Omic and Multidimensional Spatial Atlas of Metastatic Breast Cancer

项目摘要 我们将继续开发我们的结构和多维空间(OMS)地图集，使我们能够发现 转移性乳腺癌患者个体化耐药机制的研究 当前一代的靶向治疗组合和免疫检查点抑制剂。OMS地图集是 动机是这些治疗通常只在转移性环境中暂时有效。 可能的耐药机制可能是肿瘤细胞固有的或来自不同的微环境。 肿瘤细胞生活的地方。OMS图谱侧重于在两个电流中阐明这些阻力机制 产生的临床情景：(A)激素受体阳性的乳腺癌(HRBC)正在接受 CDK4/6抑制剂联合内分泌治疗，以及(B)三阴性乳腺癌(TNBC) 使用PARP抑制剂和免疫调节剂治疗。我们正在通过以下方式实现这一目标： 四个协调一致的单位。一个生物医学单位正在前瞻性地收集和分发纵向临床数据 来自两个转移性乳腺癌队列的30例患者的信息、血液和活检。一个 鉴定单位正在分析(A)OCT冷冻标本，使用单细胞DNA-SEQ和单细胞ATAC- SEQ以阐明单细胞中空间定义的基因组变化和染色质可及性，(B)福尔马林固定， 石蜡包埋(FFPE)标本多重免疫组织化学(MIHC)检测免疫功能 微环境和循环免疫荧光法(CycIF)评价人血清白蛋白的组成和分子状态 肿瘤细胞及其微环境，以及(C)多聚甲醛固定，树脂包埋(PFRE)样品 用聚焦离子束扫描电子显微镜(FIB-SEM)观察2D的超微结构变化 图像和目标3D图像。同一肿瘤样本的基因组特征由SMMART提供 程序。数据分析股正在开发和部署工具，以(A)管理、分析和可视化组学 和成像数据集，(B)通过横向映射将组学和成像数据集集成在一起，以创建单个 肿瘤时间点地图和量化肿瘤细胞亚群的系统生物学功能，以及(C)探索 治疗前后肿瘤图谱的差异以揭示耐药机制。这个 行政股协助协调、运作、互动和评估OMS内的活动 阿特拉斯以及OMS阿特拉斯科学家和Htan之间的合作。在这个项目中，我们将完成我们计划的OMS地图集 项目通过以下目标：(1)使用我们的组学对另外五个患者病例进行表征 和多尺度成像分析；(2)将我们剩余的数据集和元数据上传到Htan数据协调 中心；以及(3)在我们的atlas案例数据集以及Htan跨网络中执行综合数据分析 项目数据集，以确定治疗中的肿瘤变化以及这些变化对乳腺癌的影响 治疗。通过实现这些目标，我们将完成并充分共享我们的30个多模式空间地图集 正在接受治疗的转移性乳腺癌病例。

PROJECT ABSTRACT We will continue development of our Omic and Multidimensional Spatial (OMS) Atlas that enables discovery of mechanisms of resistance that arise in individual patients with metastatic breast cancer during treatment with current generation targeted therapeutic combinations and immune checkpoint inhibitors. The OMS Atlas is motivated by the fact that these treatments typically are only transiently effective in the metastatic setting. Possible resistance mechanisms may be intrinsic to the tumor cells or derive from the diverse microenvironments in which the tumor cells live. The OMS Atlas focuses on elucidating these resistance mechanisms in two current generation clinical scenarios: (a) hormone-receptor positive breast cancer (HRBC) undergoing treatment with a CDK4/6 inhibitor in combination with endocrine therapy, and (b) triple negative breast cancer (TNBC) undergoing treatment with a PARP inhibitor and an immunomodulatory agent. We are accomplishing this through work via four coordinated units. A Biospecimen Unit is prospectively collecting and distributing longitudinal clinical information, blood, and biopsies from 30 cases across the two metastatic breast cancer cohorts. A Characterization Unit is analyzing (a) OCT frozen specimens using single-cell DNA-seq and single-cell ATAC- seq to elucidate spatially defined genomic changes and chromatin accessibility in single cells, (b) formalin fixed, paraffin embedded (FFPE) specimens using multiplex immunohistochemistry (mIHC) to assess the immune microenvironment and cyclic Immunofluorescence (cycIF) to assess the composition and molecular states of tumor cells and their microenvironments, and (c) paraformaldehyde fixed, resin embedded (PFRE) specimens using a Focused Ion Beam Scanning Electron Microscope (FIB-SEM) to identify ultrastructural changes in 2D images and targeted 3D images. Omic characterization of the same tumor samples is provided by the SMMART Program. A Data Analysis Unit is developing and deploying tools to (a) manage, analyze, and visualize omics and imaging datasets, (b) integrate omics and imaging datasets through crosswise mapping to create single timepoint tumor maps and quantify systems biological functions of tumor cellular subpopulations, and (c) explore differences between pre- and on/post-treatment tumor maps to reveal mechanisms of resistance. The Administrative Unit facilitates the coordination, operation, interaction, and evaluation of activities within the OMS Atlas and between OMS Atlas scientists and the HTAN. In this project, we will complete our planned OMS Atlas project through the following Aims: (1) perform characterization of five additional patient cases using our omics and multiscale imaging assays; (2) upload our remaining datasets and metadata to the HTAN data coordinating center; and (3) perform integrated data analysis across our atlas case datasets as well as HTAN trans-network project datasets to identify tumor changes on therapy and the implications of those changes on breast cancer treatment. By accomplishing these aims we will complete and fully share our multimodal spatial atlas of 30 metastatic breast cancer cases undergoing treatment.