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.

每年数亿名患者暴露于全身麻醉药（GAS），从而产生可遗传的效果 天然气的公共卫生问题至关重要。需要调查气体的可遗传影响和 开发预防疗法也是由前所未有的上升表明，特别是在工业化国家， 神经发育障碍。这些疾病大多数的起源是未知的，许多疾病更常见 在男性中。为了构成有关该主题的临床研究的基础，拟议的临床前项目将测试以下 关于与气体的可遗传作用有关的机制，热工具和生物标志物的假设： 1）GA诱导的立体胁迫同源性皮质酮（CORT）的分泌对于主动性至关重要 亲本生殖细胞的表观遗传变化（F0生成），并且随着后代的延伸异常（F1） 一代）; 2）两种GABA能GA诱导的K+-2Cl-（KCC2）Cl- Exporter的损害，导致 抑制性GABAAR信号的损害以及GA诱导的F0需要增加Cort分泌的增加 神经行为缺陷； 3）F1雄性更脆弱，因为F0气体通过修改男性 - 特定的F1脑男性化。目标1：确定KCC2和Cort在主动性中的作用 Sevoflurane（Sevo）的代际作用。四种临床使用的气体具有部分重叠的机制 作用，塞沃，提案，氯胺酮和脱氧酸酯将被用作确定角色的药物工具 KCC2和Cort的F0大鼠将在产后日（P）56，P58和P60和伴侣暴露于气体 P85生成后代。 F0和F1大鼠将在高高的迷宫中评估 惊吓*，莫里斯水迷宫，恐惧激动的惊吓和强迫游泳测试以及评估休息和 应力诱导的下丘脑 - 垂体 - 肾上腺（HPA）轴活动*。 F0和F1中的全基因组DNA甲基化 F0和F1海马和外周血单核细胞中的生殖细胞和海马和RNA-SEQ将使用 为了深入了解表观基因质和转录组机制（* =将作为潜在的生物标志物评估 未来的人类研究）。目标2：确定较短的父母塞沃暴露是否不足以诱导 F0神经行为效应可以诱导F1效应，并确定cl-转运蛋白和糖皮质激素的作用 受体（GRS）。假设：较短的F0 SEVO暴露足以更新F0 HPA轴和 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 治疗作用。目标3：确定父母雄性估计效应的机制 暴露于Sevo。假设：F1雄性更脆弱，因为F0 Sevo会影响睾丸激素（T） - 调节大脑男性化。因为T通过雌激素受体α（ERα）和 通过T激活的雄激素受体（ARS），我们假设F1ERα - / - （但不是ERβ-/ - ）男性和/或F1野生 出生时用AR拮抗剂治疗的型男性会受到较小的影响。 KCC2增强或NKCC1 GR抑制作用将减轻F0 SEVO诱导的脑男性化的F1男性机制的变化。