模型是癌症免疫治疗研究的有力工具。 作为连接小鼠模型和人类临床研究的重要纽带。狗代表着有吸引力的近亲繁殖 在完整的环境中经历自发癌症发展的同伴动物的组合 免疫系统。重要的是，这些研究旨在评估新的治疗组合 在晚期肺转移的背景下，大体疾病，也将决定这一影响 当与标准护理治疗相结合时，将该方法作为一线治疗。此外，这项工作旨在 阐明介导抗肿瘤反应(或无反应)的细胞和分子机制 患有自然癌症的狗。我们将有一个独特的机会来表演整个exome 对免疫治疗前后的肿瘤组织进行测序和RNAseq，我们将调查 突变景观和基因表达谱的特征可以被开发来前瞻性地预测 免疫治疗的结果和反应。我们会将这些数据与免疫组织化学、Flow 细胞学和血清细胞因子读数作为肿瘤从冷态向热态转变的证据 有利的长期结果。因此，这些犬类试验和生物标记研究代表了一种理想的策略 告知并促进将新型的、潜在的高影响免疫疗法快速翻译给人类患者 患有侵袭性癌症。

Project Summary This proposal seeks to build on our extensive preliminary data in canine immunotherapy trials to demonstrate that novel methods of exogenous cytokine delivery in combination with autologous natural killer (NK) cells in first-in-dog clinical trials represents a potentially high impact approach to optimize non-T cell based immunotherapies. This work is targeted to dogs with typically lethal, naturally occurring osteosarcoma (OSA) and melanoma, and the findings are expected to have important relevance for the design and translation of innovative immunotherapy approaches in humans. Inhaled (IH) IL-15 offers the advantages of local delivery of this immune-stimulatory cytokine, while limiting systemic exposure and thus potential toxicity. Super-agonist IL- 15 offers increased half-life and greater anti-tumor effects. Therefore, each of these cytokines is anticipated to alter the risk/benefit ratio in favor of their use. To accomplish these objectives, we propose the following three specific aims: 1) Targeting gross pulmonary metastases (OSA and melanoma) with first-in-dog delivery of inhaled (IH) human IL-15 and super-agonist IL-15; 2) Phase II trial of IH IL-15 and autologous NK cells to treat gross OSA and melanoma pulmonary metastases, and; 3) Targeting micro-metastatic disease using super- agonist IL-15 in primary OSA. The canine model represents a powerful tool in cancer immunotherapy research as an important link between murine models and human clinical studies. Dogs represent an attractive outbred combination of companion animals that experience spontaneous cancer development in the setting of an intact immune system. Importantly, these studies are designed to evaluate novel treatment combinations for advanced pulmonary metastases in the setting of gross disease, but also will determine the impact of this approach as first-line therapy when combined with standard-of-care treatments. Moreover, this work seeks to elucidate the cellular and molecular mechanisms that mediate an anti-tumor response (or non-response) in dogs with naturally occurring cancers. We will have the unique opportunity to perform whole exome sequencing and RNASeq on tumor tissue pre- and post-immunotherapy, and we will investigate whether a signature of mutational landscape and gene expression profiles can be developed to prospectively predict outcome and response to immune therapies. We will correlate these data to immunohistochemical, flow cytometry, and serum cytokine readouts for evidence of transition from “cold” to “hot” tumors predictive of favorable long term outcome. Hence, these canine trials and bio-marker studies represent an ideal strategy to inform and facilitate the rapid translation of novel, potentially high impact immune therapies to human patients with aggressive cancers.