"Bottom - Up" Profiling of Interacting Cellular Systems

 描述（由申请人提供）：我们由数万亿个相互作用的细胞组成，但我们对它们如何共同工作的理解是有限的。这是因为我们传统上将生物体从“自上而下”划分为广泛的细胞类型或迭代细化的“同质”子集，然后分别研究每个这样的群体。然而，最近的研究表明，即使是“相同”的细胞也可以表现出功能上的重要差异，并且细胞行为受到微环境和细胞相互作用的强烈影响。举例来说，对于 免疫树突状细胞（DC）和T细胞-共同负责识别病原体和诱导适应性免疫应答-细胞亚型、信号环境和物理接触都影响促炎和调节应答之间的平衡。不幸的是，我们无法在复杂系统的背景下彻底测量和分析这些影响中的每一个，这限制了我们掌握如何实现适当免疫功能的能力。我将利用纳米技术和分子生物学的最新进展，开发广泛适用的平台，用于操纵和分析许多相互作用的单细胞， 可以揭示它们如何从“自底向上”共同执行系统级行为。这些将包括用于分离培养和监测细胞的方法，以及作为受控的整体，进行靶向操作，整合不同的分子测量（例如，RNA和蛋白质），并在体外和原位检查许多单细胞中的基因组RNA谱。虽然广泛适用，但我将专门利用这些技术来研究DC和T细胞如何协同对抗病原体，以及T细胞如何预防可驱动自身免疫的不受控制的DC炎症。总的来说，我的工作将有助于识别细胞参与者和他们用来执行系统级行为的策略，将增强我们对细胞反应，通信，疾病和治疗的理解，并将产生变革性的新技术，用于全面和可控地分析许多不同的生物系统。

 DESCRIPTION (provided by applicant): We consist of trillions of interacting cells, but our understanding of how they work together is limited. This is because we have traditionally divided organisms from the "top-down" into broad cell types or iteratively-refined "homogeneous" subsets and then studied each such population separately. Yet recent studies have shown that even "identical" cells can exhibit functionally important differences and that cellular behaviors are strongly influenced by both the microenvironment and cellular interactions. Illustratively, for immune dendritic cells (DCs) and T cells - collectively responsible for recognizing pathogens and inducing adaptive immune responses - cellular subtype, signaling milieu, and physical contacts all impact the balance between proinflammatory and regulatory responses. Unfortunately, our inability to thoroughly measure and analyze each of these influences within the context of a complex system has limited our ability to grasp how proper immune function is achieved. I will leverage recent advances in nanotechnology and molecular biology to develop broadly applicable platforms for manipulating and profiling many interacting single cells so that I can uncover how they collectively perform systems-level behaviors from the "bottom-up". These will include methods for culturing and monitoring cells in isolation and as a controlled ensemble, performing targeted manipulations, integrating different molecular measures (e.g., RNA and protein), and examining genomic RNA profiles in many single cells in-vitro and in-situ. Although broadly applicable, I will specifically utilize these technologies to examine how DCs and T cells synergistically fight pathogens and how T cells prevent uncontrolled DC inflammation that can drive autoimmunity. Collectively, my work will help identify the cellular players and the strategie they use to execute systems-level behaviors, will enhance our understanding of cellular response, communication, disease, and therapeutics, and will yield transformative new technologies for comprehensively and controllably profiling many different biological systems.