Multi-organ-on-chip device for modeling opioid reinforcement and withdrawal, and the negative affective component of pain: a therapeutic screening tool.

总结 此补充申请旨在帮助在过渡到之前完成先前修改的里程碑 UH 3阶段。之所以要求提供补充资金，是因为进展遇到重大障碍， 于COVID-19疫情期间营运能力受到限制。虽然取得了重大进展， 完成里程碑所需的额外时间将导致我们需要额外的人员 和供应成本。 该项目的总体意义和范围与原提案相同。有一个 迫切需要开发用于治疗阿片类药物使用障碍（OUD）的治疗剂，并且还需要开发用于治疗疼痛的药物。 非成瘾性的治疗。这两个目标都将通过高通量模型来实现， 基于人类细胞的药物筛选，概括了成瘾性疾病背后的神经生物学特征， 过程我们正在开发的模型专注于成瘾回路的一个关键组成部分-多巴胺能神经元。 和中脑的GABA能神经元，长期以来被认为是负责介导增强 包括阿片类药物在内的许多类滥用药物的性质。我们正在开发一种多器官 基于人诱导多能干细胞（iPSC）衍生的中脑的微生理系统（MPS） 多巴胺（DA）/γ-氨基丁酸（GABA）神经元在三维平台上，最终将 整合了小胶质细胞、血脑屏障和肝代谢成分。RNA测序（RNAseq）和 代谢组学分析将补充主要的DA释放措施，以确定新的机制 有助于慢性阿片诱导的DA反应性可塑性，被认为是1）快感缺乏的基础 阿片类药物戒断的特征; 2）慢性疼痛状态的负面情感成分; 3）渴望和 复发模型的慢性疼痛相关方面将通过检查厌恶性kappa- 除了通常滥用的μ阿片受体激动剂外， 以及通过将炎症介导的小胶质细胞掺入模型中。将通过增加 将在线传感器集成到MPS中，用于DA的在线检测。

SUMMARY This supplemental application is to facilitate fulfillment of previously modified milestones prior to transition to the UH3 phase. This request for supplemental funds is a result of significant impediments to progress resulting from restriction of operational capacity during the COVID-19 pandemic. While significant progress has been made, the additional time required to complete the milestones will result in us incurring additional personnel and supply costs. The overall significance and scope of the project remains the same as the original proposal. There is a desperate need to develop therapeutics for treatment of opioid use disorder (OUD), and also to develop pain treatments that are non-addictive. Both of these goals will be served by high-throughput models amenable to drug screening, based on human cells, that recapitulate features of the neurobiology underlying the addictive process. The model we are developing focuses on a key component of addictive circuitry – the dopaminergic and GABAergic neurons of the midbrain, long recognized as responsible for mediating the reinforcing properties of many classes of abused drugs, including opioids. We are developing a multi-organ microphysiological system (MPS) based on human induced pluripotent stem cell (iPSC)-derived midbrain dopamine (DA)/Gamma-Amino Butyric Acid (GABA) neurons on a 3-dimensional platform that ultimately will incorporate microglia, blood-brain-barrier and liver metabolism components. RNA sequencing (RNAseq) and metabolomics analyses will complement the primary DA release measure to identify novel mechanisms contributing to chronic opioid-induced plasticity in DA responsiveness thought to underlie 1) the anhedonia characteristic of opioid withdrawal; 2) the negative affective component of chronic pain states; 3) craving and relapse. The chronic pain-relevant aspect of the model will be realized by examination of aversive kappa- mediated opioid effects on DA transmission in addition to the commonly abused mu opioid receptor agonists, and by incorporation of inflammatory-mediating microglia into the model. Throughput will be increased by the integration of online sensors into the MPS for online detection of DA.