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

项目摘要/摘要 这项提议的目标是调查人类微生理芯片上的肠平台作为 一种精确的医学工具，用于模拟一种称为坏死性早产儿的破坏性疾病 小肠炎(NEC)。我们使用有机化合物和“芯片上的NEC”模型开发了NEC的临床前模型。 系统在体外概括了人类疾病的肠道环境，对疾病有了新的见解 并测试我们的模型的功能和临床实用性，以评估候选的疗效 治疗学。我们的NEC芯片上模型利用了早产儿肠道器官和 人内皮细胞和患者来源的微生物区系，以重建早产儿肠道的关键方面 病理生理学。我们的初步研究表明，这些成分在肠-on-a-a上的共培养 芯片微流控设备可产生人类NEC的临床特征，如肠道屏障功能障碍 细胞紧密连接破裂，上皮细胞增殖减少，细胞内亲和力显著增强。 炎性细胞因子反应，以及大量细胞死亡。在这个提案中，我们将使用几个 多组学方法来描述我们的芯片上NEC模型，并与人类NEC表型进行比较。 为了实现这一点，我们开发了一个多中心NEC生物信息库，它包含详细的临床元数据 与过多的人类样本相对应，包括从猪的活检组织中培养的肠道器官 患有或不患有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 使用未成熟的肠道有机化合物进行筛选的平台，以识别抑制该过程的药物或化合物 在体外可见致病性炎症反应。此外，我们还将展示其功能和临床应用。 将我们患者衍生的NEC芯片模型作为精密医学平台，以测试剂量、疗效和 候选治疗药物的毒性。为了顺利完成这些研究，我们建立了一个多学科的 团队拥有新生儿专家、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。综上所述，这些研究将在我们的理解上取得重大的概念性进展 多细胞与发育中的早产儿肠道微生物群的相互作用并提供新的模型 用于识别和测试NEC和其他药物的系统和临床前平台 肠道疾病可以在这一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.