BRIGE: Modeling metabolism in embryonic stem cell growth and differentiation

本课题的研究目的是对胚胎干细胞在增殖和分化过程中的代谢进行测量和建模。PI的小组致力于通过研究和模拟代谢物浓度来理解和控制新陈代谢。在这里，我们应用代谢物分析和机器学习技术来创建胚胎干细胞分化代谢的第一个描述性模型。我们还将研究我们认为与细胞分化显著相关的代谢物的调节潜力。干细胞将对现代医学产生革命性的影响。鉴于最近的成功，人们的注意力开始转向发展干细胞治疗所固有的迫在眉睫的生物制造问题。从少量扩大干细胞群体和控制其分化的困难是扩大规模的重大障碍，干细胞治疗可能在技术上或经济上都不可行。在放大过程中控制干细胞行为的一个有希望的途径是监测和操纵这些细胞的代谢和代谢信号，但这方面的研究很少。这一变革性的研究计划将有助于填补这一知识空白，特别是解决干细胞培养和工程领域的三个关键问题。通过开展开创性的纵向代谢组学研究，我们将(1)建立一个初始数据集和模型，证明代谢在干细胞扩增和分化中的重要性，并可用于指导培养规模的扩大。通过鉴定能够促进特定分化谱系的代谢物，我们将(2)能够在胚胎干细胞扩增和分化过程中更精确地控制胚胎干细胞，并促进规模化。最后，我们的细胞外代谢物分析技术和模型将(3)为干细胞培养监测和质量控制提供非侵入性、非破坏性的方法，能够在形态变化明显之前很久就检测到细胞状态的变化。更广泛的影响通过利用我们的系统级代谢分析来扩展控制干细胞命运的能力，我们将能够开发出新的和更复杂的干细胞工程疗法，这些疗法可以挽救或极大地改善面临衰弱性疾病的人们的生命。将我们的研究成果直接应用于干细胞培养技术的工业化生产水平，将绕过该领域的一个关键经济障碍，使治疗候选药物的开发成为不仅可以治愈那些富有或幸运的人，而且可以治愈任何面临这种疾病的人，无论社会经济地位如何。这样的系统级分析也有可能对未来的工程师产生重大影响；本着这一精神，PI提出了一个扩大参与计划，主要重点是鼓励女性参与工程。这项工作的核心是为女童子军开发一项活动和事件，以激发他们对工程的兴趣，并向他们介绍定义工程的系统级思维方式。PI将与一名当地教师合作开展这项活动，并使其与佐治亚州的教育标准保持一致。女本科生和研究生将帮助发展和实施这项活动。PI计划的其他主要活动包括继续与全女性宿舍合作，与校园女工程师协会分会合作开展外展活动，以及招募和指导女学生。此外，PI的小组将通过REU计划接待未被充分代表的少数民族本科生；制定了第一个和第三个目标，以便轻松地将本科研究人员整合到一个有回报的项目中。

PI: Styczynski, MarkProposal Number: 1125684The research objective of this proposal is to measure and model the metabolism of embryonic stem cells during proliferation and differentiation. The PI's group is dedicated to understanding and controlling metabolism by studying and modeling metabolite concentrations. Here, we apply metabolite profiling and machine-learning techniques to create the first-ever descriptive models of metabolism in embryonic stem cell differentiation. We will also study the regulatory potential of metabolites we identify as significantly correlated with cellular differentiation.Intellectual MeritStem cells are poised to make a revolutionary impact on modern medicine. Given recent successes, attention has begun to turn to the looming biomanufacturing problem inherent in developing stem cell treatments. Difficulties expanding stem cell populations from small numbers and controlling their differentiation are such significant roadblocks to scale-up that stem cell therapies may be technically or economically infeasible on a large scale. One promising route for controlling stem cell behavior during scale-up is monitoring and manipulating metabolism and metabolic signals in these cells -yet there has been little research in this area. This transformative research plan will help fill this knowledge gap, specifically addressing three key issues in the field of stem cell culture and engineering. By performing a pioneering longitudinal metabolomics study, we will (1) establish an initial dataset and models demonstrating the importance of metabolism in stem cell expansion and differentiation, and that can be used in guiding culture scale-up. By identifying metabolites capable of promoting specific differentiation lineages, we will (2) enable more precise control of ES cells during expansion and differentiation, also facilitating scale-up. Finally, our extracellular metabolite profiling techniques and models will (3) provide non-invasive, non-destructive methods for stem cell culture monitoring and quality control that are capable of detecting changes in cell state long before morphological changes are evident.Broader ImpactsBy expanding capabilities in controlling stem cell fate using our systems-level analysis of metabolism, we will enable development of novel and more complex stem cell engineering therapeutics that can save or greatly improve the lives of people facing debilitating diseases. The direct application of our research results to the scale-up of stem cell culture technologies to industrial production levels will circumvent a critical economic roadblock in the field, enabling the development of therapeutic candidates into products that can heal not just those who are rich or fortunate, but anyone facing such a disease regardless of socioeconomic status. Such systems level analysis also has the potential to make a significant impact on future engineers; in this vein, the PI proposes a broadening participation program with a primary focus of encouraging female participation in engineering. The centerpiece of this effort is the development of an activity and event for Girl Scout troops that stokes interest in engineering and introduces them to the systems-level mindset that defines engineering. The PI will collaborate with a local teacher to develop this activity and align it with Georgia state educational standards. Female undergraduates and graduate students will help develop and implement this activity. Other key activities planned by the PI include continuing work with an all-female dormitory, collaboration with the campus Society of Women Engineers chapter in developing outreach activities, and recruiting and mentoring female students. Additionally, the PI's group will host underrepresented minority undergraduate students through an REU program; the first and third aims have been formulated to easily integrate undergraduate researchers in a rewarding project.