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: 10372182
• 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 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10372182
• 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; 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; canine model; 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.

项目概要 狗是大型远交动物，在免疫系统完整的情况下会自发患上癌症 以及与人类共同暴露于环境的环境中。人类和狗也有配对 进化史导致犬类和人类基因组和微生物组之间存在更大的相似性 比老鼠和人类之间。总之，这些特征使狗成为一种有利的转化模型 研究癌症免疫学和癌症免疫治疗。狗临床试验可以研究复杂的免疫 治疗期间的相互作用，同时还解决癌症免疫疗法的长期疗效和毒性。 然而，免疫解剖需要开发强大且经过验证的检测方法和试剂，以及高 在对狗的免疫学有更深入的了解之前，使用狗免疫疗法进行影响研究还为时过早 免疫生物标志物被实现，特别是对于自然杀伤（NK）细胞。该提案将建立在我们令人兴奋的基础上 狗 NK 细胞特征的初步数据，包括狗 NK 过继转移的首次狗临床试验 细胞和吸入 IL-15 的新型细胞因子递送。与医学院合作， 兽医学和数据密集型生物学实验室，该实验室进行跨物种基因组、转录组、 和宏基因组序列分析，我们将检查狗 NK 差异基因表达：1) 多个 刺激和稳态条件； 2）来自狗血和肿瘤生物样本； 3）来自回应 以及在一项狗临床试验中接受吸入 IL-15 治疗但无反应的狗。我们将利用多个 应用高通量测序技术（RNASeq和单细胞RNA测序）建立 来自离体和体内的静息和激活的狗 NK 细胞的更强大和经过验证的基因特征 进一步分析狗肉瘤浸润的 NK 细胞。然后我们将比较这些基因特征 来自小鼠和人类 NK 细胞高通量测序的数据，以弥合物种差异 促进临床转化。最终，了解犬类 NK 细胞在免疫防御和癌症中的作用 治疗是有限的，目前尚不清楚这是由于物种间的生物学差异还是有限的 用于研究的试剂。该提案的结果将扩展犬类研究提供的重要联系 小鼠临床前研究和人体临床试验之间，一般用于癌症免疫治疗研究，但 尤其是对于 NK 研究，高影响力的临床转化仍然难以实现。

项目成果

Using the canine microbiome to bridge translation of cancer immunotherapy from pre-clinical murine models to human clinical trials.

利用犬微生物组将癌症免疫疗法从临床前小鼠模型转化为人体临床试验。

• 发表时间: 2022
• 作者: Kleber, Kara T;Iranpur, Khurshid R;Perry, Lauren M;Cruz, Sylvia M;Razmara, Aryana M;Culp, William T N;Kent, Michael S;Eisen, Jonathan A;Rebhun, Robert B;Canter, Robert J
• 通讯作者: Canter, Robert J