Neonatal gut-on-a-chip platform for high content drug testing and precision medicine

项目总结/摘要 该提案的目标是研究人类微生理学芯片平台作为一种生物学方法的作用。 一种精确的医学工具，可以模拟一种影响早产儿的毁灭性疾病， 小肠结肠炎（NEC）。我们使用类器官和“芯片上NEC”模型开发了NEC的临床前模型 系统在体外重现人类疾病的肠道环境，获得对疾病的新见解 发病机制，并测试我们的模型的功能和临床效用，以评估候选物的功效。 治疗学我们的NEC芯片模型利用早产儿肠道类器官沿着 人内皮细胞和患者来源的微生物群，以重建早产肠道的关键方面， 病理生理学我们的初步研究表明，这些成分在caseine-on-a- 芯片微流控装置产生在人类NEC中观察到的临床特征，例如肠道屏障衰竭， 细胞紧密连接的破坏，上皮细胞增殖的减少，前- 炎性细胞因子应答以及显著量的细胞死亡。在本建议中，我们将使用几个 多组学方法来表征我们的NEC-on-a-chip模型并与人NEC表型进行比较。 为了实现这一目标，我们开发了一个多中心NEC生物储存库，其中包括详细的临床元数据 对应于过多的人类样本，包括从大肠杆菌的活组织检查培养的肠类器官。 有或无NEC的早产儿。此外，我们还创造了一种高通量和高含量的药物， 筛选平台，使用未成熟的肠类器官来鉴定抑制肠类器官的药物或化合物。 体外观察到的致病性炎症反应。此外，我们将证明功能和临床实用性 我们的患者衍生的NEC芯片模型作为一个精确的医学平台，以测试剂量，疗效， 候选治疗剂的毒性。为了成功完成这些研究，我们建立了一个多学科的 团队由新生儿学家、细胞生物学家、儿科外科医生、基因组科学家和 生物信息学家。综合起来，这些研究将在我们的理解中取得重大的概念性进展 与发育中的早产儿肠道微生物组的多细胞相互作用，并提供新的模型 系统和临床前平台，通过这些系统和平台， 肠道疾病可以在这个脆弱的患者群体中进行。

Project Summary/Abstract The goal of this proposal is to investigate the role of a human microphysiologic intestine-on-a-chip platform as a precision medicine tool to model a devastating disease affecting premature infants known as necrotizing enterocolitis (NEC). We developed preclinical models of NEC using both organoids and a “NEC-on-a-chip” model system to recapitulate the intestinal environment of the human disease in vitro, gain new insights into disease pathogenesis and test the functional and clinical utility of our models to evaluate the efficacy of candidate therapeutics. Our NEC-on-a-chip model utilizes a combination of premature infant intestinal organoids along with human endothelial cells and patient-derived microbiota, to recreate critical aspects of premature gut pathophysiology. Our preliminary studies demonstrate that co-culture of these components on intestine-on-a- chip microfluidic devices produces clinical features seen in human NEC such as gut barrier failure with the breakdown of cellular tight junctions, decreased epithelial cell proliferation, a dramatic increase in the pro- inflammatory cytokine response, as well as a significant amount of cell death. In this proposal, we will use several multi-omic approaches to characterize our NEC-on-a-chip model and compare to the human NEC phenotype. To achieve this, we developed a multi-center NEC Biorepository, which consists of detailed clinical metadata corresponding to a plethora of human specimens, including intestinal organoids cultured from the biopsies of premature infants with or without NEC. Furthermore, we have created a high-throughput and high-content drug screening platform using premature intestinal organoids to identify drugs or compounds that inhibit the pathogenic inflammatory responses seen in vitro. Moreover, we will demonstrate the functional and clinical utility of our patient-derived NEC-on-a-chip model as a precision medicine platform to test the dosing, efficacy, and toxicity of candidate therapeutics. To successfully complete these studies, we established a multi-disciplinary team with the expertise of a Neonatologist, Cell Biologist, Pediatric Surgeon, Genome Scientist and Bioinformatician. Taken together, these studies will make a significant conceptual advance in our understanding of the multicellular interactions with the microbiome of the developing premature intestine and provide new model systems and preclinical platforms by which the identification and testing of therapeutics for NEC and other intestinal diseases can be performed in this vulnerable patient population.