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

挑战：控制细胞群如何聚集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，以便在培养皿中组装功能组织。我以前的DG训练了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。本建议的重点是了解组织中细胞群体的异质性。具体来说，我们将探索决定细胞竞争的规则，这是发生在发育和成人组织中的一个基本过程，在这个过程中，更适合的“赢家”细胞杀死群体中不适合的“输家”细胞。我们将通过以下目标来探索这些规则：

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.