Human IPSC-Derived PreBötC-Like Neurons and Development of an Opiate Overdose and Recovery Model.
Human IPSC-Derived PreBötC-Like Neurons and Development of an Opiate Overdose and Recovery Model.
复制标题
人类 IPSC 衍生的 PreBötC 样神经元以及阿片类药物过量和恢复模型的开发。
DOI：
10.1002/adbi.202300276
复制
发表时间：
2023
期刊：
影响因子：
3.7
通讯作者：
Hickman,JamesJ
中科院分区：
文献类型：
--
作者：
Guo,Xiufang;Akanda,Nesar;Fiorino,Gabriella;Nimbalkar,Siddharth;Long,ChristopherJ;Colón,Alisha;Patel,Aakash;Tighe,PatrickJ;Hickman,JamesJ
阿片类药物过量是药物过量致死的主要原因，迫切需要进行调查。吸气节律调节和阿片诱导的呼吸抑制（OIRD）的关键脑区是前Bötinger复合体（preBötC），目前的知识主要来自动物系统。本研究旨在建立一种从诱导多能细胞（iPSC）产生人类preBötC神经元的方案，并利用这些iPSC‐preBötC神经元开发阿片类药物过量和恢复模型。建立了从人iPSC分化preBötC样神经元的从头方案。通过免疫细胞化学分析，这些神经元表达必需的preBötC标志物，并通过膜片钳电生理学分析，证明了对preBötC调节剂的预期电生理反应。特异性生物标志物和功能分析的相关性强烈提示了前BötC样表型。此外，证明了四种不同阿片类药物对这些神经元活性的剂量依赖性抑制，其确定的IC50与文献相当。纳洛酮以浓度依赖性方式挽救抑制。这种iPSC‐preBötC模拟物对于研究OIRD和对抗过量危机至关重要，也是将功能性过量模型整合到微生理系统中的第一步。
Opioid overdose is the leading cause of drug overdose lethality, posing an urgent need for investigation. The key brain region for inspiratory rhythm regulation and opioid‐induced respiratory depression (OIRD) is the preBötzinger Complex (preBötC) and current knowledge has mainly been obtained from animal systems. This study aims to establish a protocol to generate human preBötC neurons from induced pluripotent cells (iPSCs) and develop an opioid overdose and recovery model utilizing these iPSC‐preBötC neurons. A de novo protocol to differentiate preBötC‐like neurons from human iPSCs is established. These neurons express essential preBötC markers analyzed by immunocytochemistry and demonstrate expected electrophysiological responses to preBötC modulators analyzed by patch clamp electrophysiology. The correlation of the specific biomarkers and function analysis strongly suggests a preBötC‐like phenotype. Moreover, the dose‐dependent inhibition of these neurons’ activity is demonstrated for four different opioids with identified IC50's comparable to the literature. Inhibition is rescued by naloxone in a concentration‐dependent manner. This iPSC‐preBötC mimic is crucial for investigating OIRD and combating the overdose crisis and a first step for the integration of a functional overdose model into microphysiological systems.