Northwestern University Center for Chromatin NanoImaging in Cancer (NU-CCNIC)

总体：项目总结 癌症干细胞（CSC）在促进肿瘤对治疗的抵抗和治疗后的复发中起着关键作用。 治疗这对开发成功的抗癌疗法提出了关键障碍。 转录重编程和可塑性在CSC状态内外起着关键作用，这反过来又是 依赖于染色质的三维（3D）结构的调节功能，表观遗传状态， 和其他分子事件。我们对CSC基本生物学的理解一直受到需要的阻碍， 对于细胞纳米级成像技术，提供关于3D的高度详细的结构信息， 染色质组织和许多分子调节因子和事件的高度多重分子成像 参与CSC进程。我们建议建立西北大学染色质中心 癌症纳米成像（NU-CCNIC），以解决细胞纳米成像的这一基本技术差距 并部署新技术，以解决CSC生物学的基本知识差距。中心 汇聚了细胞纳米成像，计算成像，分子建模，计算 基因组学、CSC生物学和肿瘤学。该中心将开发、测试、验证、部署一个集成的 和共同注册的多尺度染色质纳米成像平台，将包括三个“嵌套娃娃”成像 技术：染色质扫描透射电子显微镜，光学光谱超分辨率 nanoscopy纳米，and optical光学spectroscopy光谱nanosensing纳米sensing传感. Nanoimaging Platform将实现定量成像 染色质结构和高度多重的分子和基因特异性定位，在最基本的 长度尺度接近1 nm的分辨率，包括统计学上显著的细胞群的成像， 在延长的时间随访时间内具有高时间分辨率的活细胞。纳米成像平台将 与计算基因组学、表观基因组学、基因组作图和预测性转录建模建立桥梁 数据集。这些技术将用于回答CSC生物学中几个长期存在的开放问题。 我们将阐明CSCs是否可以通过转录重编程起源于非CSCs，测试CSCs的作用。 染色质结构在CSC过程中促进转录可塑性的作用，并探讨 转录重编程CSC以退出干细胞状态作为一种新的治疗策略。的所有方面 通过一系列的研究，将以CSC生物试验台的需要为指导，进行技术开发 反馈回路从长远来看，这种单细胞纳米成像技术将有助于全面 了解结构，物理化学和分子基因组事件之间的复杂相互作用。 我们预计，这些融合研究将为CSC生物学提供新的见解，这是不可能的 使用任何一种方法来揭示，并为确定治疗策略开辟新的机会。

Overall: PROJECT SUMMARY Cancer stem cells (CSCs) play a critical role in fostering tumor resistance to therapies and relapse after treatment. This presents a crucial barrier to the development of successful anti-cancer therapeutics. Transcriptional reprogramming and plasticity play a critical role in and out of the CSC state, which in turn are interdependent on the regulatory function of the three-dimensional (3D) structure of chromatin, epigenetic states, and other molecular events. Our understanding of fundamental CSC biology has been hampered by the need for cellular nanoscale imaging technologies that provide both highly detailed structural information regarding 3D chromatin organization and highly multiplexed molecular imaging of the many molecular regulators and events involved in CSC processes. We propose to establish the Northwestern University Center for Chromatin Nanoimaging in Cancer (NU-CCNIC) to address this fundamental technology gap in cellular nanoscale imaging and deploy the new technologies to address the fundamental knowledge gap in CSC biology. The Center converges experts in cellular nanoscale imaging, computational imaging, molecular modeling, computational genomics, CSC biology, and oncology. The Center will develop, test, validate, iterate, and deploy an integrated and co-registered Multi-scale Chromatin Nanoimaging Platform that will comprise three “nested-doll” imaging techniques: chromatin scanning transmission electron microscopy, optical spectroscopic super-resolution nanoscopy, and optical spectroscopic nanosensing. The Nanoimaging Platform will enable quantitative imaging of chromatin structure and highly multiplexed molecular and gene-specific localization, at the most fundamental length-scale approaching 1 nm resolution, including the imaging of statistically significant cell populations and live cells with high temporal resolution over prolonged temporal follow-up times. The Nanoimaging Platform will be bridged to computational genomics, epigenomics, genome mapping, and predictive transcriptional modeling datasets. These technologies will be deployed to answer several long-standing open questions in CSC biology. We will elucidate whether CSCs can originate from non-CSCs via transcriptional reprogramming, test the role of chromatin structure in fostering transcriptional plasticity in CSC processes, and explore the possibility of transcriptionally reprogramming CSCs to exit the stem-state as a new therapeutic strategy. All aspects of the technology development will be guided by the needs of the CSC biology testbed through a series of research feedback loops. In the long term, such single-cell nanoimaging technologies will help comprehensive understanding of the complex interplay between structural, physico-chemical, and molecular genomic events. We anticipate that these convergence studies will provide new insights into CSC biology, which are impossible to reveal with the use of any single method, and open new opportunities for identifying therapeutic strategies.