hiPSC-based DRG Tissue Mimics on Multi-well Microelectrode Arrays as a Tissue Chip Model of Acute and Chronic Nociception

项目总结/摘要： 慢性疼痛折磨着多达五分之一的成年人，并且是美国长期残疾的最常见原因。 世界阿片类药物通常用于非癌症疼痛，与高发病率相关。 严重影响和滥用。此外，目前用于研究伤害感受和测试的体内和体外模型 潜在的治疗是不够的。基于人类的，病理学相关的伤害感受模型迫切需要 需要促进新的非阿片类疼痛疗法的临床前开发。因此，我们建议 使用hiPSC感觉神经元和卫星开发急性和慢性伤害感受的创新3D模型 神经胶质细胞替代物（基于hiPSC的DRG组织模拟物）在多孔MEA上的生长。 在UG 3阶段，我们将开发一种基于3D的组织芯片，用于模拟急性和慢性伤害感受。 基于hiPSC背根神经节（DRG）组织模拟物和高含量、中等通量微电极 阵列（MEA）平台。DRG组织模拟物将由hiPSC对应物组成，以组成神经节内神经节。 DRG细胞类型嵌入胶原基质中。然后我们将展示稳定的自发性和有害性 刺激诱发的行为响应于热、化学和电刺激挑战。此外，委员会认为， 我们的目标是证明利用3D混合细胞DRG组织的明显功能和表型优势 模拟与纯神经元2D或3D模型。更具体地说，我们的目标是证明敏感性， 通过神经元和非神经元细胞类型之间的配体受体相互作用进行翻译控制，从而 证明与疼痛的“神圣Trinity”（伤害性、炎症性和神经性）的病理相关性 以及我们的模型测试与非神经元支持的贡献相关的基本假设的能力 细胞在慢性疼痛的发展和维持。 在UH 3阶段，我们将展示我们的产品强大的定量效率和临床前疗效。 通过检测已知的基于配体的翻译控制和电压调节剂， 门控离子通道拮抗剂在慢性伤害感受的致敏模型中的作用。这两类药物是 被广泛认为是用于逆转伤害感受可塑性和/或用作外周免疫调节剂的候选化合物。 止痛药此外，我们将根据广泛接受的测定评分定量定义药理学命中 方法论。最后，我们将利用高通量性质或我们的组织芯片模型来筛选FDA- 获得批准的生物活性化合物，证明了我们高含量检测的灵敏度和通量， 潜在地鉴定被不太复杂的表型分析方法掩盖的候选治疗剂的功效 筛选

Project summary/abstract: Chronic pain afflicts up to one in five adults and is the most common cause of long-term disability in the world. Opioids, which are commonly prescribed for non-cancer pain, are associated with a high incidence of serious effects and abuse. Moreover, current in vivo and in vitro models used to study nociception and test potential treatments are inadequate. Human-based, pathology-relevant models of nociception are urgently needed to facilitate preclinical development of new non-opioid pain therapeutics. Therefore, we propose to develop an innovative 3D model of acute and chronic nociception using hiPSC sensory neurons and satellite glial cell surrogates (an hiPSC-based DRG tissue mimic) on multi-well MEAs. In the UG3 phase, we will develop a tissue chip for modeling acute and chronic nociception based on 3D hiPSC-based dorsal root ganglion (DRG) tissue mimics and a high-content, moderate-throughput microelectrode array (MEA) platform. DRG tissue mimics will be comprised of hiPSC counterparts to constituent intraganglionic DRG cell types embedded in a collagen matrix. We will then demonstrate stable spontaneous and noxious stimulus-evoked behavior in response to thermal, chemical, and electrical stimulation challenges. Furthermore, we aim to demonstrate the clear functional and phenotypic advantages of utilizing a 3D mixed-cell DRG tissue mimic versus purely neuronal 2D or 3D models. More specifically, we aim to demonstrate sensitivity to translational control via ligand receptor interactions between neuronal and non-neuronal cell types, thereby demonstrating pathological relevance to a the ‘holy trinity’ of pain (nociceptive, inflammatory and neuropathic) and our model’s capacity for testing fundamental hypotheses related to contributions of non-neuronal support cells in chronic pain development and maintenance. In the UH3 phase, we will demonstrate the powerful quantitative efficiency and preclinical efficacy of our microphysiological system by detecting known ligand-based modulators of translational control and voltage- gated ion channel antagonists in a sensitized model of chronic nociception. These two classes of drugs are widely recognized as candidate compounds for reversing nociceptive plasticity and/or serving as peripheral analgesics. Moreover, we will quantitatively define pharmacological hits based on widely accepted assay scoring methodologies. Lastly, we will leverage the high-throughput nature or our tissue chip model to screen FDA- approved, bioactive compounds, demonstrating the sensitivity and throughput of our high content assay, and potentially identifying efficacy of candidate therapeutics obscured by less sophisticated methods of phenotypic screening.