Deconstructing the fibrotic microenvironment in Crohn's disease to promote tissue healing

解构克罗恩病的纤维化微环境，促进组织愈合

基本信息

批准号：
MR/X008789/1
负责人：
Eileen Gentleman
金额：
$ 78.45万
依托单位：
King's College London
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2023
资助国家：
英国
起止时间：
2023 至无数据
项目状态：
未结题

来源：
https://gtr.ukri.org/projects?ref=MR%2FX008789%2F1
关键词：
Deconstructing fibrotic microenvironment Crohn disease

项目摘要

Research context: Over half of Crohn's disease (CD) patients develop scarring (fibrosis) around the intestine which requires life-altering surgery. However, there are currently no treatments for CD that target intestinal fibrosis. We have previously shown that when we combine human intestinal organoids (mini-intestines in-a-dish, HIO) with a jelly-like material (hydrogel) that mimics the tissue surrounding the gut where fibrosis takes place (matrix), we can study gut fibrosis in the lab. Here, we hypothesise that by modulating our hydrogels to mimic the stiff, diseased CD matrix, we can use HIO to better understand how fibrosis contributes to CD in the intestinal epithelium, and identify new ways to resolve or prevent fibrosis in patients. Gold-standard anti-inflammatory treatments for CD help 2/3 of patients, but they do not prevent or reverse fibrosis. Our goal is to identify new ways to treat CD by explicitly focusing on the fibrotic matrix. Aims and objectives: We aim to use HIO and synthetic hydrogels to understand how changes in the stiffness and composition of the fibrotic matrix that surrounds the gut in CD contribute to disease. By untangling these interactions, we hope to broaden therapeutic strategies for treating CD by identifying ways to target the matrix to reverse or prevent fibrosis. To accomplish this, we will:Objective 1: Determine if physical cues play a role in driving CD-like epithelial phenotypes. We will measure the stiffness of normal and fibrotic human intestinal tissue using atomic force microscopy, and encapsulate HIO within hydrogels that mimic these and other physical cues. We will then analyse encapsulated HIO to understand how physical cues like stiffness impact signaling pathways within the HIO epithelium, and whether hydrogel stiffness and degradability alone can prompt HIO to form fibrotic-like matrix around themselves.Objective 2: Determine if matrisome cues play a role driving CD-like epithelial phenotypesWe will use mass spectrometry to profile the composition (matrisome) of normal and fibrotic human intestinal tissue to identify proteins that are more abundant in diseased tissue. We will encapsulate HIO within hydrogels that use bioengineering strategies to incorporate or sequester fibrotic matrix components, and then analyse HIO to determine if specific proteins in the diseased matrix impact HIO or if the composition of the diseased matrix prompts HIO to stiffen their local surroundings. Objective 3: Determine whether the dysregulated matrix impacts epithelial healingWe will induce damage in HIO and use stiffness- and matrix-mimicking hydrogels from Objectives 1&2 to ask if CD-like stiffness or matrix composition impact intestinal healing, and whether this is mediated by specific signalling pathways or proteins secreted by HIO. We will also use our models to test existing drugs that target the matrix to determine if they can promote intestinal healing.Potential applications and benefits: Gold-standard treatments that alleviate inflammation in CD patients only benefit 2/3 of patients and cannot reverse or prevent intestinal fibrosis. Despite this, most research in CD focuses on moderating inflammation to promote healing. Our research approach aims to take a tissue-level perspective on CD and intestinal healing by focussing on reciprocal interactions between the epithelium and the matrix. In this project, we aim to discover new ways to treat CD by focusing on the fibrotic matrix.

研究背景：超过一半的克罗恩病 (CD) 患者肠道周围会出现疤痕（纤维化），需要进行改变生活的手术。然而，目前尚无针对肠道纤维化的克罗恩病治疗方法。我们之前已经证明，当我们将人类肠道类器官（培养皿中的迷你肠，HIO）与模拟发生纤维化的肠道周围组织（基质）的果冻状材料（水凝胶）结合起来时，我们可以研究实验室中的肠道纤维化。在这里，我们假设通过调节水凝胶来模拟僵硬、患病的 CD 基质，我们可以使用 HIO 更好地了解纤维化如何导致肠上皮中的 CD，并找到解决或预防患者纤维化的新方法。 CD 的金标准抗炎治疗对 2/3 的患者有帮助，但不能预防或逆转纤维化。我们的目标是通过明确关注纤维化基质来确定治疗 CD 的新方法。目的和目标：我们的目标是使用 HIO 和合成水凝胶来了解 CD 肠道周围纤维化基质的硬度和成分变化如何导致疾病。通过理清这些相互作用，我们希望通过确定靶向基质来逆转或预防纤维化的方法来拓宽 CD 的治疗策略。为了实现这一目标，我们将：目标 1：确定物理信号是否在驱动 CD 样上皮表型中发挥作用。我们将使用原子力显微镜测量正常和纤维化人体肠道组织的硬度，并将 HIO 封装在模仿这些和其他物理线索的水凝胶中。然后，我们将分析封装的 HIO，以了解硬度等物理线索如何影响 HIO 上皮内的信号通路，以及水凝胶的硬度和可降解性是否可以单独促使 HIO 在自身周围形成纤维化样基质。目标 2：确定基质体线索是否发挥作用驱动 CD 样上皮表型我们将使用质谱分析法来分析正常和纤维化人类肠道组织的组成（基质体），以识别患病组织中更丰富的蛋白质。我们将 HIO 封装在水凝胶中，使用生物工程策略合并或隔离纤维化基质成分，然后分析 HIO 以确定患病基质中的特定蛋白质是否影响 HIO，或者患病基质的成分是否促使 HIO 使其局部环境变硬。目标 3：确定失调的基质是否影响上皮愈合我们将在 HIO 中诱导损伤，并使用目标 1 和 2 中的模拟硬度和基质的水凝胶来询问 CD 样硬度或基质成分是否影响肠道愈合，以及这是否是由特定信号介导的HIO 分泌的途径或蛋白质。我们还将使用我们的模型来测试针对该基质的现有药物，以确定它们是否可以促进肠道愈合。潜在的应用和益处：缓解 CD 患者炎症的金标准治疗仅使 2/3 的患者受益，并且无法逆转或预防肠道纤维化。尽管如此，大多数关于 CD 的研究都集中在缓解炎症以促进愈合。我们的研究方法旨在通过关注上皮和基质之间的相互作用，从组织水平的角度来看待 CD 和肠道愈合。在这个项目中，我们的目标是通过关注纤维化基质来发现治疗 CD 的新方法。