Will therapeutic dosing of fatty acid amino hydrolase inhibitors disrupt neuronal

描述（由申请方提供）：这是一项响应PAR-08-216“发育药理学”的R21申请。减缓内源性大麻素anandamide降解的抑制剂在治疗焦虑，抑郁和疼痛方面表现出很大的希望，并且正在进行后两种适应症的临床试验。这些抑制剂通过阻断脂肪酸酰胺水解酶（FAAH）的水解来增加大麻素水平，从而主要通过CB 1大麻素受体增强内源性大麻素信号传导。FAAH抑制剂的急性副作用被很好地描述并且相对轻微。然而，FAAH抑制剂正在开发用于长期使用，这些药物的长期影响，特别是在大脑发育过程中的影响，研究得很少，也没有很好地了解。内源性大麻素如大麻素在整个身体的许多生理过程中起着关键作用。这些过程之一是神经发育，其中内源性大麻素调节神经发生、神经元迁移和轴突寻路。在神经发育过程中内源性大麻素信号的遗传或药理学破坏改变CNS发育。拟议的研究将使用小鼠和多学科方法来确定治疗活性剂量的FAAH抑制剂是否通过解决两个特定目标将花生四烯酸和相关N-酰基酰胺的胚胎脑水平提高到足以损害神经发育和随后的CNS功能（行为和神经元兴奋性）的水平：目标1。FAAH抑制剂的治疗剂量是否会抑制胚胎脑FAAH活性，导致异常神经发育？我们将使用质谱法来确定治疗剂量的FAAH抑制剂是否会增加发育中大脑中的大麻素和相关的N-酰基酰胺。然后我们将确定这些FAAH抑制剂是否影响神经发生、神经元迁移或轴突寻路。大麻素受体在这些过程中的参与将使用药理学或敲除方法进行评估。目标2.围产期FAAH抑制剂的治疗剂量是否会导致成年期行为和突触功能/可塑性的持续损害？小鼠将在围产期期间用在第一个特定目的中鉴定的FAAH抑制剂处理。然后，这些小鼠将以性别特异性的方式接受成年后的焦虑，药物偏好，空间学习和恐惧条件反射的行为测试。此外，我们将研究突触可塑性和兴奋/抑制平衡在成年期。通过完成这两个目标，我们将深入了解FAAH阻断对神经发育的影响，并将获得有关妊娠期间FAAH抑制的潜在后果的有价值的数据。 公共卫生相关性：内源性大麻素的分解抑制剂anandamide正在进行疼痛和抑郁症的临床试验。我们已经发现，内源性大麻素信号的干扰，包括抑制anandamide分解，导致神经发育紊乱。在拟议的工作中，我们将确定治疗剂量的花生四烯酸降解抑制剂是否会对神经发育产生不良影响。

DESCRIPTION (provided by applicant): This is an R21 application in response to PAR-08-216, "Developmental Pharmacology". Inhibitors that slow the degradation of the endocannabinoid anandamide show great promise for treating anxiety, depression, and pain, and are in clinical trials for the latter two indications. These inhibitors increase anandamide levels by blocking its hydrolysis by fatty acid amide hydrolase (FAAH) leading to enhanced endocannabinoid signaling, primarily through CB1 cannabinoid receptors. The acute side effects of FAAH inhibitors are well delineated and relatively mild. However, FAAH inhibitors are being developed for chronic use and the long-term effects of these drugs, particularly on the brain as it develops, are poorly studied and not well understood. Endocannabinoids such as anandamide play key roles in numerous physiological processes throughout the body. One of these processes is neurodevelopment where endocannabinoids modulate neurogenesis, neuronal migration, and axonal pathfinding. Genetic or pharmacological disruption of endocannabinoid signaling during neurodevelopment alters CNS development. The proposed studies will use mice and a multidisciplinary approach to determine if therapeutically active doses of FAAH inhibitors raise embryonic brain levels of anandamide and related N-acyl amides to levels sufficient to impair neurodevelopment and later CNS function (behavior and neuronal excitability) by addressing two specific aims: Aim 1. Will therapeutic dosing of FAAH inhibitors suppress embryonic brain FAAH activity leading to abnormal neurodevelopment? We will use mass spectrometry to determine if therapeutic doses of FAAH inhibitors increase anandamide and related N-acyl amides in developing brain. We will then determine if these FAAH inhibitors affect neurogenesis, neuronal migration, or axonal pathfinding. The involvement of cannabinoid receptors in these processes will be evaluated using pharmacological or knockout approaches. Aim 2. Will therapeutic dosing of FAAH inhibitors during the perinatal period lead to sustained impairment of behavior and synaptic function/plasticity during adulthood? Mice will be treated through the perinatal period with FAAH inhibitors identified in the first specific aim. These mice will then undergo behavioral testing as adults in a gender-specific fashion for anxiety, drug preference, spatial learning, and fear conditioning. In addition we will examine synaptic plasticity and excitation/inhibition balance during adulthood. By completing these two aims we will gain insight into the neurodevelopmental effects of FAAH blockade and will be provided with valuable data on the potential consequences of FAAH inhibition during pregnancy. PUBLIC HEALTH RELEVANCE: Inhibitors of the breakdown of the endogenous cannabinoid, anandamide, are undergoing clinical trials for pain and depression. We have found that perturbation of endocannabinoid signaling, including inhibition of anandamide breakdown, leads to derangements in neurodevelopment. In the proposed work we will determine if therapeutic doses of anandamide degradation inhibitors detrimentally affect neurodevelopment.