Sulforaphane-mediated epigenetic modulation of ethanol-induced apoptosis and teratogenesis

产前接触乙醇是导致智力低下的主要原因。越来越多的证据表明 过度细胞死亡是乙醇引起的出生缺陷发病机制的主要组成部分。然而， 对于乙醇如何导致胚胎细胞凋亡的理解存在根本性的差距。 萝卜硫素 (SFN) 是西兰花芽中富含的一种化学物质。令人信服的证据表明 SFN 或富含 SFN 的西兰花芽通过激活 Nrf2 信号触发抗氧化酶的诱导 并预防癌症和其他疾病。最近发现SFN可以调节基因表达 通过表观遗传机制。我们最近证明 SFN 可以显着减少乙醇- 诱导神经嵴细胞（NCC）凋亡。我们的长期目标是发展 对抗乙醇致畸的有效策略；基于预防乙醇诱发的策略 通过靶向参与细胞凋亡的特定途径来实现细胞凋亡。本次专项行动的总体目标 该提案旨在阐明 SFN 表观遗传调控抗凋亡的表观遗传机制 基因表达和预防乙醇诱导的细胞凋亡，并开发安全有效的治疗方案 使用 SFN（一种安全的“营养药物”）降低 FASD 的发病率和严重程度。这个假设的中心假设 项目是乙醇诱导的表观遗传改变导致抗凋亡基因的异常表达， 导致胚胎过度凋亡和畸形，这可以通过表观遗传来预防 由 SFN 或富含 SFN 的西兰花芽提取物引起的调节。我们的假设是基于 以我们实验室产生的强有力的初步数据为基础。为了检验我们的假设，有以下具体目标 将解决： 目标 1：鉴定乙醇和乙醇在表观遗传上沉默的抗凋亡基因 在暴露于乙醇的 NCC 和小鼠胚胎中，SNF 可以解除抑制。目标 2：阐明机制 SFN 表观遗传调节抗凋亡基因的表达和乙醇中的细胞凋亡 暴露的 NCC 和小鼠胚胎。目标 3：检验抗-表观遗传调节的假设 母体饮食中富含 SFN 的西兰花芽提取物 (BSE) 引起的凋亡基因代表了一种新的治疗方法 预防乙醇致畸的策略。拟议的工作具有创新性，因为它重点关注 一种新方法，即抗凋亡基因的表观遗传调节，以防止乙醇诱导的细胞凋亡 致畸。该申请中描述的理论概念也具有高度创新性，因为这是 第一项研究试图通过使用从蔬菜中提取的生物活性化合物来预防胎儿酒精谱系障碍（FASD）。的 这项研究的结果将具有重要意义，因为它们有望说明实际应用的潜力 基于营养药物的 FASD 治疗策略。

Prenatal ethanol exposure is the leading known cause of mental retardation. Growing evidence suggests that excessive cell death is a major component of the pathogenesis of ethanol-induced birth defects. However, there is a fundamental gap in understanding how ethanol leads to apoptotic cell death in embryos. Sulforaphane (SFN) is a chemical that is abundant in broccoli sprouts. Compelling evidence indicates that SFN or SFN-rich broccoli sprouts trigger the induction of antioxidant enzymes through activation of Nrf2 signaling and prevent cancer and other diseases. It was recently discovered that SFN can regulate gene expression through epigenetic mechanisms. We have recently demonstrated that SFN can significantly diminish ethanol- induced apoptosis in neural crest cells (NCCs). Our long-term goal is directed toward the development of effective strategies against ethanol's teratogenesis; strategies based on prevention of ethanol-induced apoptosis through targeting specific pathways involved in apoptosis. The overall objective of this particular proposal is to elucidate the epigenetic mechanisms by which SFN epigenetically regulates the anti-apoptotic gene expression and prevent ethanol-induced apoptosis, and to develop a safe and effective regimen to diminish the incidence and severity of FASD using SFN, a safe “nutraceutical”. The central hypothesis of this project is that ethanol-induced epigenetic alterations lead to aberrant expression of the anti-apoptotic genes, resulting in excessive apoptosis and malformations in embryos, which can be prevented by epigenetic modulation elicited by SFN or SFN-rich broccoli sprout extract. Our hypothesis has been formulated on the basis of strong preliminary data produced in our laboratory. To test our hypothesis, the following specific aims will be addressed: Aim 1: To identify the anti-apoptotic genes that are epigenetically silenced by ethanol and derepressed by SNF in ethanol-exposed NCCs and mouse embryos. Aim 2: To elucidate the mechanisms by which SFN epigenetically modulates the expression of the anti-apoptotic genes and apoptosis in ethanol- exposed NCCs and mouse embryos. Aim 3: To test the hypothesis that epigenetic modulation of the anti- apoptotic genes by maternal dietary SFN-rich broccoli sprout extract (BSE) represents a novel therapeutic strategy for preventing ethanol-induced teratogenesis. The proposed work is innovative, because it focuses on a novel approach, epigenetic modulation of the anti-apoptotic genes, to preventing ethanol-induced teratogenesis. The theoretical concept described in the application is also highly innovative because this is the first study attempting to prevent FASD through the use of bioactive compounds derived from a vegetable. The results from this study will be significant, because they are expected to illustrate the potential of a practical nutraceutical-based therapeutic strategy for FASD.