Physiologic, ventilatory, and CNS transcriptomic response to chronic hypercapnia in goats

山羊对慢性高碳酸血症的生理、通气和中枢神经系统转录组反应

• 批准号: 10462046
• 负责人: Kirstyn Buchholz
• 金额: $ 3.82万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10462046
• 关键词: Acute; Adult; Affect; Air; Ambulatory Care Facilities; Animal Model; Animals; Bioinformatics; Biological Assay; Brain; Brain Stem; Breathing; Carbon Dioxide; Cell Nucleus; Chronic; Chronic Obstructive Pulmonary Disease; Data; Disease; Disease Progression; Dose; Exposure to; Financial compensation; Functional disorder; Gene Expression; Genes; Genome; Goals; Goat; Human Characteristics; Hypercapnia; Individual; Life; Lung; Lung diseases; Measures; Mechanics; Mediating; Modeling; Molecular; Molecular Biology Techniques; Neuronal Plasticity; Pathologic; Pathway Analysis; Pathway interactions; Patients; Persons; Phenotype; Physiological; Physiology; Predisposition; Preparation; Punch Biopsy; RNA; Rattus; Resistance; Respiratory Disease; Respiratory Failure; Risk Factors; Severities; Signal Pathway; Sinus; Stimulus; Symptoms; System; Techniques; Testing; Time; Tissue Extracts; Tissues; Training; blood pressure elevation; cDNA Library; cell type; cognitive function; cognitive system; comorbidity; differential expression; experience; experimental study; gene network; genome-wide; insight; mRNA sequencing; neurophysiology; novel; respiratory; response; transcriptome sequencing; transcriptomics

PROJECT SUMMARY Chronic obstructive pulmonary disease (COPD) affects 24 million US adults and over 1 billion people worldwide. COPD patients often present with chronic hypercapnia (CH, elevated CO2 in the body), with severity of CH increasing as the disease progresses. COPD is also marked by acute “attacks”, known as acute-on-chronic exacerbations, further increasing the level of hypercapnia within the body. Some COPD patients demonstrate tolerable, adaptive responses to the exacerbations, whereas others are prone to life-threatening severe hypercapnia and potentially fatal cardiorespiratory dysfunction. The severity of CH may be an important factor in setting the threshold of susceptibility to adaptive versus pathophysiologic responses. Indeed, our previous findings have shown goats chronically exposed to mild CH (6% inspired CO2, InCO2) demonstrated adaptations across multiple physiologic systems and tolerated further acute increases in InCO2 to 7 and 8% CO2. In contrast, in a preliminary study on one goat, chronically increasing InCO2 from mild to moderate levels (8% InCO2), resulted in pathophysiologic responses and the inability to compensate for acute further increases in InCO2 to 9% and 10%. The molecular underpinnings governing the protective/adaptive responses to chronic mild CH or pathophysiologic/maladaptive responses to chronic moderate CH are unclear. Accordingly, the overall goal of this proposal is to gain insight into CH-dependent molecular mechanisms which potentially underly both the protective/adaptive (AIM 1) and pathophysiologic/maladaptive physiologic (AIM 2) responses to increased CO2 challenges. For AIM 1 I will expose adult goats to 14 days (d) of room air (Group 1) or mild CH (Group 2). For AIM 2, I will expose goats to 7d of mild CH (6% CO2) followed by 7d of moderate CH (8% CO2) (Group 3). I will measure physiological/pathophysiologic responses across multiple physiologic systems during steady-state conditions and during acute CO2 challenges. Goats will be euthanized after these studies and I will extract tissue from key brainstem cardiorespiratory nuclei and utilize bulk-tissue mRNA sequencing (btRNA-Seq) to identify differentially expressed genes induced by both mild CH and moderate CH. I will use Ingenuity Pathway Analysis to identify canonical pathways and functional gene networks that are significantly altered during mild CH (6% CO2) alone, or 7d mild CH followed by 7d of moderate CH (8% CO2). Combining broad, unbiased techniques (btRNA-Seq) and physiologic studies will provide novel insights into the molecular mechanisms regulating cardiorespiratory control networks during CH. Further, results will yield information that will be critical in understanding the effect CH has on the physiologic dysfunction observed in outpatient clinics and ICUs. The training plan outlined in this proposal will provide me with the expertise needed to conduct physiologic experiments, molecular biology techniques, and bioinformatic analyses required for the proposed studies.

项目摘要 慢性阻塞性肺疾病（COPD）影响了2400万美国成年人，全球超过10亿人。 COPD患者经常出现慢性高碳酸盐（CH，体内二氧化碳升高），CH严重程度 随着疾病的进展而增加。 COPD还以急性“攻击”为标志 加剧，进一步增加了体内的高碳酸脂蛋白水平。一些COPD患者证明 对恶化的可忍受，适应性的反应，而其他人则容易威胁生命 高碳酸盐和潜在的致命性心肺功能障碍。 CH的严重程度可能是一个重要因素 在设定适应性与病理生理反应的易感性的阈值时。的确，我们以前的 调查结果已显示出长期暴露于轻度CH（6％启发的CO2，INCO2）的山羊展示了适应 在多个生理系统中，INC2至7％和8％CO2的耐受性急性增加。相比之下， 在一项对一只山羊的初步研究中，长期从轻度到中等水平（8％INCO2）逐渐增加了INCO2， 导致病理生理反应和无法补偿急性进一步增加INCO2的急性 9％和10％。管理受保护/适应性反应的分子基础对慢性轻度CH或 对慢性现代CH的病理生理/不良适应反应尚不清楚。彼此之间，总体目标 该提议是为了深入了解CH依赖性分子机制，这可能是 保护性/适应性（AIM 1）和病理生理/适应性生理学（AIM 2）对CO2增加的反应 挑战。对于目标1，我将使成年山羊暴露于14天（d）的房间空气（第1组）或轻度CH（第2组）。为了 AIM 2，我将使山羊暴露于7d的温和CH（6％CO2），然后将中度CH（8％CO2）（第3组）暴露于7天。我会 测量稳态期间多个生理系统的生理/病理生理反应 条件和急性二氧化碳挑战。这些研究后，山羊将被安乐死，我将提取组织 从关键的脑干心肺核并利用散装组织mRNA测序（BTRNA-SEQ）识别 由轻度CH和中度CH诱导的差异表达基因。我将使用Ingenuity途径分析 确定在轻度CH期间显着改变的规范途径和功能基因网络（6％ 二氧化碳）单独或7D轻度CH，然后是7d中度CH（8％CO2）。结合广泛的公正技术 （BTRNA-SEQ）和生理研究将提供对调节的分子机制的新见解 CH期间的心肺控制网络。此外，结果将产生至关重要的信息 了解CH对门诊诊所和ICU中观察到的生理功能障碍的影响。 该建议中概述的培训计划将为我提供进行生理所需的专业知识 拟议研究所需的实验，分子生物学技术和生物信息学分析。