Design strategies for programming and quantitatively interrogating cellular competition in heterogeneous tissues

异质组织中编程和定量询问细胞竞争的设计策略

• 批准号: RGPIN-2019-06486
• 负责人: McGuigan, Alison
• 金额: $ 3.06万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=714605
• 关键词: Design; strategies; programming; quantitatively; interrogating

CHALLENGE:The rules that govern how populations of cells assemble into functional tissues are a long-standing puzzle. The long-term objective of my research program is to understand these rules to enable the assembly of functional tissues in a dish. My previous DG trained 30 HQP and explored how the properties of different cell populations impact overall tissue behaviour. This proposal focuses on understanding cell population heterogeneity in a tissue. Specifically, we will explore the rules that dictate cell competition, a fundamental process that occurs in both developing and adult tissue, in which more fit “winner” cells kill less fit “loser” cells in the population. We will explore these rules via the follow aims: Aim 1: Create 2-dimensional in vitro and in silico models of mammalian cell competition to quantify the spatial and temporal dynamics of loser cell killing and winner cell growth We will engineer monolayers containing domains of winner and loser cells. Using fluorescent cell death and growth reporters, combined with automated image tracking algorithms we will quantify loser cell killing rate and which cell and tissue properties affect this rate. Using our experimental data, we will develop a computational model to predict loser killing rate. Aim 2: Explore the relationship between competition dynamics and tissue heterogeneity We will A)Quantify kill rates in monolayers with specific winner/loser domain geometries or grown on microgrooves oriented to induce morphological polarization of the cells either parallel or perpendicular to the winner-loser domain interface to test the hypothesis that tissue structure impacts competition and B)Monitor winner expansion in monolayers engineered with fluorescence markers to enable winner cells proximal and distant to the winner-loser cell interface to be distinguished to test the hypothesis that competition is associated with a cost. Aim 3: Explore the impact of metabolic gradients on competition dynamics and heterogeneity in 3-dimensions (3D) We will use our unique 3D TRACER system to test the hypothesis that gradients of nutrient and waste factors will impact metabolic signalling in winner and loser populations impacting their relative fitness and ability to compete. NOVELTY/IMPACT: By applying unique engineering tools, the proposed research will interrogate the critical parameters that control cell competition in an entirely new way that will transform this exploding field. Understanding the relationship between competition and tissue heterogeneity will provide a basis for engineering self-organizing tissues with predictable structures and for understanding how competition drives tissue break down in aging and disease. In addition, the program will engage 18 HQP who will gain the critical thinking, quantitative analysis and professional skills essential to effectively transition into a modern workforce and impact the complex inter-disciplinary problems Canada will face in the coming decades.

挑战：控制细胞群如何组装成功能组织的规则是一个长期存在的难题。我的研究计划的长期目标是了解这些规则，以便能够在培养皿中组装功能组织。我之前的 DG 培训了 30 名 HQP，并探索了不同细胞群的特性如何影响整体组织行为。该提案的重点是了解组织中细胞群的异质性。具体来说，我们将探索决定细胞竞争的规则，细胞竞争是发育中和成体组织中发生的基本过程，其中更适合的“获胜者”细胞杀死群体中不太适合的“失败者”细胞。我们将通过以下目标探索这些规则： 目标 1：创建哺乳动物细胞竞争的二维体外和计算机模型，以量化失败者细胞杀伤和获胜者细胞生长的空间和时间动态 我们将设计包含获胜者和失败者细胞域的单层。使用荧光细胞死亡和生长记者，结合自动图像跟踪算法，我们将量化失败者细胞的杀伤率以及哪些细胞和组织特性影响该率。利用我们的实验数据，我们将开发一个计算模型来预测失败者杀戮率。 目标2：探索竞争动态与组织异质性之间的关系 我们将 A) 量化具有特定赢家/输家域几何形状的单层细胞的杀灭率，或者在微凹槽上生长，以诱导平行或垂直于赢家-输家域界面的细胞形态极化，以测试组织结构影响竞争的假设；B) 监测用荧光标记设计的单层细胞中的获胜者扩张，使获胜者细胞能够接近和远离 区分赢家与输家的单元界面，以检验竞争与成本相关的假设。 目标 3：探索代谢梯度对三维 (3D) 竞争动态和异质性的影响 我们将使用我们独特的 3D TRACER 系统来测试以下假设：营养和废物因子的梯度将影响获胜者和失败者群体的代谢信号，从而影响他们的相对健康度和竞争能力。 新颖性/影响力：通过应用独特的工程工具，拟议的研究将以一种全新的方式询问控制细胞竞争的关键参数，这将改变这个爆炸性的领域。了解竞争与组织异质性之间的关系将为设计具有可预测结构的自组织组织以及了解竞争如何驱动组织在衰老和疾病中分解提供基础。此外，该计划还将吸引 18 名 HQP，他们将获得批判性思维、定量分析和专业技能，以有效过渡到现代劳动力队伍，并影响加拿大在未来几十年将面临的复杂的跨学科问题。