Platforms to quantitatively interrogate cellular competition in heterogeneous tissues

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CHALLENGE: Assembling populations of cells into tissues that function properly is a long-standing puzzle in biology. The long-term objective of my research program is to understand the fundamental principles that control tissue assembly and apply these to assemble functional tissues in a dish. This specific proposal focuses on understanding how to maintain stable tissue function in an assembled tissue. Specifically, we will explore the rules that dictate cell competition, a fundamental process that occurs in both developing and adult tissues, 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 models of human cell competition in a dish and explore how killing, growth and metabolism vary in space and over time. We will use genetic engineering to create winner and loser cells and engineering techniques to assemble these cells into microsheets. We will observe cell death, growth and metabolism using specialized fluorescent probes and microscopy techniques, combined with automated image analysis algorithms. From this data, we will identify the critical system parameters that determine loser killing. Aim 2: Explore how the organization of a tissue influences the rate at which winner cells kill loser cells. Using engineering methods, we will assemble tissues in which winner and loser cells are organized in specific patterns. We will use these tissues to explore how cell patterning influences the capability and speed at which winner cells kill loser cells. Aim 3: Compare how cell competition is controlled in 2 versus 3 dimensional tissues. Cell behavior is typically different in 2 versus 3 dimensions. Using microfluidic techniques, we will create 3 dimensional microtissues containing winner and loser cells embedded in a a hydrogel. We will compare cell competition in these 3 dimensional microtissues to cell competition in 2 dimensional sheets. 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 in biology. Understanding how cell competition impacts tissue stability will provide a basis for engineering artificial tissues with predictable structures and for understanding how tissue stability breaks down in aging and disease. In addition, this inter-disciplinary program will engage 10 trainees 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.