Investigating neurodevelopmental toxicity of perfluoroalkyl acids and their derivatives in human brain organoids models

摘要 全氟烷基和多氟烷基物质及其衍生物(PFAS)是一种工业化学品，具有广泛的 在环境中传播，包括野生动物和人类的血清。它们的渗透性和较长的半衰期 提出了关于它们的毒性的问题，特别是关于对发育中的胎儿的影响。几个 研究表明，这些化合物穿过胎盘，在脐带血中发现，并且 会导致暴露母亲的后代出现神经发育异常，包括早孕 体重下降、低出生体重、多动症、头围减少和行为问题。然而， PFAS暴露后的分子机制，如失调的基因和途径，特别是在 人类大脑发育的背景，仍未被探索。在这里，我们建议使用人类诱导 多能干细胞(IPSC)衍生模型，如神经前体细胞(NPC)、球体和大脑皮层 有机化合物，以填补这一知识空白。我们假设PFAS通过以下方式影响早期大脑发育 参与细胞增殖、细胞周期和生存的神经干细胞转录程序的失调。 我们还假设，这些早期的改变将对大脑皮层的形成产生重大影响 网络。我们提出了以下具体目标来检验这些假设：(1)调查剂量- PFAS对神经前体细胞周期、细胞活力和增殖的依赖性影响；(2)执行基因组- 广泛的遗传筛选与CRISPRi/a sgRNA文库，以确定修饰基因和途径 PFAS治疗；(3)在皮质器官模型中验证PFAS对长期神经发育的影响 并确定受PFAS影响的细胞群体。我们的研究将确定基因、分子和细胞途径 在体外模拟的大脑发育的不同阶段，受到PFAS暴露的失调。它有潜力 揭示PFAS暴露背后的新机制，并预测PFAS对发育中人类的影响 大脑网络功能。

SUMMARY Per- and polyfluoroalkyl substances and their derivatives (PFAS) are industrial chemicals that are wide- spread in the environment, including blood serum of wildlife and humans. Their pervasiveness and long half-life raises the questions about their toxicity, especially with regards to the effect on the developing fetus. Several studies have demonstrated that these compounds cross the placenta, are found in the umbilical cord blood, and can cause neurodevelopmental abnormalities in the offspring of the exposed mothers, including early pregnancy loss, low birth weight, hyperactivity, decreased head circumference and behavior problems. However, the molecular mechanisms following PFAS exposure, such as dysregulated genes and pathways, especially in the context of human brain development, remain unexplored. Here, we are proposing to use human induced pluripotent stem cell (iPSC) derived models such as neural progenitor cells (NPCs), spheroids and brain cortical organoids, to fill in this knowledge gap. We hypothesize that PFAS impact early brain development by dysregulating transcriptional programs of NPCs that are involved in proliferation, cell cycle and survival. We also hypothesize that these early alterations will have significant impact on the formation of cortical networks. We are proposing the following Specific Aims to test these hypotheses: (1) Investigate dose- dependent impact of PFAS on neural progenitor cell cycle, cell viability and proliferation; (2) Perform genome- wide pooled genetic screens with CRISPRi/a sgRNA libraries to identify modifier genes and pathways upon PFAS treatment; (3) Validate the impact of PFAS on long-term neurodevelopment in cortical organoids models and identify cell populations impacted by PFAS. Our study will identify genes, molecular and cellular pathways dysregulated by PFAS exposure across various stages of brain development, modeled in vitro. It has a potential to uncover new mechanisms behind PFAS exposure, and to predict the impact of PFAS on developing human brain network function.