Role of exosomes in the coordinated migration of neural crest cells and placodes and ethanol-induced teratogenesis

胎儿酒精谱系障碍（FASD）是智力迟钝的主要已知原因。越来越多的证据 表明神经嵴细胞（NCC）和基板（PC）迁移的破坏对乙醇的主要贡献- 导致颅面和颅神经缺损。然而，在理解如何做到这一点方面存在着根本性的差距。 乙醇破坏了胚胎中NCC和PC的协调迁移。外来体最近出现， 一种重要的细胞间通讯方式。外泌体携带并转移多种活性分子， 包括microRNA，作用于局部和远处的受体细胞，并影响它们的功能。我们最近 证明来源于人NCC的外来体介导乙醇诱导的SDF 1/CXCR 4抑制 信号传导，随后破坏了NCC和PC的协调迁移， 和PC迁移有助于乙醇诱导的斑马鱼胚胎颅面和脑神经缺陷。 该提案的总体目标是阐明外泌体介导乙醇的机制， 诱导抑制SDF 1/CXCR 4信号传导，破坏NCC和PC的协调迁移， 颅面和颅神经缺陷，并建立植物来源的外泌体样纳米颗粒作为一种可行的 预防FASD的策略。该项目的中心假设是，乙醇破坏了协调的 通过NCC-derived exosome介导的SDF 1/CXCR 4信号转导抑制NCCs和PC的迁移， 随后导致颅面和颅神经缺陷，以及SDF 1/CXCR 4信号传导调节 葡萄外泌体样纳米颗粒（GELN）或装载有miRNA调节剂的GELN可以防止乙醇- 诱导致畸为了检验我们的假设，将讨论以下具体目标：目标1： 表征外泌体在乙醇诱导的NCC和PC协调迁移的破坏中的作用 在体外和斑马鱼胚胎中。目的2：阐明NCC来源的外泌体介导的细胞内信号转导的机制。 乙醇诱导的SDF 1/CXCR 4信号传导的抑制和NCC协调迁移的破坏， PCs.目的3：检验来源于NCC的外来体介导乙醇诱导的颅面神经损伤的假设， 脑神经缺损与GELN或加载有miRNA的GELN对SDF 1/CXCR 4信号传导的调节 FASD调节剂代表了预防FASD的新治疗策略。拟议的工作是创新的， 因为这是第一项试图通过新认识到的外泌体在预防FASD中的作用的研究。 SDF 1/CXCR 4信号通路在协调迁移调控中的作用 NCC和PC。这也是首次尝试使用可食用植物来源的类外泌体来预防FASD。 纳米粒子这项研究有望描述一种新的外泌体介导的细胞间 NCCs和PCs在乙醇诱导的致畸作用中的相互作用。这样的结果将是显著的， 因为从这项研究中获得的见解将有助于阐明FASD和产量的潜在机制 基于外泌体的预防FASD的策略。

Fetal Alcohol Spectrum Disorders (FASD) is the leading known cause of mental retardation. Growing evidence suggests a major contribution of the disruption of neural crest cell (NCC) and placode (PC) migration to ethanol- induced craniofacial and cranial nerve defects. However, there is a fundamental gap in understanding how ethanol disrupts the coordinated migration of NCCs and PCs in embryos. Exosomes have recently emerged as an important mode of intercellular communication. Exosomes carry and transfer a variety of active molecules, including microRNAs to and act on local and distant recipient cells, and affect their functions. We have recently demonstrated that exosomes derived from human NCCs mediated ethanol-induced repression of SDF1/CXCR4 signaling and subsequently disrupted the coordinated migration of NCCs and PCs and that disruption of NCC and PC migration contributes to ethanol-induced craniofacial and cranial nerve defects in zebrafish embryos. The overall objective of this proposal is to elucidate the mechanisms by which exosomes mediate ethanol- induced repression of SDF1/CXCR4 signaling, disruption of the coordinated migration of NCCs and PCs, and craniofacial and cranial nerve defects, and to establish plant-derived exosome-like nanoparticles as a feasible strategy for the prevention of FASD. The central hypothesis of this project is that ethanol disrupts the coordinated migration of NCCs and PCs through NCC-derived exosome-mediated repression of SDF1/CXCR4 signaling, subsequently leading to craniofacial and cranial nerve defects, and that modulation of SDF1/CXCR4 signaling by grape exosome-like nanoparticles (GELNs) or GELNs loaded with miRNA modulators can prevent ethanol- induced teratogenesis. To test our hypothesis, the following specific aims will be addressed: Aim 1: To characterize the role of exosomes in ethanol-induced disruption of the coordinated migration of NCCs and PCs in vitro and in zebrafish embryos. Aim 2: To elucidate the mechanisms by which NCC-derived exosomes mediate ethanol-induced repression of SDF1/CXCR4 signaling and disruption of the coordinated migration of NCCs and PCs. Aim 3: To test the hypothesis that exosomes derived from NCCs mediate ethanol-induced craniofacial and cranial nerve defects and that modulation of SDF1/CXCR4 signaling by GELNs or GELNs loaded with miRNA modulators represents a novel therapeutic strategy for preventing FASD. The proposed work is innovative, because this is the first study attempting to prevent FASD through the newly recognized role of exosomes in intercellular communication and the actions of SDF1/CXCR4 signaling in modulating the coordinated migration of NCCs and PCs. It also represents the first attempt to prevent FASD using edible plant-derived exosome-like nanoparticles. The proposed research is expected to characterize a novel, exosome-mediated intercellular interaction between NCCs and PCs during ethanol-induced teratogenesis. Such results will be significant, because the insights gained from this study will help in elucidating the mechanisms underlying FASD and yield exosome-based strategies for the prevention of FASD.