A reductionist organoid-based model to study how matrix remodelling and soluble signals impact cell fate decisions in human pancreas development

基于还原论类器官的模型，用于研究基质重塑和可溶性信号如何影响人类胰腺发育中的细胞命运决定

• 批准号: 2578185
• 金额: --
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2021
• 资助国家: 英国
• 起止时间: 2021 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2578185
• 关键词: reductionist; organoid; based; model; study

The interplay between the pancreatic epithelium, its surrounding extracellular matrix, andcell-derived chemical cues are pivotal for pancreas formation and beta cell differentiation.Organoids allow for molecular analyses of complex biological phenomena; however, there arevery few tools available to visualise, modulate or probe matrix/chemical cues and understandtheir impact on cellular responses.iPSC-derived pancreas progenitor organoids (iPPO) can be expanded in Matrigel as itsexternal signals (chemical and mechanical) are sufficient to support iPPO prior to theirdifferentiation to beta cells. We have developed fully synthetic PEG-based hydrogels that cansimilarly support iPPOs.Here, we aim to use PEG-based hydrogels to understand the fundamental molecular, solubleand physical cues required to drive iPSCs differentiation to beta cells that could be used as atherapy in diabetes. We will pursue the following aims:1. Map matrix remodelling around iPPOs.We have established an "embryonic pancreas development" model whereby iPPOs areencapsulated within synthetic hydrogels. We will characterize matrix remodelling aroundorganoids using AFM force spectroscopy to measure peri-organoid stiffness, and multipleparticle tracking microrheology to monitor local degradation.2. Model the diffusion of soluble signals from iPPOs in static and dynamic hydrogels thatmimic peri-organoid stiffness changes. We will use COMSOL to simulate how key solublesignals between the epithelium and mesenchyme are affected by cell-mediated matrixremodelling around organoids. Models will be validated with experimental work usinghydrogels that controllably soften (hydrolysis) or stiffen (secondary radical cross-linking) tomatch dynamics observed in Aim 1.3. Harness the platform to identify the role of specific matrix cues in iPPOs differentiation. Wewill culture iPPOs in softening/stiffening hydrogels to determine if mechanical modulation issufficient to induce differentiation into beta cells. We will also inhibit/overexpress candidatematrix-modulating proteins, allowing us to relate specific mechanistic cues within the matrixto cell fate.

胰腺上皮，其周围的细胞外基质，和细胞衍生的化学线索之间的相互作用是胰腺形成的关键和β细胞分化的关键。类orniations可以进行复杂生物学现象的分子分析。但是，很少有工具可以可视化，调节或探测矩阵/化学提示，并且可以理解对细胞反应的影响。IPSC衍生的胰腺祖细胞器官（IPPO）可以在Matrigel中扩展，因为ITSEXENTER信号（化学和机械）足以支撑IPPO在其效率上对BetA细胞之前的IPPO支持。我们已经开发了完全合成的基于合成的PEG水凝胶，可以支持IPPOS。我们的目的是使用基于PEG的水凝胶来了解将IPSC分化与可以用作糖尿病中动脉粥样硬化的Beta细胞所需的基本分子，可露天的物理提示。我们将追求以下目标：1。我们建立了一个“胚胎胰腺发展”模型，在合成水凝胶中囊括了IPPOS的“胚胎胰腺发展”模型。我们将使用AFM力光谱法表征矩阵重塑，以测量甲壳周围的刚度，以及多粒子跟踪微流变学来监测局部降解。2。建模可溶性信号从IPPOS中的扩散在静态和动态水凝胶中，以使甲壳周围的刚度发生变化。我们将使用comsol来模拟上皮和间质之间的关键溶性如何受到细胞介导的细胞介导的细胞介导的细胞围绕类器官的影响。模型将通过使用液凝胶来验证模型，这些凝胶可以控制在AIM 1.3中观察到的（水解）或硬化（次级自由基交叉链接）的tomatch动力学。利用平台来确定特定矩阵提示在IPPO分化中的作用。 Wewill培养IPPO在软化/僵硬的水凝胶中，以确定机械调节是否足以诱导分化为β细胞。我们还将抑制/过表达念珠菌调节蛋白，从而使我们能够在基质细胞命运中关联特定的机械线索。