Gene Expression Analysis of Dog Natural Killer Cells as Immunotherapy Target

作为免疫治疗靶标的狗自然杀伤细胞的基因表达分析

• 批准号: 10201901
• 负责人: Robert J. Canter
• 金额: $ 7.85万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10201901
• 关键词: Address; Adoptive Transfer; Animals; Antigen-Antibody Complex; Bioinformatics; Biological; Biological Assay; Biology; Blood; Cancer Patient; Candidate Disease Gene; Canis familiaris; Cells; Cellular biology; Clinical; Clinical Trials; Coculture Techniques; Collaborations; Data; Data Analyses; Development; Dissection; Environmental Exposure; Evaluation; Flow Cytometry; Future; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Genes; Genetic; Genomics; Goals; Growth; High-Throughput Nucleotide Sequencing; Homing; Human; Human Genome; Human Microbiome; Immune; Immune system; Immunologic Markers; Immunology; Immunooncology; Immunotherapy; Inhalation; Interleukin-15; Investigation; K562 Cells; Knowledge; Light; Link; Longevity; Malignant Neoplasms; Membrane; Metagenomics; Metastatic Osteosarcoma; Modeling; Mus; NK cell therapy; Natural Killer Cells; Ontology; Paper; Pathway interactions; Phenotype; Principal Component Analysis; Production; Progressive Disease; Reagent; Recording of previous events; Research; Rest; Role; Sampling; Sequence Analysis; Specimen; Speed; Techniques; Technology; Tissue-Specific Gene Expression; Tissues; Toxic effect; Translations; Veterinary Medicine; Veterinary Schools; Visual; base; bench to bedside; cancer cell; cancer immunotherapy; cancer therapy; clinical predictors; clinical translation; companion animal; comparative; cytokine; cytotoxicity; differential expression; dog genome; exhaustion; genetic signature; improved; in vivo; medical schools; mouse model; neoplastic cell; novel; osteosarcoma; overexpression; pre-clinical; preclinical study; predictive signature; premature; receptor; responders and non-responders; response; sarcoma; single-cell RNA sequencing; species difference; targeted treatment; trait; transcriptome; transcriptome sequencing; transcriptomics; translational model; tumor; tumor immunology

Project Summary Dogs are large, outbred animals that develop cancer spontaneously in the presence of an intact immune system and in the setting of shared environmental exposures with humans. Humans and dogs also 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. 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 high impact studies using dog immunotherapy are premature until a deeper understanding of dog immunology and immune biomarkers is realized, especially for natural killer (NK) cells. This proposal will build on our exciting preliminary data characterizing dog NK cells, including first-in-dog clinical trials of adoptive transfer of dog NK cells and novel cytokine delivery with inhaled IL-15. In collaboration with the School of Medicine, the School of Veterinary Medicine, and the Data Intensive Biology Lab which performs cross-species genomic, transcriptomic, and metagenomic sequence analyses, we will examine dog NK differential gene expression across 1) multiple stimulatory and steady-state conditions; 2) from dog blood and tumor bio-specimens; and 3) from responding and non-responding dogs treated with inhaled IL-15 on an investigational dog clinical trial. We will utilize multiple applications of high-throughput sequencing technologies (RNASeq and single cell RNA sequencing) to establish more robust and validated gene signatures of resting and activated dog NK cells from both ex vivo and in vivo conditions with further analysis of dog sarcoma-infiltrating NK cells. We will then compare these gene signatures to data from high throughput sequencing of murine and human NK cells to bridge species differences and facilitate clinical translation. Ultimately, knowledge of the role of canine NK cells in immune defense and cancer therapy is limited, and it remains unclear whether this is due to biological differences across species or limited reagents for investigation. Results from this proposal will extend the important link that canine studies provide between murine pre-clinical studies and human clinical trials, for cancer immunotherapy research in general but especially for NK studies where high impact clinical translation has remained elusive.

项目摘要 狗是大型的大脑动物，在完整的免疫系统存在下会自发发展癌症 并在与人类的共同环境暴露环境中。人类和狗也有一对 进化史导致犬与人类基因组和微生物组之间的相似之处更大 比老鼠和人之间。这些特征共同使狗成为有利的翻译模型 研究癌症免疫学和癌症免疫疗法。狗临床试验允许研究复杂的免疫 治疗过程中的相互作用，同时还解决了癌症免疫疗法的长期疗效和毒性。 但是，免疫解剖需要开发可靠和经过验证的测定法和试剂，并且很高 使用狗免疫疗法的撞击研究为时过早，直到对狗免疫学和 实现了免疫生物标志物，特别是对于天然杀手（NK）细胞。该提议将基于我们令人兴奋的 表征狗NK细胞的初步数据，包括狗NK的收养转移的第一试验试验 细胞和新型的细胞因子递送使用吸入的IL-15。与医学院合作 兽医医学以及执行跨物种基因组，转录组的数据密集型生物学实验室 和宏基因组序列分析，我们将检查1）多个的狗NK NK差异基因表达 刺激和稳态条件； 2）来自狗血和肿瘤生物特异性； 3）回应 在研究犬临床试验中，用吸入IL-15治疗的无反应犬。我们将使用多个 高通量测序技术（RNASEQ和单细胞RNA测序）的应用以建立 来自体内和体内的静止和激活的狗NK细胞的更稳定和验证的基因特征 通过进一步分析狗肉瘤浸润的NK细胞的条件。然后，我们将比较这些基因特征 到来自鼠和人类NK细胞高吞吐量测序的数据，以弥合物种差异和 促进临床翻译。最终，了解犬NK细胞在免疫防御和癌症中的作用 治疗是有限的，尚不清楚这是由于物种之间的生物学差异或有限 调查试剂。该提案的结果将扩展犬类研究提供的重要链接 在鼠前研究和人类临床试验之间，一般进行癌症免疫疗法研究，但 特别是对于高影响临床翻译仍然难以捉摸的NK研究。