CAREER: Dissecting the Biological Impact of Stem Cell Fusion; a Unified Plan for Stem Cell Discovery via Accessible Engineering

该奖项是根据2009年美国复苏和再投资法案（公法111-5）0844537 OgleThis职业生涯项目是建立在PI的最终研究愿景，开发可访问的技术，以解决限制干细胞发现的重大挑战。 解决的重大挑战是解开干细胞与成熟细胞融合的机制以及这种事件的影响（有益或有害）。 干细胞与成熟细胞的融合是一个受调控的过程，能够促进细胞存活和分化，因此对于组织的发育、修复甚至疾病的发病机制都是至关重要的。 考虑到潜在的影响，令人惊讶的是，干细胞融合的基本方面是未知的，包括触发融合的信号，负责介导融合的细胞表面蛋白，重编程的机制和融合后杂交细胞的功能能力。 阻碍这一领域发现的是缺乏适当的技术来1）识别准备融合的融合伙伴，2）跟踪融合产物（即，杂交细胞），和3）以高通量方式分析和分选融合产物。 PI最近开发了在细胞融合后立即准确鉴定融合产物并随时间推移在培养物和活生物体中跟踪融合产物的技术。 在这里，PI将开发和表征一种新的流体平台，该平台与多光子显微镜系统耦合，用于融合产物等大型实体的高通量流式细胞术分析（目标1）。 该系统将与先前开发的技术一起使用，以检查干细胞-成熟细胞融合对杂交细胞功能的影响（目标2）。 这项新的工作阐明了技术如何与基础和临床科学适当结合，将改变干细胞的特征，分析和刺激方式。 目标1和2的实施将提供一个模板，以促进对干细胞技术的发现和理解，同时促进广泛受众的教学、培训和学习（目标3）。 尽管对干细胞发现的广泛热情，但对干细胞细胞过程的基本机制的理解（即，增殖、粘附、分化）受到古老分析方法的限制。 研究生已被确定为融合技术与基础干细胞科学，并将这些知识传达给全国学术界和工业界的理想渠道。 研究生将接受多模式培训，以促进对与干细胞发现相关的关键工程概念的理解，并促进有效的教学和学习实践。 接受此培训的研究生将有机会通过规划和执行外展计划（夏季科学营，WiCell研究所）来传播他们的新知识，在专业社区研讨会（UW干细胞技术合作实验室，由PI共同创立）的背景下应用他们的新知识，并通过指导代表性不足的本科生来传播和应用他们的新知识。

This award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5)0844537OgleThis CAREER project is built on the PI's ultimate research vision to develop accessible technologies to address significant challenges limiting stem cell discovery. The significant challenge addressed is unraveling the mechanism(s) by which stem cells fuse with mature cells and the impact (beneficial or detrimental) of such an event. Fusion of stem cells with mature cells is a regulated process capable of promoting cell survival and differentiation and thus could be of central importance for development, repair of tissues and even the pathogenesis of disease. Given the potential impact, it is surprising that basic aspects of stem cell fusion are unknown including the signals that trigger fusion, the cell surface proteins responsible for mediating fusion, the mechanism of reprogramming and the functional capacity of hybrid cells after fusion. Hindering discovery in this area is lack of appropriate technology to 1) identify fusion partners poised to fuse, 2) track fusion products (i.e., hybrid cells) over time and 3) analyze and sort fusion products in a high-throughput manner. The PI has recently developed technologies to accurately identify fusion products immediately after cell fusion and to track fusion products over time both in culture and in living organisms. Here the PI will develop and characterize a new fluidics platform coupled to a multiphoton microscopy system for high-throughput, flow cytometric analysis of large entities such as fusion products (Objective 1). This system will be used together with previously developed technologies to examine the impact of stem cell-mature cell fusion on hybrid cell function (Objective 2). This novel work exemplifies how technology, when appropriately coupled to basic and clinical science, will transform the way stem cells are characterized, analyzed and stimulated. Execution of Objectives 1 and 2 will provide a template to advance the discovery and understanding of stem cell technologies while promoting teaching, training and learning of a wide range of audiences (Objective 3). Despite widespread enthusiasm for stem cell discovery, understanding of the basic mechanisms of the cellular processes of stem cells (i.e., proliferation, adhesion, differentiation) has been limited by archaic analysis methods. Graduate students have been identified as the ideal conduit for merging technology with basic stem cell science and for conveying this knowledge to academic and industrial communities nation-wide. Graduate students will undergo multimodal training to foster an understanding of key engineering concepts related to stem cell discovery and to promote effective teaching and learning practices. Graduate students who undergo this training will have an opportunity to disseminate their new knowledge by planning and executing an outreach program (Summer Science Camp, WiCell Research Institute), to apply their new knowledge in the context of a professional community workshop (UW Stem Cell Technology Collaboratory, cofounded by the PI), and to disseminate and apply their new knowledge via mentoring of underrepresented undergraduate students.