Harnessing cell-mediated matrix remodelling by human iPSC-derived intestinal organoids within 3D materials to understand epithelial-mesenchymal intera

博士项目：目标和预防人类肠道可以在体外使用Matrigel（肿瘤衍生的3D基质）中培养的类器官进行建模。然而，Matrigel的物理性质，如其硬度和降解性不能被调节，这限制了我们理解基质和基质重塑如何影响肠上皮的能力（反之亦然）。Gentleman实验室最近发现，封装在具有特定物理特性的水凝胶中的干细胞通过局部分泌基质蛋白和降解周围环境来重塑周围环境，这影响了它们的命运（Ferreira，Nat Commun 2018）.这里，我们将通过将绅士实验室（KCL）在合成3D矩阵方面的专长与斯塔格（QMUL）的专长相结合，来利用这一发现在肠道炎症和NevesLab（KCL）的肠道类器官模型的兴趣，开发人类类器官模型IBD定义，合成，可修改的3D环境中，我们将用来问基本的问题，在基质如何影响上皮（反之亦然）在健康和疾病。为了实现这一目标，我们将创造人类诱导多能干细胞（iPSC）衍生的肠道类器官（hIO），并将它们封装在基于PEG的合成水凝胶中，我们可以独立地调节刚度，配体密度和降解性。使用敲除/过表达方法、微流变学技术和具有受控软化/硬化的水凝胶，我们将机械地探测细胞介导的基质分泌/降解和/或改变的局部机械性质的组合如何影响类器官表型。这将使我们能够确定潜在的基质targetswithin肠壁，可以利用治疗。通过调节合成水凝胶的物理性质，建立并表征人iPSC衍生的类器官IBD模型.了解使用敲低和过表达技术调节类器官周围的局部基质重塑对上皮细胞和间充质细胞的影响.了解调节类器官周围的局部基质机械特性如何影响上皮细胞和间充质细胞。

PhD project: aims and descriptionThe human intestine can be modelled in vitro using organoids cultured within Matrigel, atumour-derived 3D matrix. However, Matrigel's physical properties, such as its stiffness anddegradability cannot be tuned, which limits our ability to understand how the matrix andmatrix remodeling impacts the intestinal epithelium (and vice versa). The Gentleman Lab hasrecently discovered that stem cells encapsulated within hydrogels with defined physicalproperties remodel their surroundings by secreting matrix proteins locally and by degradingtheir surrounding environment, which impacts their fate (Ferreira, Nat Commun 2018).Here, we will exploit this discovery by melding the Gentleman Lab's (KCL) expertise insynthetic 3D matrices with the Stagg's (QMUL) interests in gut inflammation and the NevesLab's (KCL) organoid-based models of the intestine to develop human organoid-based modelsof IBD within defined, synthetic, modifiable 3D environments, which we will use to askfundamental questions in how the matrix impacts the epithelium (and vice versa) in healthand disease. To accomplish this, we will create human induced pluripotent stem cell (iPSC)-derived intestinal organoids (hIO) and encapsulate them within PEG-based synthetichydrogels in which we can independently modulate stiffness, ligand density anddegradability. Using knockdown/overexpression approaches, microrheology techniques, andhydrogels with controlled softening/stiffening, we will then mechanistically probe how acombination of cell-mediated matrix secretion/degradation and/or altered local mechanicalproperties impact organoid phenotype. This will allow us to identify potential matrix targetswithin the intestinal wall that could be exploited therapeutically.Specific Aims:1. Establish and characterize a human iPSC-derived organoid-based model of IBD bymodulating synthetic hydrogels' physical properties.2. Understand the impact on epithelial and mesenchymal cells of modulating local matrixremodelling around organoids using knockdown and overexpression techniques.3. Understand how modulating local matrix mechanical properties around organoids impactsepithelial and mesenchymal cells.