Technology Development Unit

技术开发部

• 批准号: 10375270
• 负责人: Hao F Zhang
• 金额: $ 85.62万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-10 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10375270
• 关键词: 3-Dimensional; Address; Beds; Behavior; Budgets; Caliber; Cancer Biology; Cell Nucleus; Cell physiology; Cells; Chemoresistance; Chromatin; Chromatin Modeling; Chromatin Structure; Clustered Regularly Interspaced Short Palindromic Repeats; Color; Communities; Computer Models; Crowding; DNA; Data; Development; Electron Microscopy; Ensure; Epigenetic Process; Etiology; Family; Feedback; Gene Expression; Genes; Genetic Transcription; Genome Mappings; Genomics; Goals; Heterogeneity; Image; Label; Link; Location; Longterm Follow-up; Malignant Neoplasms; Messenger RNA; Methods; Microscopy; Modality; Modeling; Modification; Molecular; Molecular Analysis; Molecular Computations; Molecular Conformation; Molecular Target; Nucleosomes; Optics; Pattern; Phenotype; Photons; Physics; Polymers; Population; Preparation; Process; Quantum Dots; Reaction; Regulator Genes; Research; Research Personnel; Resistance; Resolution; Sampling; Scanning; Structure; Techniques; Technology; Testing; Therapeutic; Time; Tissues; Transmission Electron Microscopy; Universities; Work; anticancer research; base; cancer cell; cancer stem cell; cellular imaging; chemotherapy; chromatin remodeling; genome sequencing; genomic locus; histone modification; imaging platform; light intensity; microscopic imaging; molecular dynamics; molecular imaging; molecular modeling; molecular scale; nano; nanoassembly; nanoimaging; nanoscale; nanosensors; new technology; next generation; novel; predictive modeling; response; role model; single molecule; spatiotemporal; spectroscopic imaging; statistics; stem; success; targeted imaging; technology development; theories; three dimensional structure; transcriptional reprogramming

Technology Development Unit: PROJECT SUMMARY The overall goal of the U54 Northwestern University Center for Chromatin Nanoimaging in Cancer (NU-CCNIC) is to develop and deploy a multi-scale chromatin nanoimaging platform together with molecular analyses and computational modeling to characterize chromatin structure and transcriptional patterns associated with cancer stem cells (CSCs) and the chemoresistance phenotype. We seek to fill three technology gaps in developing such a nanoimaging platform. (1) The multi-scale challenge stems from the need to investigate how chromatin regulates gene expression across a wide range of scales, from the diameter of DNA (~2 nm) to the size of nucleosomes (~10 nm) to chromatin domains (~100 nm) to cell nuclei (~10 µm) and to a population of cells to account for intercellular heterogeneity. (2) The multiplexed molecular imaging challenge arises due to a large number of critical genes and molecular regulators that need to be co-registered with the 3D chromatin structural data. And (3) The temporal dynamics challenge requires the ability to work across a wide range of time scales to study short-term processes that may occur within minutes, such as chromatin remodeling, and long-term processes that may progress over weeks, such as the emergence of chemoresistance in cancer cells. No single technique satisfies all three requirements. Our solution to address these three challenges is to organically correlate three families of technologies: chromatin scanning transmission electron microscopy (ChromSTEM), spectroscopic single-molecule localization microscopy (sSMLM), and partial wave spectroscopic (PWS) nanosensing microscopy. After registering these technologies with 1 nm precision, they will collectively satisfy the multi-scale, multiplexing, and dynamics requirements. We will achieve three objectives in TECH. (1) Develop next-generation sSMLM that enables, in principle, unlimited multiplexing with the highest photon utilization and at 1 nm spatial precision. (2) Develop correlative ChromSTEM-sSMLM-PWS imaging that enables cross-modality registration at 1 nm precision assisted by novel quantum dot-based nano assembly fiduciary markers. And (3) Develop a multi-scale molecular modeling platform to elucidate the etiological relationship between chromatin structure and transcriptional reprogramming in CSCs imaged with the nanoimaging platform. The proposed technology development is driven by the needs of the Research Test-Bed unit (RTB) to elucidate epigenetic and chromatin drivers of transcriptional plasticity in CSC processes, in particular the development of adaptive resistance to chemotherapy, and to explore the feasibility of reprogramming CSCs out of the stem-states as a therapeutic strategy. Continuous feedback from RTB investigators will be integrated with the TECH platform's optimization, enabling, in the longer-term, our U54 technologies to impact the broader cancer research community within and beyond the Cellular Cancer Biology Imaging Research Network.

技术开发单位：项目概要 U54西北大学癌症染色质纳米成像中心（NU-CCNIC）的总体目标 旨在开发和部署多尺度染色质纳米成像平台以及分子分析和 用于表征与癌症相关的染色质结构和转录模式的计算模型 干细胞（CSC）和化疗耐药表型。我们寻求填补开发中的三个技术空白 这样的纳米成像平台。 (1) 多尺度挑战源于需要研究染色质如何 在广泛的尺度上调节基因表达，从 DNA 的直径 (~2 nm) 到 DNA 的大小 核小体 (~10 nm) 到染色质结构域 (~100 nm) 到细胞核 (~10 µm) 以及细胞群 解释细胞间的异质性。 (2) 多重分子成像的挑战是由于大量的 需要与 3D 染色质结构共同注册的关键基因和分子调节因子的数量 数据。 (3) 时间动态挑战需要能够在广泛的时间尺度上工作 研究可能在几分钟内发生的短期过程，例如染色质重塑，以及长期过程 可能会持续数周的过程，例如癌细胞中出现化疗耐药性。无单 技术满足所有三个要求。 我们应对这三个挑战的解决方案是将三个技术系列有机地关联起来： 染色质扫描透射电子显微镜 (ChromSTEM)、光谱单分子定位 显微镜（sSMLM）和部分波谱（PWS）纳米传感显微镜。注册完这些之后 1 nm精度的技术，将共同满足多尺度、多路复用和动态 要求。我们将实现科技领域的三个目标。 (1) 开发下一代 sSMLM，使 原理上，无限复用具有最高的光子利用率和 1 nm 的空间精度。 (2) 开发 相关 ChromSTEM-sSMLM-PWS 成像，可实现 1 nm 精度的跨模态配准 在新型基于量子点的纳米组装信托标记的协助下。 (3) 开发多尺度 分子建模平台阐明染色质结构与染色质之间的病因关系 使用纳米成像平台成像的 CSC 中的转录重编程。提议的技术 开发是由研究试验台单元 (RTB) 的需求驱动的，以阐明表观遗传和染色质 CSC 过程中转录可塑性的驱动因素，特别是适应性抗性的发展 化疗，并探索将 CSC 重编程出干状态作为治疗方法的可行性 战略。 RTB调查员的持续反馈将与TECH平台的优化相结合， 从长远来看，我们的 U54 技术能够影响更广泛的癌症研究界 超越细胞癌症生物学成像研究网络。