Table_1_Evaluation of the Association Between Genetic Variants in Circadian Rhythm Genes and Posttraumatic Stress Symptoms Identifies a Potential Functional Allele in the Transcription Factor TEF.DOCX

Previous studies suggest that genetic variants within genes affecting the circadian rhythm influence the development of posttraumatic stress symptoms (PTSS). In the present study, we used data from three emergency care-based cohorts to search genetic variants in circadian pathway genes previously associated with neuropsychiatric disorders for variants that influence PTSS severity. The three cohorts used included a discovery cohort of African American men and women enrolled following motor vehicle collision (n = 907) and two replication cohorts: one of multi-ethnic women enrolled following sexual assault (n = 274) and one of multi-ethnic men and women enrolled following major thermal burn injury (n = 68). DNA and RNA were collected from trauma survivors at the time of initial assessment. Validated questionnaires were used to assess peritraumatic distress severity and to assess PTSS severity 6 weeks, 6 months, and 1 year following trauma exposure. Thirty-one genetic variants from circadian rhythm genes were selected for analyses, and main effect and potential gene*stress and gene*sex interactions were evaluated. Secondary analyses assessed whether associated genetic variants affected mRNA expression levels. We found that six genetic variants across five circadian rhythm-associated genes predicted PTSS outcomes following motor vehicle collision (p < 0.05), but only two of these variants survived adjustment for multiple comparisons (False Discovery Rate < 5%). The strongest of these associations, an interaction between the PAR-zip transcription factor, thyrotroph embryonic factor (TEF) variant rs5758324 and peritraumatic distress, predicted PTSS development in all three cohorts. Further analysis of genetic variants in the genetic region surrounding TEFrs5758324 (±125,000 nucleotides) indicated that this allele showed the strongest association. Further, TEF RNA expression levels (determined via RNA-seq) were positively associated with PTSS severity in distressed individuals with at least one copy of the TEFrs5758324 minor allele. These results suggest that rs5758324 genetic variant in TEF, a regulator of clock-controlled genes and key mediator of the core circadian rhythm, influence PTSS severity in a stress-dependent manner.

既往研究表明，影响昼夜节律（circadian rhythm）的基因内遗传变异会影响创伤后应激症状（posttraumatic stress symptoms, PTSS）的发生发展。本研究依托3项基于急诊诊疗的队列数据，针对既往与神经精神疾病相关的昼夜节律通路基因中的遗传变异，开展了对PTSS严重程度具有影响的变异位点筛查。
本次纳入的3项队列包括：1项非洲裔美国男性与女性的发现队列（纳入于机动车碰撞事故后，n=907），以及2项重复验证队列：1项为性侵犯后纳入的多民族女性队列（n=274），另1项为重大热烧伤后纳入的多民族男性与女性队列（n=68）。研究人员在创伤幸存者初次评估时采集了其DNA与RNA样本，采用经过验证的问卷分别评估创伤即时痛苦严重程度，并在创伤暴露后6周、6个月及1年时评估PTSS严重程度。
本研究共选取来自昼夜节律相关基因的31个遗传变异开展分析，并评估了主效应以及潜在的基因×应激、基因×性别交互作用。次要分析则评估了相关遗传变异是否会影响信使RNA（messenger RNA, mRNA）的表达水平。
研究结果显示，在机动车碰撞事故队列中，来自5个昼夜节律相关基因的6个遗传变异可预测PTSS结局（p<0.05），但其中仅2个变异在校正多重比较后仍具有统计学意义（错误发现率<5%）。其中关联性最强的为PAR家族拉链转录因子促甲状腺胚胎因子（thyrotroph embryonic factor, TEF）的rs5758324变异与创伤即时痛苦的交互作用，该交互作用可在全部3项队列中预测PTSS的发生。
对TEF rs5758324所在遗传区域（±125000个核苷酸）内的遗传变异进行的进一步分析表明，该等位基因展现出最强的关联性。此外，在携带至少1份TEF rs5758324次要等位基因的痛苦个体中，通过RNA测序（RNA-seq）测得的TEF RNA表达水平与PTSS严重程度呈正相关。
上述结果表明，TEF基因——作为时钟调控基因的调控因子与核心昼夜节律的关键介导因子——内的rs5758324遗传变异可通过应激依赖的方式影响PTSS严重程度。