Mechanisms and blood-based biomarkers of intergenerational neurobehavioral effects of general anesthetics

全身麻醉药代际神经行为效应的机制和血液生物标志物

• 批准号: 10538703
• 负责人: ANATOLY E MARTYNYUK
• 金额: $ 47.8万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-20 至 2027-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10538703
• 关键词: ARHGEF5 gene; Acoustics; Acute; Affect; Alcohols; Androgen Receptor; Attention; Attention deficit hyperactivity disorder; Behavior; Behavioral; Biological Markers; Birth; Brain; Bumetanide; Clinical; Clinical Research; Corticosterone; DNA Methylation; Data; Defect; Developed Countries; Disease; Endocrine Disruptors; Enhancers; Epigenetic Process; Estradiol; Estrogen Receptor alpha; Etomidate; Exposure to; Flutamide; Foundations; Fright; Future; Gene Expression; General anesthetic drugs; Generations; Genes; Germ Cells; Glucocorticoid Receptor; Heritability; Hippocampus (Brain); Hormones; Human; Impairment; Ketamine; Link; Masculine; Measures; Mediating; Mifepristone; Modification; Neurodevelopmental Disorder; Neurosecretory Systems; Operative Surgical Procedures; Parents; Partner in relationship; Patients; Pharmacology; Pilot Projects; Prevention strategy; Preventive therapy; Procedures; Process; Propofol; Public Health; Rattus; Receptor Inhibition; Research; Rest; Rodent; Role; Signal Transduction; Sprague-Dawley Rats; Steroids; Stress; Testing; Testis; Testosterone; Therapeutic; Therapeutic Effect; Work; antagonist; autism spectrum disorder; base; bisulfite sequencing; blood-based biomarker; design; developmental disease; differential expression; environmental stressor; epigenome; epigenomics; experimental study; follow-up; forced swim test; genome-wide; genome-wide analysis; hypothalamic-pituitary-adrenal axis; inhibitor; insight; intergenerational; male; monocyte; morris water maze; neurobehavioral; offspring; perinatal brain; peripheral blood; postnatal; potential biomarker; pre-clinical; prepulse inhibition; resilience; sevoflurane; sperm cell; stressor; tool; transcriptome sequencing; transcriptomics; transmission process; young adult

Hundreds of millions of patients are exposed to general anesthetics (GAs) each year, making the heritable effects of GAs a public health issue of paramount importance. The need to investigate the heritable effects of GAs and develop preventative therapies is also indicated by an unprecedented rise, particularly in industrialized countries, of neurodevelopmental disorders. The origin of most of these disorders is unknown and many are more common in males. To form the basis for clinical studies on this topic, the proposed preclinical project will test the following hypotheses regarding the mechanisms, therapeutic tools, and biomarkers pertaining to heritable effects of GAs: 1) GA-induced secretion of the steroid stress hormone corticosterone (CORT) is essential in the initiation of epigenetic changes in parental germ cells (F0 generation) and, by extension, abnormalities in offspring (F1 generation); 2) both GABAergic GA-induced impairment of the K+-2Cl- (KCC2) Cl- exporter, resulting in impairment of inhibitory GABAAR signaling, and an increase in CORT secretion are required for GA-induced F0 neurobehavioral defects; and 3) F1 males are more vulnerable because F0 GAs act via modification of male- specific F1 brain masculinization. Aim 1: Determine the roles of KCC2 and CORT in the initiation of intergenerational effects of sevoflurane (SEVO). Four clinically used GAs with partially overlapping mechanisms of action, SEVO, propofol, ketamine and etomidate, will be used as pharmacological tools to determine the roles of KCC2 and CORT. The F0 rats will be exposed to GAs on postnatal day (P) 56, P58, and P60 and mated on P85 to generate offspring. The F0 and F1 rats will be evaluated in the elevated plus maze, prepulse inhibition of startle*, Morris water maze, fear-potentiated startle, and forced swimming test and by assessing resting and stress-induced hypothalamic-pituitary-adrenal (HPA) axis activity*. Genome-wide DNA methylation in F0 and F1 germ cells and hippocampi and RNA-seq in F0 and F1 hippocampi and peripheral blood monocytes* will be used to gain insight into epigenomic and transcriptomic mechanisms (* = will be evaluated as potential biomarkers for future human studies). Aim 2: Determine whether shorter parental SEVO exposure that is not sufficient to induce F0 neurobehavioral effects can induce F1 effects, and determine the roles of Cl- transporters and glucocorticoid receptors (GRs). Hypotheses: Shorter F0 SEVO exposure is sufficient to upregulate the F0 HPA axis and reprogram the germline, inducing F1 defects. KCC2 enhancement or Na+-K+-Cl- (NKCC1) and GR inhibition have therapeutic effects. Aim 3: Determine the mechanisms of male-biased intergenerational effects of parental exposure to SEVO. Hypotheses: F1 males are more vulnerable because F0 SEVO affects testosterone (T)- regulated brain masculinization. Because T acts through 17β-estradiol (E2) at estrogen receptor α (ERα) and via T-activated androgen receptors (ARs), we hypothesize that F1 ERα-/- (but not ERβ-/-) males and/or F1 wild type males treated with the AR antagonist flutamide at birth will be less affected. KCC2 enhancement or NKCC1 and GR inhibition will alleviate the F0 SEVO-induced changes in F1 male mechanisms of brain masculinization.

