Examining the impact of laboratory housing temperature on murine CD28 and the response to anti-PD-1

检查实验室外壳温度对小鼠 CD28 的影响以及抗 PD-1 的反应

基本信息

批准号：
10751424
负责人：
Caitlin M James
金额：
$ 3.25万
依托单位：
ROSWELL PARK CANCER INSTITUTE CORP
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10751424
关键词：
Address Affect Animal Housing Animal Model Animals Basal metabolic rate Biomedical Research CD28 gene CD8-Positive T-Lymphocytes CD8B1 gene Cancer Model Caring Chronic Clinic Clinical Data Development Down-Regulation Flow Cytometry Genetically Engineered Mouse Goals Guidelines Harvest House mice Housing Human Immune Immune checkpoint inhibitor Immunologics Immunotherapy Impairment In Vitro Institution Knockout Mice Knowledge Laboratories Laboratory Animal Science Laboratory Animals Malignant Neoplasms Measures Mediating Medical Modeling Molecular Molecular Target Mus Nerve Norepinephrine Outcome Pathway interactions Patients Phenotype Physiological Play Pre-Clinical Model Process Publishing Receptor Signaling Reproducibility Research Research Proposals Rodent Role Signal Induction Signal Transduction Standardization Surface Sympathetic Nervous System T-Cell Activation T-Lymphocyte Temperature Testing Thermogenesis Training Tumor Immunity Tumor-Infiltrating Lymphocytes Work adrenergic stress anti-PD-1 anti-tumor immune response anticancer research beta-adrenergic receptor cancer therapy career cold stress design effector T cell environmental stressor experience falls functional disability guide for the care and use of laboratory animals human disease human model improved in vivo insight mRNA Expression melanoma mouse model novel therapeutics pre-clinical preclinical study protein expression receptor response success therapy development translational potential treatment response tumor tumor growth tumor immunology

项目摘要

Project Abstract/Summary For decades, animals in biomedical research have yielded significant scientific and medical breakthroughs by generating the essential preclinical data that ultimately support the discovery and development of treatments for human diseases, including cancer. However, while we to rely on animal models to investigate the complexity of cancer and cancer therapies, these preclinical studies have alarmingly low success in reproducibility, and even lower preclinical-to-clinical success rates. As per the Guide for the Care and Use of Laboratory Animals 8th Edition, research institutions have standardized, minimum guidelines for the housing, husbandry, and overall care for laboratory animals that they must adhere to. A mildly cool ambient temperature is a critical aspect of animal housing that has been shown to elicit significant physiological changes to research rodents, driven by the activation of the sympathetic nervous system and increased β- adrenergic receptor (β-AR) signaling as a result of the systemic release of norepinephrine. This is due to the compensatory response, known as non-shivering thermogenesis, employed by rodents housed at temperatures that fall below their thermoneutral zone (which is the range of ambient temperatures at which heat generated by basal metabolism is sufficient for maintaining homeostatic core temperature. ) Our lab has previously established that standard (ST), subthermoneutral laboratory housing temperatures result in significant impairment to the murine CD8+ T cell-dependent anti-tumor immune response compared to mice house at thermoneutral temperatures (TT). Additionally, we have shown that the immune checkpoint inhibitor αPD-1, an immunotherapy that has recently seen success as a front-line approach to treating cancers like melanoma, has improved efficacy in treating tumor-bearing mice housed at TT in a β-AR-dependent manor. Although published and preliminary data indicate a role for the co-receptor, CD28, in the diminished anti-tumor function of CD8+ T cells as a result of increased β-AR signaling, a gap exists in our understanding of the mechanisms underlying the reduced CD8+ T cell activation and effector function in mice housed at ST. Therefore, we propose using genetically engineered mouse models to precisely interrogate CD28 signaling and test hypothesis that standard housing temperatures impairs CD8+ T cell anti-tumor immunity and the in vivo efficacy of αPD-1 via impaired CD28 co-stimulation. We will use in vitro and in vivo approaches to examine the effects of housing temperature on CD8+ T cell CD28 expression and signaling, as well as tumor- infiltrating lymphocytes in mice treated with αPD-1 therapy. The studies outline in this proposal have the potential to identify a previously undefined mechanism by which subthermoneutral laboratory animal housing temperatures influence experimental outcomes of cancer and immunotherapy models, while also characterizing a widely underappreciated variable that exists in our animal models.

项目摘要/总结几十年来，生物医学研究中的动物取得了重大的科学和医学突破：生成最终支持治疗方法的发现和开发的重要临床前数据然而，虽然我们要依靠动物模型来研究人类疾病，包括癌症。由于癌症和癌症治疗的复杂性，这些临床前研究的成功率低得惊人重现性，甚至更低的临床前到临床成功率。实验动物第 8 版，研究机构有标准化的、最低限度的住房指南，饲养和对实验动物的整体护理，它们必须遵守温和凉爽的环境。温度是动物圈舍的一个重要方面，已被证明会引起显着的生理影响研究啮齿类动物的变化是由交感神经系统的激活和β-增加所驱动的肾上腺素受体（β-AR）信号传导是去甲肾上腺素全身释放的结果。补偿性反应，称为非颤抖产热，由饲养在温度低于其热中性区（即环境温度范围）基础代谢产生的热量足以维持体内平衡的核心温度）我们的实验室有。先前确定标准 (ST)、亚热中性实验室外壳温度会导致与小鼠相比，小鼠 CD8+ T 细胞依赖性抗肿瘤免疫反应显着受损此外，我们还证明了免疫检查点抑制剂。 αPD-1，一种免疫疗法，最近作为治疗癌症的一线方法取得了成功黑色素瘤，对饲养在 TT β-AR 依赖性庄园中的荷瘤小鼠的治疗效果有所提高。尽管已发表的初步数据表明辅助受体 CD28 在抗肿瘤作用减弱中发挥作用由于 β-AR 信号传导增强，CD8+ T 细胞的功能受到影响，但我们对 CD8+ T 细胞功能的理解存在差距。 ST 小鼠 CD8+ T 细胞活化和效应器功能减少的机制。因此，我们建议使用基因工程小鼠模型来精确询问 CD28 信号传导并测试标准外壳温度会损害 CD8+ T 细胞抗肿瘤免疫力的假设以及通过受损的 CD28 共刺激来验证 αPD-1 的体内功效我们将使用体外和体内方法来研究。检查外壳温度对 CD8+ T 细胞 CD28 表达和信号传导以及肿瘤的影响接受 αPD-1 治疗的小鼠体内的浸润淋巴细胞本提案中的研究概述如下。有可能确定以前未定义的机制，通过该机制，亚温中性的实验动物住房温度影响癌症和免疫治疗模型的实验结果，同时也会影响描述我们动物模型中存在的一个被广泛低估的变量。