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

产前酒精暴露是导致智力迟钝的主要已知原因。越来越多的证据表明， 过度的细胞死亡是乙醇诱导的出生缺陷的发病机理的主要组成部分。然而，在这方面， 在理解乙醇如何导致胚胎中的凋亡性细胞死亡方面存在根本性的差距。 萝卜硫素（SFN）是一种在西兰花芽中含量丰富的化学物质。令人信服的证据表明， 或SFN丰富的西兰花芽通过激活Nrf 2信号传导触发抗氧化酶的诱导 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。我们最近证明了SFN可以显著减少乙醇- 诱导神经嵴细胞凋亡。我们的长期目标是发展 预防乙醇致畸作用的有效策略 通过靶向参与细胞凋亡的特定途径来诱导细胞凋亡。这一特别行动的总体目标 建议是阐明SFN表观遗传调节抗凋亡的表观遗传机制， 基因表达和防止乙醇诱导的细胞凋亡，并开发一种安全有效的方案， 使用安全的“营养药物”SFN降低FASD的发病率和严重程度。这个问题的核心假设是 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学来预防。 通过SFN或富含SFN的西兰花芽提取物引起的调节。我们的假设是建立在 这是我们实验室初步数据的基础。为了验证我们的假设，以下具体目标 目的1：鉴定乙醇引起表观遗传沉默的抗凋亡基因， SNF在乙醇暴露的NCC和小鼠胚胎中的去抑制作用。目的2：阐明机制， SFN在表观遗传学上调节乙醇中抗凋亡基因的表达和凋亡， 暴露的NCC和小鼠胚胎。目的3：为了检验抗-HCV基因的表观遗传调节的假说， 母亲饮食中富含SFN的西兰花芽提取物（BSE）的凋亡基因代表了一种新的治疗方法， 预防乙醇诱导的致畸作用的策略。拟议的工作是创新的，因为它侧重于 一种新的方法，抗凋亡基因的表观遗传调节，以防止乙醇诱导的 致畸作用申请中描述的理论概念也是高度创新的，因为这是 这是第一项尝试通过使用植物中的生物活性化合物来预防FASD的研究。的 这项研究的结果将是重要的，因为他们预计将说明一个实际的 FASD的营养保健品为基础的治疗策略。

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.