Live-cell Activity Architecture in Cancer

 描述（由申请人提供）：蛋白质纳米机器的组装/拆卸和酶活性是所有细胞功能的基础，而失调的纳米机器是癌症的最终罪魁祸首。我们提出的研究旨在建立一个新的概念框架，以具体了解分子活动的细胞组织。我们假设，细胞生化活动的空间组织成一个“活动架构”，通过特定的组织活性分子和他们的监管合作伙伴。这 活性结构与细胞的结构和机械结构一起编码了驱动细胞功能所需的所有信息。我们进一步假设，即使是一些失调的驱动分子对这种活性结构的扰动，也可能导致对细胞功能的有害影响，例如失去对细胞生长、分裂和死亡的控制。 我们的研究计划的关键技术创新是开发和利用新一代的使能技术，用于可视化和扰动活细胞的原生环境中的生物化学和生物物理过程。通过这些新技术，我们将开始通过阐明蛋白激酶的酶活性的空间组织来测试我们的一般假设，蛋白激酶是在正常细胞生理学和肿瘤发生中起关键作用的酶家族。我们将进一步探讨这种激酶活性结构是如何动态调节激素和生长因子和致癌突变的干扰。我们还将使用我们的单分子光遗传学方法来探测活性结构的连接性、稳健性和灵敏度。 我们预计这项研究的成功完成将产生一套新的，变革性的技术，有可能改变在细胞组织背景下研究生化过程的方式。最重要的是，建立这种新的概念框架的空间组织的活动架构应该产生一个范式转变，就我们的理解的行为的活性分子在其原生环境。表征癌症中的“失调”活性结构应导致开发针对这种失调的更有效的治疗性治疗。

 DESCRIPTION (provided by applicant): The assembly/disassembly and enzymatic activities of protein nanomachines underlie all cellular functions, and dysregulated nanomachines are the ultimate culprits in cancer. Our proposed research seeks to establish a new conceptual framework to specifically understand the cellular organization of molecular activities. We hypothesize that cellular biochemical activities are spatially organized into an "activity architecture" via the specific organization of active molecules and their regulatory partners. This activity architecture, together with the structural and mechanical architecture of the cell, encodes all the information needed to drive cellular function. We further hypothesize that perturbations to this activity architecture, even by a few dysregulated driver molecules, could lead to detrimental effects on cellular functions such as loss of control over cell growth, divisio and death. The key technological innovation of our research program is to develop and utilize a new generation of enabling technologies for visualizing and perturbing biochemical and biophysical processes in the native environment of a living cell. Enabled by these new technologies, we will start testing our general hypothesis by elucidating the spatial organization of the enzymatic activities of protein kinases, a family of enzymes that play critical roles in normal cell physiology and tumorigenesis. We will further probe how this kinase activity architecture is dynamically modulated by hormones and growth factors and perturbed by oncogenic mutations. We will also use our single- molecule optogenetic method to probe the connectivity, robustness and sensitivity of the activity architecture. We expect the successful completion of this study will yield a suite of new, transformative technologies with the potential to change the way that biochemical processes are studied in the context of cellular organization. Most importantly, establishing this new conceptual framework of a spatially organized activity architecture should produce a paradigm shift with respect to our understanding of the behaviors of active molecules in their native environment. Characterization of "dys-organized" activity architectures in cancers should lead to the development of more effective therapeutic treatments that target such dysregulation.