Project 2: Cellular topography and function of the breast cancer tissue microenvironment

项目2：乳腺癌组织微环境的细胞形态和功能

• 批准号: 10704687
• 负责人: Robert michael Angelo
• 金额: $ 34.15万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-14 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10704687
• 关键词: 2-Oxoglutarate 5-Dioxygenase Procollagen-Lysine; Antibodies; Automobile Driving; Avidity; Binding; Biopsy; Breast cancer metastasis; CRISPR interference; Cell Communication; Cells; Clinical; Collagen; Data; Data Set; Deaminase; Disease Progression; Distant; Enzymes; Estrogen receptor positive; Excision; Extracellular Matrix; Family; Fibroblasts; Frequencies; Hydroxylation; Image; Immune; Immune Evasion; Immunosuppression; Invaded; Label; Libraries; Location; Machine Learning; Macrophage; Maps; Mass Spectrum Analysis; Metabolic; Metals; Metastatic breast cancer; Methods; Mixed Function Oxygenases; Modeling; Modification; Molecular; Morphology; Multiplexed Ion Beam Imaging; Neighborhoods; Neoplasm Metastasis; Nivolumab; Normal tissue morphology; Operative Surgical Procedures; Organoids; Outcome; Pathogenesis; Patients; Peptide Hydrolases; Peptides; Play; Polysaccharides; Population; Process; Procollagen-Proline Dioxygenase; Protein Glycosylation; Proteins; Proteomics; Resolution; Resources; Risk; Role; Sampling; Site; Specimen; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Stains; Stromal Cells; Structure; Testing; Therapeutic; Time; Tissue Sample; Tissue imaging; Tissues; Tumor Cell Migration; Visualization; Work; biobank; breast cancer progression; breast imaging; cell type; clinically significant; collagenase; crosslink; glycosylation; glycosyltransferase; high risk; imaging modality; immune activation; malignant breast neoplasm; mass spectrometric imaging; neoplastic cell; permissiveness; prevent; receptor; recruit; response; sialylation; tool; transcriptomics; triple-negative invasive breast carcinoma; tumor; tumor microenvironment

Abstract/Project Summary Breast cancer (BC) metastasis is an emergent feature that occurs when the tumor’s ability to recruit metabolic resources, avoid immune activation, and disseminate to distant sites exceeds the capacity of surrounding normal tissue to prevent these processes. In line with this, work over the last decade has demonstrated the crucial role played by the tumor microenvironment (TME) in promoting or deterring BC progression. Tumor cell migration and immune recruitment have both been shown to be heavily influenced by fibroblasts and the surrounding extracellular matrix (ECM) they produce. Additionally, protein glycosylation has been shown to modulate these interactions. To understand how tumor glycosylation and ECM remodeling interact to potentiate BC metastasis, we will use spatial transcriptomics and two complementary mass spectrometry methods, MIBI-TOF and MALDI, to identify glycan-dependent, cell-cell, and cell-ECM interactions that shift the TME toward tumor permissive states. All three analyses will be performed on spatially-coregistered serial sections from the same tissue blocks. In doing so, comprehensive single-cell maps of each tissue sample constructed by MIBI-TOF can be directly superimposed with de novo proteomic and transcriptomic data. We will map and enumerate the lineage and major functional subsets of tumor and stromal cells with respect to relevant therapeutic and molecular parameters to understand how the BC TME evolves with disease progression. These features will be overlaid with de novo imaging of tissue glycans to identify potential mechanisms of immune evasion that involve tumor sialoglycans and macrophage-bound SIGLECs. The frequency and spatial enrichment of these features will be correlated with spatial transcriptomics data to identify regulatory glycosyltransferases promoting these interactions. Next, ECM-MALDI and MIBI-TOF data will be used to identify how collagen type, hydroxylation, and crosslinking shift in coordination with the collagen structure and function of neighboring cell populations. In particular, we will focus on understanding how the activity of two families of enzymes, prolyl and lysyl hydroxylases, drive structural changes in the ECM that promote BC metastasis. The clinical significance of these extracted cellular and molecular definitions of ECM remodeling will be assessed with respect to metastatic risk, stage, and IC subtype. Taken together, this work will provide an unprecedented view into how TME structure and cell-cell interactions between tumor and stroma relate to specific facets of the tumor ECM and glycome.

摘要/项目摘要 乳腺癌（BC）转移是一种紧急特征，当肿瘤募集代谢能力时会发生 资源，避免免疫激活并传播以剖析地点超过周围正常的能力 组织以防止这些过程。与此一致，在过去的十年中的工作已经证明了至关重要的角色 肿瘤微环境（TME）在促进或确定BC进展方面发挥了作用。肿瘤细胞迁移 并且免疫招募都被证明受成纤维细胞和周围的影响很大 它们产生的细胞外基质（ECM）。此外，已证明蛋白质糖基化可以调节这些 互动。要了解肿瘤糖基化和ECM重塑如何相互作用，以使BC转移相互作用， 我们将使用空间转录组学和两种完整的质谱法，Mibi-Tof和Maldi， 确定依赖聚糖的，细胞细胞和细胞ECM相互作用，将TME转移到肿瘤允许的 国家。所有三个分析都将对来自相同组织块的空间孔仪连续切片进行。 这样，通过MIBI-TOF构建的每个组织样品的全面单细胞图可以直接 与从头蛋白质组学和转录组数据叠加。我们将绘制并列举谱系和 相对治疗和分子相对于肿瘤和基质细胞的主要功能子集 参数以了解随着疾病进展的BC TME演变。这些功能将被覆盖 通过对组织聚糖的从头成像，以识别涉及肿瘤的免疫驱虫的潜在机制 唾液聚糖和巨噬细胞结合的siglecs。这些功能的频率和空间增强将是 与空间转录组数据相关，以识别促进这些调节性糖基转移酶 互动。接下来，将使用ECM-Maldi和MiBI-TOF数据来确定胶原蛋白类型，羟基化和如何如何 与相邻细胞群体的胶原蛋白结构和功能的协调转移。在 特别是，我们将专注于了解两个酶，丙酰和赖氨酸的活性如何 羟化酶驱动ECM中促进BC转移的结构变化。这些的临床意义 ECM重塑的细胞和分子定义将根据转移风险进行评估， 阶段和IC亚型。综上所述，这项工作将为TME结构提供前所未有的观点 肿瘤与基质之间的细胞细胞相互作用与肿瘤ECM和Glyce的特定方面有关。