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，使功能组织在培养皿中组装。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。该提案侧重于了解组织中的细胞群体异质性。具体来说，我们将探索决定细胞竞争的规则，这是一个发生在发育和成年组织中的基本过程，在这个过程中，更适合的“赢家”细胞杀死种群中不太适合的“输家”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赢家细胞生长的空间和时间动态 我们将工程单层包含赢家和输家细胞域。使用荧光细胞死亡和生长报告基因，结合自动图像跟踪算法，我们将量化失败细胞的杀伤率，以及哪些细胞和组织特性会影响这一比率。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的杀戮率。 目的2：探索竞争动力学与组织异质性之间的关系 我们将A）量化具有特定赢家/输家结构域几何形状的单层中的杀伤率，或生长在定向为诱导细胞的形态极化的微槽上，所述细胞平行于或垂直于赢家-输家结构域界面，以测试组织结构影响竞争的假设，和B）监测用荧光标记物工程化的单层中的赢家扩增，以使赢家细胞接近和远离赢家。失败者单元格界面被区分，以测试竞争与成本相关的假设。 目的3：探索代谢梯度对竞争动力学和三维异质性的影响 我们将使用我们独特的3D TRACER系统来测试这一假设，即营养和废物因素的梯度将影响赢家和输家群体的代谢信号，从而影响他们的相对健康和竞争能力。 新奇/影响：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，这将改变这一爆炸性领域。了解竞争和组织异质性之间的关系将为工程化具有可预测结构的自组织组织提供基础，并了解竞争如何驱动衰老和疾病中的组织分解。此外，该计划将聘请18 HQP谁将获得批判性思维，定量分析和专业技能，有效地过渡到一个现代化的劳动力和影响复杂的跨学科问题加拿大将在未来几十年面临的必要。