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)是目前已知的导致智力低下的主要原因。越来越多的证据 表明神经脊细胞(NCC)和胎盘(PC)迁移到乙醇的破坏是主要贡献- 所致的颅面神经和颅神经损伤。然而，在理解如何做到这一点上存在着根本的差距 乙醇破坏了NCC和PC在胚胎中的协调迁移。外显体最近已经出现为 细胞间通讯的一种重要方式。胞外体携带和转移各种活性分子， 包括去往和作用于本地和远程受体细胞的microRNA，并影响它们的功能。我们最近做了 人神经干细胞来源的外切体介导乙醇诱导的SDF1/CXCR4抑制 并随后扰乱了NCC和PC的协调迁移以及NCC的中断 在斑马鱼胚胎中，PC的迁移导致了乙醇引起的颅面神经和脑神经缺陷。 这项建议的总体目标是阐明外切体介导乙醇的机制。 诱导SDF1/CXCR4信号的抑制，破坏NCC和PC的协调迁移，以及 颅面和颅神经缺损，并建立植物来源的外体样纳米粒作为可行的 预防FASD的战略。该项目的中心假设是乙醇破坏了协调的 NCC来源的外切体介导的SDF1/CXCR4信号抑制NCC和PC的迁移， 从而导致颅面神经和脑神经缺陷，以及SDF1/CXCR4信号的调节 通过葡萄外切体样纳米颗粒(GELN)或负载miRNA调节剂的GELN可以阻止乙醇- 诱发畸形。为了验证我们的假设，我们将探讨以下具体目标：目标1： 表征外切体在乙醇诱导的NCC和PC协调迁移中断中的作用 在体外和斑马鱼胚胎中。目的2：阐明NCC来源的外切体介导的机制 乙醇诱导SDF1/CXCR4信号的抑制和NCC与CXCR4协同迁移的破坏 个人电脑。目的3：验证神经干细胞来源的外切体介导乙醇诱导的颅面神经损伤的假说。 GELN或负载miRNA的GELN对脑神经损伤及SDF1/CXCR4信号的调控 调节剂代表了预防FASD的一种新的治疗策略。建议的工作是创新的， 因为这是第一项试图通过新认识的外切体在 细胞间通讯及SDF1/CXCR4信号在协调迁移中的作用 NCC和PC。这也是首次尝试使用可食用植物来源的类似外切体来预防FASD 纳米粒子。这项拟议的研究有望描述一种新的、外切体介导的细胞间 酒精致畸过程中NCCs与PC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和收益率背后的机制 以外显子为基础的FASD预防策略。

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.