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

摘要 阿片类药物滥用所带来的公共卫生危机的意义是非常明显的。 迫切需要开发治疗阿片类药物使用障碍(OUD)的疗法，也需要 开发不会上瘾的疼痛治疗方法。这两个目标都将通过高吞吐量模型实现 基于人类细胞的药物筛选，概括了神经生物学的特征 在令人上瘾的过程中。我们建议开发的模型集中在成瘾的一个关键成分上 回路--中脑的多巴胺能和γ-氨基丁酸(GABA)能神经元 被认为对调节许多类滥用药物的强化特性负有责任，包括 阿片类药物。我们将开发基于人类诱导的多器官、微生理系统(MPSS) 多能干细胞(IPSC)来源的中脑命运多巴胺(DA)/GABA神经元的三维结构 包含小胶质细胞、血脑屏障(BBB)和肝脏代谢组件的平台。核糖核酸 测序(RNAseq)和代谢组学分析将补充主要的DA释放措施，以识别 慢性阿片类药物诱导DA反应性可塑性的新机制被认为是基础1) 阿片类药物戒断的快感缺失特征：2)慢性疼痛状态的负性情感成分； 3)渴求和故态复萌。模型的慢性疼痛相关方面将通过检查 除通常滥用的MU阿片外，厌恶kappa介导的阿片类药物对DA传递的影响 受体激动剂，并通过在模型中加入炎症中介小胶质细胞。成立为法团 血脑屏障和肝脏代谢模块进入MPS平台将允许筛选UH3阶段的药物 说明滥用阿片类药物不同给药途径和潜在生物利用度的项目 治疗药物，增加可译性。通过集成在线传感器，将提高吞吐量 用于项目第三阶段确定的DA和其他关键分析物的在线检测。为 我们将使用加州大学洛杉矶分校精心挑选的100个化学基因组探针进行成瘾治疗筛查 激酶抑制物库。对激酶的关注是基于它们在可塑性中所扮演的角色，并将涵盖 沿着BDNF信号通路的活动，mTORC2、mTORC1、AKT等靶点沿着BDNF轴。 我们在这方面对化合物/靶点的选择也将受到RNAseq和代谢组学的影响 UG3阶段的结果，如果加州大学洛杉矶分校的分子筛查需要，我们将补充其他药物 共享资源化学基因组文库，其中有3000多个探针强大。虽然没有在这里提出，但它 还需要指出的是，MOC设备也可以用来测试小说的滥用责任 由其他筛查工具确定的抗伤害感受剂。拥有一个跨学科的科学家团队和 工程师们，这项提议将建立在芯片器官和IPSC技术的最新进展基础上，以创造 创新的MPSS具有很高的潜力，在未来将产生重大临床影响。

Abstract The significance of the public health crisis presented by the epidemic in opioid abuse is abundantly clear. 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 the use of human cells, that recapitulate features of the neurobiology underlying the addictive process. The model we propose to develop focuses on a key component of addictive circuitry – the dopaminergic and Gamma-Amino Butyric Acid (GABA)ergic neurons of the midbrain, long recognized as responsible for mediating the reinforcing properties of many classes of abused drugs, including opioids. We will develop multi-organ, microphysiological systems (MPSs) based on the use of human induced pluripotent stem cell (iPSC)-derived midbrain-fated dopamine (DA)/ GABA neurons on a three-dimensional platform that incorporates microglia, blood-brain-barrier (BBB) 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. The incorporation of BBB and liver metabolism modules into the MPS platform will permit screening of drugs in the UH3 phase of the project that accounts for different routes of abused opioid administration and the bioavailability of potential therapeutic drugs, increasing translatability. Throughput will be increased by the integration of online sensors into the MPS for online detection of DA and other key analytes identified in the UG3 phase of the project. For addiction-treatment screening we will use a curated set of 100 chemical genomics probes from our UCLA kinase inhibitor library. The focus on kinases is based on their well-described role in plasticity, and will cover activities along the BDNF signaling pathway, mTORC2, mTORC1, AKT and other targets along the BDNF axis. Our selection of compounds/targets in this context will also be informed by the RNAseq and metabolomics results from the UG3 phase and we will supplement other drugs if needed from UCLA’s Molecular Screening Shared Resource chemical genomics libraries, which is over 3k probes strong. Although not proposed here, it is also important to point out that the MoC device could also be used to test the abuse liability of novel antinociceptive agents identified by other screening tools. With an interdisciplinary team of scientists and engineers, this proposal will build upon recent advances in organ-on-a-chip and iPSC technologies to create innovative MPSs with high potential to have significant clinical impacts in the future.