Curbing the Emergence of Clonal Drift in Stem Cell Expansion Culture

随着进化引导多细胞生物的发展，单细胞适应了它们的社会互动，以相互合作。然而，自然选择的残余仍然存在——适应性较高的细胞会淘汰相对较弱的邻居。细胞竞争已被证明在组织的生长和修复中起着重要作用，影响生物体的整体生存。癌细胞是超级竞争者的一个例子，它们超越公共资源，与身体的正常细胞竞争。就像癌细胞在我们的组织中作为“骗子”出现一样，生长在体外的细胞群经常被获得基因突变的异常细胞所取代，这使它们具有适应性优势。异常适合细胞的出现继续困扰着旨在获得适当和相关数量的干细胞及其衍生物用于下游应用的生物过程。多能干细胞（PSCs）具有无限增殖（自我复制）和分化为体内所有细胞类型的能力，因此功能特别强大。然而，由于其巨大的扩展潜力，psc特别容易发生突变。我的长期目标是利用细胞竞争作为对多细胞系统进行逆向工程的新视角，揭示控制单细胞之间相互作用的规则如何导致整个群体的成功或危险。我的实验室将作为一个跨学科的中心，为研究人员提供指导。通过利用计算建模、干细胞和合成生物学方面的专业知识，我们将设计一个新的管道，以更好地保持PSC批次的质量和安全性。有两个短期目标。首先，我们将应用合成生物学方法开发一个细胞“条形码”平台，用独特的DNA标识符标记psc及其后代，使我们能够根据它们的条形码来操作它们。其次，我们将跟踪每个PSC的增长动态。通过应用数学建模策略，我们将预测具有异常适应度的PSCs的出现，并使用活性条形码触发这些细胞中的自杀开关，恢复细胞批次的完整性。我打算培养一个多元化的HQP群体，包括2名研究生和5名本科生。我将意识到使用公平和包容的招聘和指导实践。最近细胞治疗领域的蓬勃发展，包括再生医学商业化中心和加拿大BlueRock治疗公司的成立，增加了生产大量细胞的需求。这项工作解决了细胞制造管道中的一个主要挑战，采用了一种新的质量控制策略来保持细胞的完整性。我们的发现将广泛适用于生物过程，包括蛋白质、病毒和生物制药生产。

Literature and Recent Progress As evolution guided the development of multicellular organisms, single cells adapted their social interactions to co-operate with one another. Nevertheless, remnants of natural selection remain - cells with higher fitness eliminate their relatively weak neighbours. Cell competition has been shown to play a significant role in the growth and repair of tissues, impacting organism survival as a whole. Cancer cells are an example of super-competitors, overtaking communal resources and out-competing the body's normal cells. Much as cancer cells emerge as "cheaters" within our tissues, cell populations grown outside the body are often overtaken by abnormal cells that acquire genetic mutations, giving them a fitness advantage. The emergence of abnormally fit cells continues to plague bioprocesses that aim to derive appropriate and relevant quantities of stem cells and their derivatives for downstream applications. Pluripotent stem cells (PSCs) are particularly powerful given their ability to indefinitely expand (make copies of themselves) and differentiate into all cell types in the body. However, owing to their great expansion potential, PSCs are particularly susceptible to mutations. Objectives and Methodology My long-term objective is to use cell competition as a new lens to reverse-engineer multicellular systems, uncovering how the rules that govern interactions between single cells lead to the success or peril of the whole population. My lab will serve as an interdisciplinary hub, providing mentorship to convergent researchers. By leveraging expertise in computational modeling, stem cells, and synthetic biology, we will design a novel pipeline to better maintain the quality and safety of PSC batches. There are two short-term objectives. First, we will apply a synthetic biology approach to develop a cellular "barcoding" platform to tag PSCs and their progeny with unique DNA identifiers, allowing us to manipulate them based on their barcode. Second, we will track the growth dynamics of each PSC as it expands. By applying a mathematical modeling strategy, we will predict the emergence of PSCs with abnormal fitness and use the active barcode to trigger a suicide switch in these cells, recovering the integrity of the cell batch. HQP I intend to train a diverse group of HQP, including 2 graduate and 5 undergraduate trainees. I will be conscious of using equitable and inclusive recruitment and mentoring practices. Impact The recent boom in the cell therapy sector, including the launch of the Centre for Commercialization of Regenerative Medicine and BlueRock Therapeutics in Canada, have increased the need to produce large quantities of cells. This work addresses a major challenge in the cell manufacturing pipeline, with a novel quality control strategy to maintain cell integrity. Our findings will be broadly applicable to bioprocesses, including protein, virus, and biopharmaceutical production.