MicroRNAs as Biomarkers for Obstructive Sleep Apnea

摘要 确定睡眠相关疾病的严重程度和治疗效果的生物标记物 最近被评为国家睡眠研究计划2021年的头号关键机遇 健康研究所和国家睡眠障碍研究中心(NCSDR)。而阻塞性睡眠呼吸暂停 阻塞性睡眠呼吸暂停综合征(OSA)很常见，但用于识别和管理这种疾病的生物标志物有限。特定用途 阻塞性睡眠呼吸暂停综合征生物标记物的案例包括：(I)改进病例识别，(Ii)监测治疗效果，以及(Iii) 提供关于谁将受到特定后果或个人如何应对的预测价值 持续气道正压(CPAP)治疗。虽然可以使用不同的方法来定义 生物标记物，这个项目将专注于microRNAs，这是最近被证明是有希望的 阻塞性睡眠呼吸暂停低氧血症的生物标志物。MicroRNAs是一种小的非编码RNA，可以改变蛋白质编码RNA的翻译。 它们的表达是动态的，并受到许多挑战的影响，如低氧。所有microRNA在大肠杆菌中的表达 血液可以通过对所有短RNA进行测序来进行评估。先前的研究，尽管样本量很小，但表明 阻塞性睡眠呼吸暂停患者与对照组的microRNA表达差异及microRNA差异 表达可以识别对CPAP治疗有较大血压反应的阻塞性睡眠呼吸暂停患者。 利用互补测序方法和临床可行的定量聚合酶链式反应(QPCR)，我们建议 验证并推广这些初步观察。首先，我们将通过以下方式寻找针对OSA的生物标记物 评估年龄匹配的阻塞性睡眠呼吸暂停患者和非阻塞性睡眠呼吸暂停患者之间的microRNA图谱的差异。 性别和体重指数，且没有其他可能独立影响microRNA的潜在条件 表情。虽然识别OSA特有的microRNAs很重要，但确定microRNAs也很有用 有助于改进OSA病例识别，超越已知的临床风险因素。因此，该项目将招收一名 具有最小排除标准以评估差异的预测价值的较大病例对照样本 在microRNA的表达中。为了了解microRNAs作为治疗相关生物标记物的用途， 在CPAP治疗前和治疗6个月后分别进行OSA检查。我们预计一些针对阻塞性睡眠呼吸暂停综合征的microRNA 将随着CPAP治疗而正常化，从而提供一个客观的有效性衡量标准。在所有OSA案例中，我们 将评估24小时动态血压，以验证和扩展最近关于microRNA签名的报告 这可以预测对CPAP的血压反应。我们将对所有生物标志物进行强有力的验证。首先，使用 在发现分析中测序的相同样本，我们将通过临床可行的方式进行分析验证 QPCR技术。将在新样本中对microRNA签名进行独立验证，包括病例 和来自Project 01的控件。此外，MicroRNAs和其他签名的相对效用在 计划，如多基因风险评分，将进行评估。总体而言，该项目直接与所述 NCSDR的研究目标和本计划项目的具体目标。

ABSTRACT Identification of biomarkers of the severity of sleep-related conditions and the effectiveness of therapies was recently presented as the number one critical opportunity in the 2021 Sleep Research Plan from the National Institutes of Health and National Center on Sleep Disorders Research (NCSDR). While obstructive sleep apnea (OSA) is common, there are limited biomarkers for identification and management of the condition. Specific use cases for an OSA biomarker include: (i) improving case identification, (ii) monitoring efficacy of therapy, and (iii) providing prognostic value with respect to who will get particular consequences or how individuals respond to continuous positive airway pressure (CPAP) treatment. While different approaches can be used to define biomarkers, this project will focus on microRNAs, which have very recently been shown to be promising biomarkers in OSA. MicroRNAs are small non-coding RNAs that alter the translation of protein coding RNA. Their expression is dynamic and altered by many challenges, such as hypoxia. Expression of all microRNAs in blood can be assessed by sequencing all short RNAs. Prior studies, albeit with small sample sizes, suggest differences in microRNA expression between OSA cases and controls and that differences in microRNA expression can identify individuals with OSA who will show larger blood pressure responses to CPAP treatment. Using complementary sequencing approaches and clinically-feasible quantitative PCR (qPCR), we propose to validate and extend these initial observations. First, we will seek biomarkers that are specific to OSA by evaluating differences in microRNA profiles between cases with OSA and controls without OSA matched for age, sex, and body mass index and without other underlying conditions that could independently affect microRNA expression. While identifying microRNAs specific to OSA is important, it is also useful to determine microRNAs useful for improving OSA case identification beyond known clinical risk factors. Thus, this project will enroll a larger case-control sample with minimal exclusion criteria in which to assess the predictive value of differences in microRNA expression. To understand the utility of microRNAs as treatment-related biomarkers, cases with OSA will be studied before and after 6 months of CPAP. We anticipate that some microRNAs specific to OSA will normalize with CPAP treatment, thus providing an objective measure of effectiveness. In all OSA cases, we will assess 24-hour ambulatory blood pressure to validate and extend recent reports of a microRNA signature that predicts blood pressure response to CPAP. We will conduct robust validation for all biomarkers. First, using the same samples sequenced in discovery analyses, we will perform analytical validation via clinically-feasible qPCR techniques. Independent validation of microRNA signatures will be done in new samples, including cases and controls from Project 01. Moreover, the relative utility of microRNAs and other signatures developed in the Program, such as polygenic risk scores, will be assessed. Overall, this Project is directly aligned with the stated research objectives of the NCSDR and the specific goals of this Program Project.