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 该提案旨在建立在我们在犬免疫治疗试验中的广泛初步数据的基础上，以证明 外源性细胞因子递送与自体自然杀伤（NK）细胞组合的新方法， 首次在犬中进行的临床试验代表了一种潜在的高影响力方法，可优化基于非T细胞的 免疫疗法这项工作的目标是狗与典型的致命性，自然发生的骨肉瘤（OSA） 和黑色素瘤，这些发现预计将对设计和翻译具有重要意义。 创新的人类免疫治疗方法。吸入（IH）IL-15提供局部递送IL-15的优点。 这种免疫刺激细胞因子，同时限制全身暴露，从而限制潜在的毒性。超激动剂IL- 15提供增加的半衰期和更大的抗肿瘤作用。因此，预期这些细胞因子中的每一种都 改变风险/收益比以利于其使用。为实现这些目标，我们提出以下三点建议： 具体目的：1）靶向大体肺转移（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IL-15的II期试验 总OSA和黑色素瘤肺转移，和; 3）使用超- IL-15激动剂在原发性OSA中的作用。犬模型是癌症免疫治疗研究的有力工具 作为小鼠模型和人类临床研究之间的重要联系。狗代表了一种有吸引力的远系繁殖动物 在完整的环境中经历自发性癌症发展的伴侣动物的组合 免疫系统重要的是，这些研究旨在评估新的治疗组合， 晚期肺转移瘤在肉眼病变的情况下，也将决定这一影响 当与标准治疗相结合时，将其作为一线治疗。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， 患有自然发生的癌症的狗。我们将有独特的机会来展示整个外显子组 测序和RNASeq对免疫治疗前后肿瘤组织的影响，我们将研究是否存在免疫治疗。 可以开发突变景观和基因表达谱特征以前瞻性地预测 结果和对免疫疗法的反应。我们将这些数据与免疫组化、流式细胞术、 从“冷”到“热”肿瘤转变的证据的血清细胞因子读数， 有利的长期结果。因此，这些犬试验和生物标记物研究代表了一种理想的策略， 为人类患者提供信息并促进新型、潜在高影响免疫疗法的快速转化 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.