FUS-ALS 诱导多能干细胞衍生的星形胶质细胞通过毒性获得和支持丧失机制损害人类运动单位。

FUS-ALS hiPSC-derived astrocytes impair human motor units through both gain-of-toxicity and loss-of-support mechanisms.

机构信息

Department of Neurosciences, Experimental Neurology and Leuven Brain Institute, KU Leuven - University of Leuven, 3000, Leuven, Belgium.

VIB Center for Brain & Disease Research, Laboratory of Neurobiology, 3000, Leuven, Belgium.

出版信息

Mol Neurodegener. 2023 Jan 18;18(1):5. doi: 10.1186/s13024-022-00591-3.

DOI:10.1186/s13024-022-00591-3

PMID:36653804

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9847053/

Abstract

BACKGROUND

Astrocytes play a crucial, yet not fully elucidated role in the selective motor neuron pathology in amyotrophic lateral sclerosis (ALS). Among other responsibilities, astrocytes provide important neuronal homeostatic support, however this function is highly compromised in ALS. The establishment of fully human coculture systems can be used to further study the underlying mechanisms of the dysfunctional intercellular interplay, and has the potential to provide a platform for revealing novel therapeutic entry points.

METHODS

In this study, we characterised human induced pluripotent stem cell (hiPSC)-derived astrocytes from FUS-ALS patients, and incorporated these cells into a human motor unit microfluidics model to investigate the astrocytic effect on hiPSC-derived motor neuron network and functional neuromuscular junctions (NMJs) using immunocytochemistry and live-cell recordings. FUS-ALS cocultures were systematically compared to their CRISPR-Cas9 gene-edited isogenic control systems.

RESULTS

We observed a dysregulation of astrocyte homeostasis, which resulted in a FUS-ALS-mediated increase in reactivity and secretion of inflammatory cytokines. Upon coculture with motor neurons and myotubes, we detected a cytotoxic effect on motor neuron-neurite outgrowth, NMJ formation and functionality, which was improved or fully rescued by isogenic control astrocytes. We demonstrate that ALS astrocytes have both a gain-of-toxicity and loss-of-support function involving the WNT/β-catenin pathway, ultimately contributing to the disruption of motor neuron homeostasis, intercellular networks and NMJs.

CONCLUSIONS

Our findings shine light on a complex, yet highly important role of astrocytes in ALS, and provides further insight in to their pathological mechanisms.

摘要

背景

星形胶质细胞在肌萎缩侧索硬化症（ALS）的选择性运动神经元病变中发挥着至关重要但尚未完全阐明的作用。除了其他职责外，星形胶质细胞还提供重要的神经元稳态支持，但在 ALS 中，这种功能受到严重损害。完全建立人类共培养系统可用于进一步研究功能障碍细胞间相互作用的潜在机制，并有可能为揭示新的治疗切入点提供平台。

方法

在这项研究中，我们从 FUS-ALS 患者中鉴定了人诱导多能干细胞（hiPSC）衍生的星形胶质细胞，并将这些细胞纳入人类运动单位微流控模型中，通过免疫细胞化学和活细胞记录来研究星形胶质细胞对 hiPSC 衍生的运动神经元网络和功能神经肌肉接头（NMJ）的影响。FUS-ALS 共培养物与 CRISPR-Cas9 基因编辑的同基因对照系统进行了系统比较。

结果

我们观察到星形胶质细胞稳态失调，导致 FUS-ALS 介导的炎症细胞因子反应性和分泌增加。在与运动神经元和肌管共培养时，我们检测到对运动神经元-神经突生长、NMJ 形成和功能的细胞毒性作用，同基因对照星形胶质细胞可改善或完全挽救这种作用。我们证明 ALS 星形胶质细胞具有毒性增益和支持功能丧失的双重作用，涉及 WNT/β-catenin 通路，最终导致运动神经元稳态、细胞间网络和 NMJ 的破坏。