Spatio-temporal dissection of epithelial cell hierarchies in gut inflammation

肠道炎症中上皮细胞层次结构的时空解剖

• 批准号: 9479946
• 负责人: Ken S Lau
• 金额: $ 2.63万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-07 至 2020-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9479946
• 关键词: Acute; Affect; Algorithms; Alpha Cell; Architecture; Behavior; Biology; Cell Differentiation process; Cell Lineage; Cells; Chronic; Combined Modality Therapy; Complex; Computer Analysis; Crohn' s disease of the ileum; Cytometry; Data; Development; Diagnosis; Differentiation Antigens; Disease; Dissection; Distal part of ileum; Emerging Technologies; Epithelial; Epithelial Cells; Epithelium; Etiology; Foundations; Future; GTP-Binding Protein alpha Subunits, Gs; Homeostasis; Human; Image; Immune; Immune system; Individual; Inflammation; Inflammatory; Inflammatory Bowel Diseases; Inflammatory disease of the intestine; Intestinal Mucosa; Intestines; Knock-out; Maintenance; Microscopy; Modeling; Modification; Molecular; Multipotent Stem Cells; Mus; Outcome; Outcomes Research; Paneth Cells; Pathologic; Pathology; Pathway interactions; Patients; Plasticizers; Process; Recruitment Activity; Research; Resolution; Risk Factors; Role; Sampling; Signal Pathway; Signal Transduction; Signaling Protein; Stem cells; Structure; System; Systems Biology; T-Lymphocyte; T-Lymphocyte Subsets; TNF gene; Technology; Time; Tissues; Villus; Work; base; cell type; cytokine; direct application; gamma secretase; ileum; in vivo; inhibitor/antagonist; insight; intestinal epithelium; intraepithelial; new therapeutic target; notch protein; novel; organizational structure; patient stratification; predicting response; predictive modeling; programs; public health relevance; response; single cell analysis; spatiotemporal

 DESCRIPTION (provided by applicant): We use state-of-the-art technologies and computational analyses to understand, at single cell resolution, how the organization of epithelial differentiation is altered by selective perturbations or inflammatory perturbations of te terminal ileum. During the continuous homeostatic renewal process, an epithelial progenitor cell progressively differentiates according to a set of rules set forth by a composite of signaling pathways that are spatially arranged along the crypt-villus axis. Decades of intestinal biology research have established that differentiation proceeds stepwise, with branched decisions resulting in a hierarchical organization of the different functional lineages. How individual cells coherently integrate multiple signaling pathways into a hierarchical differentiation program, and how this program is coordinately altered when perturbed in states of disease are undefined. In inflammatory bowel disease (IBD), the organized intestinal microenvironment is infiltrated by a variety of immune cells that provide additional signaling inputs to epithelial cells. We hypothesize that normal signaling organization is perturbed by these additional inputs, which in turn, either rearranges or abolishes the hierarchical organization of epithelial differentiation, leading to imprecise differentiation, failed maintenance of the mature state, and altered proportions of epithelial cell types. We will coordinate two emerging technologies, MultiOmyx microscopy and Cytometry Time-of-Flight (CyTOF), to interrogate multiple signaling activities and cell differentiation pathways at single cell resolution, in order to use data-driven computational approaches to model differentiation hierarchies. In the two aims of this proposal, we will apply our approaches to study (Aim 1) the effects of specific pathway inhibition, which we hypothesize to maintain hierarchical differentiation, and (Aim 2) the effects of chronic inflammation, which we hypothesize to convert the strict differentiation hierarchy into a plastic landscape. The study of stromal and immune cell contributions to the organization of differentiation will be studied using a combination of enteroid cultures, live imaging, and multiplex microscopy of tissue sections. Future directions include the direct application of our experimental-computational platforms to human patient samples of Crohn's ileitis. Through our findings, we aim to make significant contributions to the understanding, diagnosis and treatment of IBD. The outcomes from this research will hopefully be a gateway towards the study of other complex diseases with undefined etiologies, molecular distribution of many risk factors over many components, and interactions between multiple heterologous systems.

 描述（由适用提供）：我们使用最先进的技术和计算分析来理解，在单细胞分辨率下，如何通过选择性的扰动或TE末端回肠的炎症性扰动来改变上皮分化的组织。在连续的稳态更新过程中，上皮祖细胞根据一组由沿加密villus轴空间布置的信号通路列出的规则逐渐区分。数十年的肠生物学研究已经确定，分化逐步进行，分支决策导致了不同功能谱系的分层组织。单个细胞如何 连贯地整合了多个信号通路纳入分层分化程序，以及在疾病状态中扰动时该程序如何协调一致。在炎症性肠病（IBD）中，有组织的肠道微环境被多种免疫电池浸润，这些免疫力为上皮细胞提供了其他信号输入。我们假设正常信号组织受到这些其他输入的干扰，而这些输入反过来又重新排列或废除了上皮分化的分层组织，从而导致差异分化，对成熟状态的维持失败，并改变了上皮细胞类型的比例。我们将协调两种新兴技术，即多玛莎显微镜和流动时间（CYTOF），以询问单细胞分辨率下的多个信号传导活性和细胞分化途径，以便使用数据驱动的计算方法来模拟分化层次结构。在该提案的两个目的中，我们将应用我们的方法研究（目标1）特定途径抑制的影响，我们假设这是维持层次分化的，以及（AIM 2）慢性感染的影响，我们假设这些影响将严格的分化等级转化为塑料景观。将研究对分化组织的基质和免疫细胞对分化组织的贡献的研究。未来的方向包括将我们的实验计算平台直接应用于克罗恩病炎的人类患者样本。通过我们的发现，我们旨在为IBD的理解，诊断和治疗做出重大贡献。这项研究的结果有望成为研究其他具有不确定病因的复杂疾病，许多危险因素在许多组件上的分子分布以及多个异源系统之间的相互作用的门户。