MicroRNAs as Biomarkers for Obstructive Sleep Apnea

摘要 确定睡眠相关疾病严重程度的生物标志物和治疗的有效性， 最近被列为国家2021年睡眠研究计划中的头号关键机会。 美国国家睡眠障碍研究中心（National Center on Sleep Disorders Research，NCSDR）阻塞性睡眠呼吸暂停 (OSA)是常见的，有有限的生物标志物用于识别和管理的条件。具体使用 OSA生物标志物的病例包括：（i）改善病例识别，（ii）监测治疗的功效，和（iii） 提供关于谁将获得特定后果或个人如何应对的预后价值。 持续气道正压通气（CPAP）治疗。虽然可以使用不同的方法来定义 生物标志物，该项目将集中在microRNA，这是最近被证明是有前途的 OSA中的生物标志物。MicroRNA是小的非编码RNA，其改变蛋白质编码RNA的翻译。 它们的表达是动态的，并会受到许多挑战的影响，如缺氧。所有microRNA在 可以通过对所有短RNA进行测序来评估血液。先前的研究，尽管样本量小，表明 OSA病例和对照组之间microRNA表达的差异以及 表达可以识别患有OSA的个体，其将对CPAP治疗显示更大的血压反应。 使用互补测序方法和临床可行的定量PCR（qPCR），我们建议 验证并扩展这些初步观察结果。首先，我们将通过以下方法寻找OSA的特异性生物标志物： 评估年龄匹配的OSA病例和无OSA对照组之间microRNA谱的差异， 性别和体重指数，并且没有其他可能独立影响microRNA的基础条件 表情虽然鉴定OSA特异性microRNA很重要，但确定microRNA也很有用。 用于改进OSA病例识别，超出已知的临床风险因素。因此，该项目将招收一名 具有最低排除标准的较大病例对照样本，用于评估差异的预测值 在microRNA表达中。为了了解microRNA作为治疗相关生物标志物的效用， 将在CPAP治疗前和治疗后6个月对OSA进行研究。我们预计，一些特定于OSA的microRNA 将通过CPAP治疗正常化，从而提供有效性的客观测量。在所有OSA病例中，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。 来预测CPAP对血压的影响我们将对所有生物标志物进行稳健验证。首先利用 在发现分析中测序的相同样品，我们将通过临床可行的方法进行分析验证。 qPCR技术。microRNA签名的独立验证将在新样本中完成，包括病例 01项目的控制此外，微RNA和其他在生物学领域开发的特征的相对效用 计划，如多基因风险评分，将进行评估。总的来说，该项目直接符合所述 NCSDR的研究目标和本方案项目的具体目标。

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.