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个树状结构，由20多代二叉树组成然而，这个复杂的网络如何形成的信号基础仍然是一个持久的谜。这不仅代表了发育生物学的基础知识差距，也是一个限制因素开发再生疗法来对抗肺部疾病虽然有几个看似合理的假设。对于这种模式机制如何运作，测试已证明超出了经典基因敲除的限制由于复杂的信号串扰，我们无法进行实验和其他传统的逆向工程方法在这个提案中，我将统一肺发育的经典实验模型（无间充质培养）。远端肺上皮）与最先进的合成细胞-细胞信号传导工具，以绘制设计图与团队一起在最先进的生物设计中心工作。作为哺乳动物合成生物学和肺部发育方面的专家，我将采用“构建理解”的方法。我构建了可以使用内源信号与离体组织进行通信的合成细胞群网络，或使用与自然界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。将使用这些工程细胞来重现激活/抑制反馈循环，这被认为是至关重要的通过操纵细胞间的通讯，我将能够分离出基本的机制。控制细胞之间的激活和抑制信号如何在更高层次上体现的设计原则此外，通过将特定的信号轴与其更大的发育背景分离，并通过对细胞命运进行高分辨率、延时成像，我将处于独特的位置来询问组织模式- 具有前所未有的控制能力的机制。两种细胞类型可以产生广泛的多细胞模式结果，具体取决于额外的条件为了测试这个假设，我将探索形态和拓扑结构。多细胞模式的系统可以通过操纵它们之间的信号相互作用来调整。该假设基于先前通过反应进行分支形态发生的计算模型的预测扩散图案，所以我将使用这些预测和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以成为产生再生的易于处理的方法对于活动性肺组织，我将进一步研究 3D 体外模型，其中细胞间信号传导仅通过综上所述，这些研究将提供关于如何合成形态发生素和受体的基本见解。复杂的解剖结构可以用相对简单的信号方案进行编码并执行对由此产生的分支模式的分析也有望激发一种新的模式。合成细胞间信号传导以指导治疗相关细胞类型的形态发生再生医学的组织特异性架构。