每年有数亿患者接触全身麻醉剂 (GA)，从而产生遗传效应 GA 是一个至关重要的公共卫生问题。需要研究 GA 的遗传效应和 开发预防性疗法的空前增长也表明，特别是在工业化国家， 神经发育障碍。大多数这些疾病的起源尚不清楚，而且许多疾病更为常见 在男性中。为了形成该主题临床研究的基础，拟议的临床前项目将测试以下内容 关于 GA 遗传效应的机制、治疗工具和生物标志物的假设： 1) GA 诱导的类固醇应激激素皮质酮 (CORT) 的分泌对于启动 亲代生殖细胞（F0 代）的表观遗传变化，以及后代（F1 代）的异常 一代）; 2) GABAergic GA 诱导的 K+-2Cl- (KCC2) Cl- 输出蛋白损伤，导致 GA 诱导的 F0 需要抑制性 GABAAR 信号传导受损和 CORT 分泌增加 神经行为缺陷； 3) F1 雄性更容易受到伤害，因为 F0 GA 通过修饰雄性来发挥作用 特定的 F1 大脑男性化。目标 1：确定 KCC2 和 CORT 在启动中的作用 七氟烷（SEVO）的代际效应。四种临床使用的具有部分重叠机制的 GA SEVO、异丙酚、氯胺酮和依托咪酯将用作药理学工具来确定其作用 KCC2 和 CORT。 F0 大鼠将在出生后第 56 天 (P)、P58 和 P60 接触 GA，并于 P85产生后代。 F0和F1大鼠将在高架十字迷宫中进行评估，预脉冲抑制 惊吓*、莫里斯水迷宫、恐惧增强惊吓和强迫游泳测试，并通过评估休息和 压力诱发的下丘脑-垂体-肾上腺 (HPA) 轴活动*。 F0 和 F1 中的全基因组 DNA 甲基化 将使用生殖细胞和海马以及 F0 和 F1 海马和外周血单核细胞* 中的 RNA-seq 深入了解表观基因组和转录组机制（* = 将被评估为潜在的生物标志物 未来的人类研究）。目标 2：确定是否较短的父母 SEVO 暴露不足以诱导 F0神经行为效应可以诱导F1效应，并决定Cl-转运蛋白和糖皮质激素的作用 受体（GR）。假设：较短的 F0 SEVO 暴露足以上调 F0 HPA 轴， 重新编程种系，诱导 F1 缺陷。 KCC2 增强或 Na+-K+-Cl- (NKCC1) 和 GR 抑制具有 治疗效果。目标 3：确定父母亲代际影响中男性偏向的机制 接触 SEVO。假设：F1 雄性更容易受到伤害，因为 F0 SEVO 影响睾酮 (T)- 调节大脑男性化。因为 T 通过 17β-雌二醇 (E2) 作用于雌激素受体 α (ERα)， 通过 T 激活的雄激素受体 (AR)，我们假设 F1 ERα-/-（但不是 ERβ-/-）雄性和/或 F1 野生 出生时接受 AR 拮抗剂氟他胺治疗的男性受影响较小。 KCC2 增强或 NKCC1 GR抑制将减轻F0 SEVO诱导的F1雄性大脑男性化机制的变化。