Gene Expression Analysis of Dog Natural Killer Cells Following Immunotherapy with Molecularly Targeted Radionuclide therapy or Inhaled IL-15

项目摘要 伴侣犬是大型的远系繁殖动物，在存在癌症的情况下自发地发展癌症。 一个完整的免疫系统，一个原生的肿瘤微环境， 与人类共享的环境暴露。事实上，人类和狗共享一对 进化史导致了犬类和人类基因组之间更大的相似性 和微生物组之间的差异。总之，这些特征使狗成为一个 研究癌症免疫学和癌症免疫治疗的有利的转化模型，以及 狗的临床试验允许在治疗期间研究复杂的免疫相互作用， 解决癌症免疫疗法的长期功效和毒性。然而，免疫 解剖需要开发稳健和有效的测定和试剂，以及更深入的 了解狗的免疫学和免疫生物标志物是必要的，以实现高的影响 翻译研究，特别是在自然杀伤（NK）细胞的背景下， 已知存在物种差异。这项提案将建立在令人兴奋的数据， 合作机构在比较肿瘤学和新的 免疫治疗试验正在进行中。使用来自试验犬的治疗前和治疗中PBMC样本 接受新的和“一流的”免疫疗法，刺激NK细胞，我们将使用RNA 测序以检查NK差异基因表达，以1）建立稳健且经验证的 来自两个临床试验群组的静息和活化的狗NK细胞的基因特征; 2）分析 来自应答和非应答患者的NK基因标签;和3）评估NK基因标签的特异性。 不同NK刺激因子之间NK基因特征的相似性和差异 免疫疗法预期具有重叠但不同的作用机制。 这一提议的结果将扩展犬研究提供的重要联系， 小鼠临床前研究和人类临床试验，并促进新的， 对患有侵袭性癌症的狗和人类患者的潜在高影响NK疗法。

Project Summary Companion dogs are large, outbred animals that develop cancer spontaneously in the presence of an intact immune system, a native autochthonous tumor microenvironment, and in the setting of shared environmental exposures with humans. In fact, humans and dogs share a paired evolutionary history which has led to greater similarities between canine and human genomes and microbiomes than between mouse and human. Together, these traits make dogs an advantageous translational model to study cancer immunology and cancer immunotherapy, and dog clinical trials allow for the study of complex immune interactions during therapy while also addressing long-term efficacy and toxicity of cancer immunotherapies. However, immune dissection requires the development of robust and validated assays and reagents, and a deeper understanding of dog immunology and immune biomarkers is necessary to achieve high impact translational studies, especially in the context of natural killer (NK) cells where significant species differences are known to exist. This proposal will build on exciting data from the collaborating institutions with significant expertise in comparative oncology and novel immunotherapy trials underway. Using pre- and on-treatment PBMC samples from dogs on trial receiving novel and “first-in-class” immunotherapies which stimulate NK cells, we will use RNA sequencing to examine NK differential gene expression to 1) establish robust and validated gene signatures of resting and activated dog NK cells from both clinical trial cohorts; 2) analyze NK gene signatures from responding and non-responding patients; and 3) evaluate for similarities and differences in NK gene signatures between respective NK stimulatory immunotherapies which are anticipated to have overlapping, but distinct, mechanisms of action. Results from this proposal will extend the important link that canine studies provide between murine pre-clinical studies and human clinical trials and facilitate the rapid translation of novel, potentially high impact NK therapies to both dog and human patients with aggressive cancers.