Characterizing glioma heterogeneity with novel multiplexed nanoscale imaging technologies

摘要： 胶质瘤，特别是胶质母细胞瘤（GBM），是人类最致命的肿瘤之一， 恶性肿瘤，由于其侵入性，目前的常规治疗缺乏成功， 性质和异质性。我们将利用膨胀显微镜，一种新的超分辨率 显微镜方法，作为一个平台，开发新的工具，高度多重原位分析， 生成GBM空间异质性和结构的综合地图。我们将会用这些 有助于了解GBM侵袭的分子和形态表型的工具， 病理学，我们希望这将指导未来的治疗干预。 拟议的研究将包括三个目标，旨在开发新的原位 GBM组织中生物学问题驱动的分析工具：（1）全面绘制细胞图 GBM肿瘤组织内的类型和状态具有高空间分辨率。为了实现这一点，我们将 开发了一种创新的方法，用于扩增组织中RNA的高度多重读出， 基于单细胞RNA测序特征的定制原位基因表达面板。（二） 研究肿瘤细胞与周围微环境之间的纳米级相互作用 与GBM入侵有关为了实现这一点，我们将开发一种方法， 使用DNA条形码抗体库的纳米级分辨率的蛋白质成像。(3)开发一个 具有自动化和分析软件的可扩展分析平台，可整合多重RNA 和蛋白质成像。我们将把这个平台应用于病人源性肿瘤 样本，以了解入侵GBM肿瘤细胞的分子和形态表型， 以及建立GBM空间异质性的地图。这些新技术在空间上 绘制复杂组织中的分子信息不仅对了解神经胶质瘤有价值， 和癌症，它们将广泛适用于许多其他空间复杂的生物过程。

Abstract: Gliomas, with glioblastomas (GBM) in particular, are one of the most lethal human malignancies due to lack of success with current conventional treatments due to their invasive nature and heterogeneity. We will leverage expansion microscopy, a novel super-resolution microscopy method, as a platform to develop novel tools for highly multiplexed in situ analysis to generate comprehensive maps of GBM spatial heterogeneity and structure. We will use these tools to help understand the molecular and morphological phenotypes of GBM invasion and pathology, which we hope will guide future therapeutic interventions. The proposed research will consist of three aims which seeks to develop novel in situ analysis tools driven by biological questions in GBM organization: (1) Comprehensively map cell types and states within GBM tumor tissue with high spatial resolution. To enable this, we will develop an innovative method for highly multiplexed readout of RNA in expanded tissues with tailored in situ gene expression panels based on single-cell RNA sequencing signatures. (2) Study the nanoscale interactions between tumor cells and their surrounding microenvironment that are implicated GBM invasion. To accomplish this, we will develop an approach for multiplexed protein imaging with nanoscale resolution using DNA barcoded antibody libraries. (3) Develop a scalable analysis platform with automation and analysis software to integrate multiplexed RNA and protein imaging in the same sample. We will apply this platform to patient derived tumor samples to understand the molecular and morphological phenotypes of invading GBM tumor cells, as well as build a map of the spatial heterogeneity of GBM. These novel technologies to spatially map molecular information in complex tissues will not only be invaluable for understanding glioma and cancer, they will be broadly applicable to many other spatially complex biological processes.