Platforms to quantitatively interrogate cellular competition in heterogeneous tissues

挑战：将细胞群组装成功能正常的组织是生物学中一个长期存在的难题。我的研究项目的长期目标是了解控制组织组装的基本原理，并将这些原理应用于在培养皿中组装功能组织。这一具体建议侧重于了解如何在组装组织中保持稳定的组织功能。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探索这些规则：目标1：在培养皿中创建人类细胞竞争模型，并探索杀伤，生长和代谢如何随空间和时间变化。我们将使用基因工程来创造赢家和输家细胞，并使用工程技术将这些细胞组装成微片。我们将使用专门的荧光探针和显微镜技术，结合自动图像分析算法，观察细胞的死亡、生长和代谢。根据这些数据，我们将确定决定失败者杀井的关键系统参数。目标2：探索组织结构如何影响获胜细胞杀死失败细胞的速度。使用工程方法，我们将组装组织，其中赢家和输家细胞以特定的模式组织。我们将使用这些组织来探索细胞模式如何影响获胜细胞杀死失败细胞的能力和速度。目的3：比较二维和三维组织中如何控制细胞竞争。细胞的行为在二维和三维中通常是不同的。使用微流控技术，我们将创建三维微组织，其中包含赢家和输家细胞嵌入一个水凝胶。我们将比较这些三维微组织中的细胞竞争与二维薄片中的细胞竞争。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，这将改变生物学中这个爆炸性的领域。了解细胞竞争如何影响组织稳定性将为构建具有可预测结构的人工组织以及了解组织稳定性如何在衰老和疾病中破坏提供基础。此外，这个跨学科项目将招收10名学员，他们将获得批判性思维、定量分析和专业技能，有效地过渡到现代劳动力，并影响加拿大在未来几十年将面临的复杂的跨学科问题。

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.