Synthetic morphogenesis to recapitulate multicellular airway branching patterns

合成形态发生来概括多细胞气道分支模式

基本信息

批准号：
10606897
负责人：
Ian S Kinstlinger
金额：
$ 6.95万
依托单位：
BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-04-01 至 2024-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10606897
关键词：
3-Dimensional Anatomy Architecture Area Biochemical Biological Biological Models Cell Communication Cell Proliferation Cell model Cells Cellular Morphology Cellular Structures Clinical Treatment Coculture Techniques Communication Complex Comprehension Computer Models Coupled Cues Development Developmental Biology Devices Diffuse Pattern Diffusion Distal Embryo Engineering Environment Epithelium Experimental Designs Experimental Models Feedback Fractals Functional Regeneration Gene Expression Generations Goals Higher Order Chromatin Structure Human In Situ Investigation Kidney Knowledge Light Lung Lung diseases Mammary gland Maps Mathematics Measures Mesenchymal Mesenchyme Microfluidics Microscopy Modeling Morphogenesis Morphology Mus Nature Outcome Pancreas Paracrine Communication Pathway interactions Pattern Physiological Population Positioning Attribute Process Program Development Reaction Regenerative Medicine Reporter Repression Resolution Reverse engineering Scheme Signal Pathway Signal Repression Signal Transduction Stereotyping Structure Structure of parenchyma of lung Synthetic Genes Testing Therapeutic Tissue Engineering Tissues Training Trees Work biliary tract cell type cellular imaging design experience experimental study in vitro Model inducible gene expression insight intercellular communication interest knockout gene lung development lung regeneration migration morphogens network architecture novel optogenetics progenitor programs receptor regenerative regenerative therapy response self assembly small molecule spatiotemporal stem cells synthetic biology synthetic construct tool

项目摘要

Abstract The bronchial network of the human lung is a tree-like structure comprising over 20 generations of dichotomous branching; yet, the signaling basis for how this elaborate network is patterned has remained an enduring mystery. This represents not only a fundamental knowledge gap in developmental biology, but also a limiting factor for developing regenerative therapies to counter lung disease. While there are several plausible hypotheses as to how this patterning mechanism could operate, testing them has proven beyond the limits of classical gene knock- out experiments and other traditional reverse engineering approaches due to the complex signaling crosstalk found in situ. In this proposal, I will unify a classic experimental model in lung development (mesenchyme-free culture of distal lung epithelium) with state-of-the-art synthetic cell-cell signaling tools in order to map the design space for branch-patterning mechanisms. Working in a state-of-the-art Biological Design Center with a team of experts in mammalian synthetic biology and lung development, I will employ a “build-to-understand” approach wherein I construct synthetic cell populations that can either communicate with ex vivo tissues using endogenous signaling networks, or communicate with other synthetic cells using signaling pathways orthogonal to any found in nature. I will use these engineered cells to recapitulate an activation/repression feedback cycle which is thought to be vital in lung branching morphogenesis. By manipulating cell-cell communication, I will be able to isolate the fundamental design principles that govern how activation and repression signals between cells can manifest in higher- order structures. Furthermore, by decoupling specific signaling axes from their larger developmental context, and by performing high-resolution, time-lapse imaging of cell fate, I will be uniquely positioned to interrogate tissue pattern- ing mechanisms with unprecedented control. I hypothesize that reciprocal activation and repression between two cell types can give rise to a broad range of multicellular patterning outcomes depending on additional feedback loops and initial conditions. To test this hypothesis, I will explore the how the morphology and topol- ogy of multicellular patterns can be tuned by manipulating the signaling interactions between them. My overarching hypothesis is based on the predictions of previous computational models of branching morphogenesis via reaction- diffusion patterning, so I will use those predictions, and this theoretical framework, to guide my experimental designs. To assess whether synthetic signaling by engineered cells could also be a tractable approach for generating regen- erative lung tissue, I will further interrogate a 3D in vitro model where cell-cell signaling occurs exclusively through synthetic morphogens and receptors. Taken together, these studies will provide fundamental insights into how complex anatomical structures can be encoded in relatively simple signaling schemes which are executed locally between cells. Analysis of the resulting branch patterns is also expected to inspire a new paradigm for har- nessing synthetic cell-cell signaling to guide and direct the morphogenesis of therapeutically relevant cell types into tissue-specific architectures for regenerative medicine.

抽象的人肺的支气管网络是一种类似树状的结构，完成了20代二分法分枝;然而，该精心制作网络的图案如何保持信号基础仍然是一个持久的谜团。这不仅代表了发育生物学的基本知识差距，而且代表了一个限制因素用于开发再生疗法以对抗肺部疾病。虽然有几种合理的假设对于这种模式机制如何运作，对它们进行测试已被证明超出了经典基因敲除的限制。由于在原位。在此提案中，我将统一肺发育中的经典实验模型（无间隙培养带有最先进的合成细胞 - 细胞信号传导工具的远端肺上皮）以绘制设计分支机构机制的空间。在一个最先进的生物设计中心工作哺乳动物合成生物学和肺发展专家，我将采用一种“建立理解”方法我构建了可以使用内源信号传导与离体组织进行通信的合成细胞群网络，或使用与自然中发现的任何信号通路正交的信号通路与其他合成细胞进行通信。我将使用这些工程细胞来概括激活/抑制反馈周期，这在肺分支形态发生。通过操纵细胞电池通信，我将能够隔离基本设计原理，这些原理如何在较高的订单结构。此外，通过将特定的信号轴从其较大的发育环境中分离出来，并通过对细胞命运进行高分辨率，延时成像，我将被唯一地审问组织模式 - 具有前所未有的控制的机制。我假设这种相互激活和表达两种细胞类型会导致广泛的多细胞图案结果，具体取决于附加反馈循环和初始条件。为了检验这一假设，我将探讨形态和topology和topol- 可以通过操纵它们之间的信号传导相互作用来调整多细胞图案的OGY。我的总体假设是基于通过反应的先前计算模型的预测扩散图案，因此我将使用这些预测以及这个理论框架来指导我的实验设计。评估工程细胞的合成信号是否也可能是产生再生的一种可进行的方法 erration的肺组织，我将进一步询问一个3D体外模型，其中细胞 - 细胞信号仅通过合成形态和接收器。综上所述，这些研究将提供有关如何复杂的解剖结构可以用相对简单的信号方案进行编码在细胞之间局部。还有望分析由此产生的分支模式，以激发新的范式嵌套合成细胞 - 细胞信号传导，指导和将热相关细胞类型的形态发生到用于再生医学的组织特异性体系结构。