Department of Bioengineering, University of Washington, Seattle, WA, 98195, USA.
Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA, 98109, USA; Department of Physiology & Biophysics, University of Washington, Seattle, WA, 98195, USA.
Biomaterials. 2021 Apr;271:120700. doi: 10.1016/j.biomaterials.2021.120700. Epub 2021 Feb 3.
Neurons derived from human induced pluripotent stem cells (hiPSCs) are powerful tools for modeling neural pathophysiology and preclinical efficacy/toxicity screening of novel therapeutic compounds. However, human neurons cultured in vitro typically do not fully recapitulate the physiology of the human nervous system, especially in terms of exhibiting morphological maturation, longevity, and electrochemical signaling ability comparable to that of adult human neurons. In this study, we investigated the potential for astrocyte-derived extracellular vesicles (EVs) to modulate survival and electrophysiological function of human neurons in vitro. Specifically, we demonstrate that EVs obtained from human astrocytes promote enhanced single cell electrophysiological function and anti-apoptotic behavior in a homogeneous population of human iPSC-derived cortical neurons. Furthermore, EV-proteomic analysis was performed to identify cargo proteins with the potential to promote the physiological enhancement observed. EV cargos were found to include neuroprotective proteins such as heat shock proteins, alpha-synuclein, and lipoprotein receptor-related protein 1 (LRP1), as well as apolipoprotein E (APOE), which negatively regulates neuronal apoptosis, and a peroxidasin homolog that supports neuronal oxidative stress management. Proteins that positively regulate neuronal excitability and synaptic development were also detected, such as potassium channel tetramerization domain containing 12 (KCTD12), glucose-6- phosphate dehydrogenase (G6PD), kinesin family member 5B (KIF5B), spectrin-alpha non-erythrocytic1 (SPTAN1). The remarkable improvements in electrophysiological function and evident inhibition of apoptotic signaling in cultured neurons exposed to these cargos may hold significance for improving preclinical in vitro screening modalities. In addition, our collected data highlight the potential for EV-based therapeutics as a potential class of future clinical treatment for tackling inveterate central and peripheral neuropathies.
神经元衍生自人类诱导多能干细胞（hiPSCs），是模拟神经病理生理学和新型治疗化合物的临床前疗效/毒性筛选的有力工具。然而，在体外培养的人类神经元通常不能完全再现人类神经系统的生理学，尤其是在形态成熟、寿命和与成年人类神经元相当的电化学信号传递能力方面。在这项研究中，我们研究了星形胶质细胞衍生的细胞外囊泡（EVs）在体外调节人类神经元的存活和电生理功能的潜力。具体而言，我们证明来自人类星形胶质细胞的 EV 促进了同质群体的人类 iPSC 衍生皮质神经元的单细胞电生理功能增强和抗凋亡行为。此外，还进行了 EV 蛋白质组学分析，以鉴定具有促进观察到的生理增强潜力的载体蛋白。EV 货物包括神经保护蛋白，如热休克蛋白、α-突触核蛋白和脂蛋白受体相关蛋白 1（LRP1），以及负调节神经元凋亡的载脂蛋白 E（APOE）和支持神经元氧化应激管理的过氧化物酶同源物。还检测到了正向调节神经元兴奋性和突触发育的蛋白质，如钾通道四聚化结构域包含 12（KCTD12）、葡萄糖-6-磷酸脱氢酶（G6PD）、驱动蛋白家族成员 5B（KIF5B）、非红细胞 spectrin-alpha 1（SPTAN1）。暴露于这些货物的培养神经元的电生理功能显著改善，凋亡信号明显抑制，这可能对改善临床前体外筛选方法具有重要意义。此外，我们收集的数据突出了基于 EV 的治疗方法的潜力，作为一种潜在的未来临床治疗方法，用于解决顽固的中枢和周围神经病变。
