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

研究全氟烷基酸及其衍生物在人脑类器官模型中的神经发育毒性

• 批准号: 10563204
• 负责人: LILIA M IAKOUCHEVA
• 金额: $ 60.57万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-07 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10563204
• 关键词: 3-Dimensional; ATAC-seq; Acids; Affect; Animals; Astrocytes; Attention deficit hyperactivity disorder; Biological; Blood - brain barrier anatomy; Blood specimen; Brain; CRISPR interference; Carbon; Carboxylic Acids; Carpet; Cell Cycle; Cell Differentiation Induction; Cell Proliferation; Cell Survival; Cells; Chemicals; Chromatin; Clothing; Complex; Detection; Development; Developmental Delay Disorders; Disease; Dose; Environment; Environmental Risk Factor; Experimental Models; Family; Fetal Development; Fluorine; Food; Genes; Genetic; Genetic Screening; Genetic Transcription; Glutamates; Growth; Half-Life; Head circumference; High Prevalence; Human; Human Development; Hyperactivity; Industrialization; Knowledge; Libraries; Literature; Low Birth Weight Infant; Machine Learning; Microcephaly; Modeling; Molecular; Morphology; Mothers; Mus; Neurodevelopmental Disorder; Neuroglia; Neurons; Organoids; Paper; Pathway interactions; Phenotype; Placenta; Play; Poly-fluoroalkyl substances; Population; Prevalence; Problem behavior; Proliferating; Rattus; Reporting; Role; Schizophrenia; Serum; Sulfonic Acids; Testing; Time; Toxic effect; Umbilical Cord Blood; Zebrafish; autism spectrum disorder; behavioral phenotyping; developmental toxicity; differential expression; early pregnancy loss; excitatory neuron; exome; exposed human population; gene discovery; genetic risk factor; genome sequencing; genome wide association study; genome-wide; human stem cells; in vitro Model; induced pluripotent stem cell; insight; lentivirally transduced; man; multi-electrode arrays; nerve stem cell; neural network; neurodevelopment; offspring; postnatal; progenitor; programs; rare variant; single-cell RNA sequencing; stem cell model; transcriptome sequencing; transcriptomics; variant detection; whole genome

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.

总结 全氟烷基和多氟烷基物质及其衍生物（PFAS）是工业化学品， 在环境中传播，包括野生动物和人类的血清。它们的普遍性和长半衰期 提出了关于其毒性的问题，特别是关于对发育中的胎儿的影响。几 研究表明，这些化合物可以穿过胎盘，存在于脐带血中， 可导致暴露于此的母亲的后代神经发育异常，包括早孕 体重减轻、出生体重低、多动、头围减小和行为问题。但 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下，仍然未被探索。在这里，我们建议使用人类诱导 多能干细胞（iPSC）衍生的模型，例如神经祖细胞（NPC）、球状体和脑皮质细胞（BMSCs）。 类器官，来填补这一知识空白。我们假设PFAS通过以下方式影响早期大脑发育： 参与增殖、细胞周期和存活的NPC的转录程序失调。 我们还假设，这些早期改变将对皮质骨的形成产生重大影响。 网络.我们提出以下具体目标来检验这些假设：（1）研究剂量- PFAS对神经前体细胞周期、细胞活力和增殖的依赖性影响;（2）进行基因组- 使用CRISPRi/a sgRNA文库进行广泛的合并遗传筛选，以鉴定修饰基因和途径， PFAS治疗;（3）观察PFAS对皮质类器官模型长期神经发育的影响 并识别受PFAS影响的细胞群。我们的研究将识别基因、分子和细胞途径 通过PFAS暴露在大脑发育的各个阶段失调，在体外建模。它有潜力 揭示PFAS暴露背后的新机制，并预测PFAS对发育中的人类的影响。 脑网络功能