Validating a minimally-invasive assay to study the genomic fingerprint of stress

描述（由申请人提供）：转化应激研究旨在阐明心理应激影响人类疾病病理生理的生物学机制。这一领域的进展一直受到技术差距的挑战，这迫使研究人员要么采用全面但有创的方法（例如，静脉穿刺后的基因组分析），要么采用微创但不全面的方法（例如，唾液测定）。的目标。我们的目标是验证一种全面的、微创的方法来研究人类的应激生物学，通过测试微创采血方法，即干燥血斑和毛细血管采血，是否可以用于检测全基因组表达模式的应激相关变化。方法。首先，为了实验室验证，我们将从10名健康志愿者的干血斑点、毛细血管血和静脉血中收集RNA。然后，我们将通过微创标本采集程序测试基因组图谱的可靠性、有效性和敏感性。这些测试将告知在临床验证中选择干血斑点或毛细管血作为标本采集。其次，为了临床验证，我们将要求1250对16岁的英国同卵双胞胎的人口代表性样本，通过双胞胎早期发展研究（TEDS）进行随访，完成计算机版的感知压力量表，我们将选择35对最不一致的双胞胎。然后，我们将测试微创微阵列分析是否可以识别具有相同DNA序列但不同压力暴露的年轻个体对的基因表达差异。的影响。研究压力基因组指纹的微创试验的验证将为研究人员提供在更大、更具人口代表性和更年轻的人类样本中进行压力生物学综合评估所需的技术。

DESCRIPTION (provided by applicant): Translational stress research has aimed to elucidate the biological mechanisms through which psychological stress influences disease pathophysiology in humans. Advancement in this field has been challenged by a technology gap that forces researchers to employ either comprehensive but invasive methods (e.g., genomic profiling after venipuncture) or minimally-invasive but non-comprehensive methods (e.g., salivary assays). AIM. We aim to validate a comprehensive and minimally-invasive method to study stress biology in humans, by testing whether minimally-invasive blood collection methods, namely dried blood spots and capillary blood collection, could be used to detect stress-related changes in genome-wide expression patterns. METHODS. First, for laboratory validation, we will collect RNA from dried blood spots, capillary blood, and venous blood in 10 healthy volunteers. We will then test the reliability, the validity, and the sensitivity of genomic profiles from minimally-invasive specimen-collection procedures. These tests will inform the choice of dried blood spots or capillary blood for specimen collection in a clinical validation. Second, for clinical validation, we will ask a population-representative sample of 1,250 pairs of 16-year-old UK monozygotic twins followed-up by the Twins Early Development Study (TEDS) to complete a computerized version of the Perceived Stress Scale and we will select the 35 most discordant twin pairs on this measure. We will then test whether microarray analysis from minimally-invasive assays can identify gene expression differences in pairs of young individuals with identical DNA sequences but different exposure to stress. IMPLICATIONS. The validation of a minimally-invasive assay to study the genomic fingerprint of stress will provide researchers with the technology needed to perform comprehensive assessment of stress biology in larger, more population-representative, and younger human samples.