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

摘要 此补充申请是为了在过渡到之前实现之前修改的里程碑 UH3阶段。这项补充资金请求是由于进展受到严重阻碍而产生的结果 在新冠肺炎大流行期间，业务能力受到限制。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招致额外的人员 以及供应成本。 该项目的总体意义和范围与原提案相同。有一个 迫切需要开发治疗阿片类药物使用障碍(OUD)的疗法，同时也需要产生疼痛 不会上瘾的治疗方法。这两个目标都将通过符合以下条件的高吞吐量模型实现 基于人类细胞的药物筛选，概括了成瘾者潜在的神经生物学特征 进程。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 和中脑的GABA能神经元，长期以来被认为负责调节增强 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)来源的中脑的微生理系统 3维平台上的多巴胺(DA)/伽马氨基丁酸(GABA)神经元，最终将 结合小胶质细胞、血脑屏障和肝脏代谢成分。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 慢性阿片类药物诱导的DA反应性可塑性被认为是1)快感缺失的基础 阿片类药物戒断的特点；2)慢性疼痛状态的负面情感成分；3)渴望和 旧病复发。模型的慢性疼痛相关方面将通过检查厌恶的kappa- 除常用的Mu阿片受体激动剂外，阿片类药物对DA传递的介导作用， 并通过在模型中加入炎症介导性小胶质细胞。吞吐量将增加 将在线传感器集成到MPS中，用于在线检测DA。

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.