Curbing the Emergence of Clonal Drift in Stem Cell Expansion Culture

抑制干细胞扩增培养中克隆漂移的出现

• 批准号: RGPIN-2020-04198
• 负责人: Shakiba, Nika
• 金额: $ 2.7万
• 依托单位: University of British Columbia
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=746210
• 关键词: Curbing; Emergence; Clonal; Drift; Stem

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.

随着进化的发展，文献和最近的进步指导了多细胞生物的发展，单细胞将其社交相互作用调整为彼此合作。然而，自然选择的残留物仍然存在 - 具有较高适应性的细胞消除了相对较弱的邻居。细胞竞争已被证明在组织的生长和修复中起着重要作用，从而影响了整个生物的生存。癌细胞是超级竞争者，超过公共资源并超越人体正常细胞的一个例子。就像癌细胞成为我们组织中的“作弊者”一样，体外生长的细胞种群通常被获得遗传突变的异常细胞所取代，从而使它们具有适应性的优势。异常拟合细胞的出现继续困扰着旨在为下游应用提供适当量且相关的干细胞及其衍生物。鉴于它们无限期扩展（复制自身）并分化为体内所有细胞类型的能力，多能干细胞（PSC）特别强大。但是，由于其巨大的扩展潜力，PSC特别容易受到突变的影响。目的和方法论我的长期目标是将细胞竞争用作新的镜头来逆转工程多细胞系统，从而发现控制单细胞之间相互作用的规则如何导致整个人群的成功或危险。我的实验室将作为跨学科枢纽，为融合研究人员提供指导。通过利用计算建模，干细胞和合成生物学方面的专业知识，我们将设计一种新型管道，以更好地维持PSC批次的质量和安全性。有两个短期目标。首先，我们将采用一种合成生物学方法来开发细胞“条形码”平台，以使用独特的DNA标识符标记PSC及其后代，从而使我们可以根据其条形码操纵它们。其次，我们将在每个PSC扩展时跟踪每个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性的PSC的出现，并使用主动条形码触发这些细胞中的自杀开关，从而恢复细胞批处理的完整性。 HQP我打算培训一组HQP，其中包括2名毕业生和5名本科学员。我将意识到使用公平和包容的招聘和指导实践。影响细胞治疗部门最近的繁荣，包括在加拿大加拿大再生医学商业化和Bluerock Therapeutics的商业化中心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主要挑战，并具有保持细胞完整性的新型质量控制策略。我们的发现将广泛适用于生物过程，包括蛋白质，病毒和生物药物生产。