Glycosylation and Immune Evasion in Urologic Tumors

泌尿系统肿瘤中的糖基化和免疫逃避

• 批准号: 10394718
• 负责人: JAMES D. BROOKS
• 金额: $ 58.89万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10394718
• 关键词: Binding; Binding Proteins; Biological; Biological Assay; Biological Markers; Biology; CRISPR interference; Cancer Detection; Cancer Patient; Cancerous; Cell surface; Cells; Clear cell renal cell carcinoma; Clinical; Collection; Data; Data Set; Development; Environment; Event; Failure; Family member; Gene Expression; Genes; Glycoproteins; Goals; Histologic; Human; Immune; Immune Evasion; Immune checkpoint inhibitor; Immune signaling; Immune system; Immunoglobulins; Immunohistochemistry; Immunosuppression; Immunotherapy; Investigation; Kidney Neoplasms; Knowledge; Lead; Lectin; Ligands; Light; Malignant Neoplasms; Malignant neoplasm of prostate; Measurement; Measures; Methods; Modification; Natural Immunity; Nature; Normal tissue morphology; Operative Surgical Procedures; Outcome; Patient Selection; Patient-Focused Outcomes; Patients; Pattern; Prognosis; Prognostic Marker; Prostate; Prostatic Neoplasms; Protein Isoforms; Proteins; Recording of previous events; Renal Cell Carcinoma; Renal carcinoma; Resected; Role; Sampling; Shapes; Sialic Acids; Sialoglycoproteins; Site; Stains; Surface; T-Lymphocyte; Testing; Therapeutic; Time; Tissue Microarray; Tissue Sample; Tissues; Tumor Tissue; Tumor-infiltrating immune cells; Urologic Cancer; Validation; adaptive immunity; base; cancer biomarkers; cancer cell; cancer type; clinical translation; companion diagnostics; comparative; diagnostic biomarker; disease prognosis; experimental study; genome wide screen; genome-wide; glycoproteomics; glycosylation; immune activation; immune function; immunogenicity; immunological synapse; immunoreactivity; inhibitor; interest; neoplastic cell; overexpression; prognostic indicator; prostate cancer cell; receptor; sialic acid binding Ig-like lectin; sialylation; success; sugar; transcriptome sequencing; tumor; tumor microenvironment; tumorigenesis; urologic

PROJECT SUMMARY/ABSTRACT The ability of tumor cells to evade the immune system is a well-known, yet poorly understood phenomenon in early cancer development. Despite promising immunotherapy strategies that have emerged from targeting these interactions, there is relatively little known about the complete repertoire of receptor-ligand interactions that contribute to immune evasion. We seek to understand how glycosylation, a well-established aberrant modification in cancer, aids cancer cells in evading the immune system. Identification of glycoproteins that modulate immune function could lead to new types of therpaies and could also serve as companion diagnostic biomarkers to guide patient selection of immunotherapies at an early time point in prostate cancer and clear cell renal cell carcinoma. First, because sialic acid is known to be overexpressed on the surface of cancer cells, we will use intact glycoproteomics methods developed in-house to enrich and identify sialoglycoproteins from cancerous and matched healthy tissues from patients. Quantitative comparative analyses will reveal changes in sialoglycoprotein expression and illuminate candidate ligands for sialic acid-binding proteins in the tumor microenvironment that potentially contribute to immune inactivation. Correlation of these glycoproteomic datasets with RNA-seq data focused on glycogene expression will bolster the assignment of specific glycoforms as cancer biomarkers. Second, using immunohistochemistry and CODEX methods, we will analyze expression levels of sialic acid-binding immunoglobulin-type lectin (Siglec) receptor proteins on tumor- resident immune cells and cross-correlate the findings with RNA-seq data as well as immune cell markers. We will also probe for the presence of ligands for various Siglec isoforms on tumor cell surfaces and obtain spatial information about their distribution on immune cells in intact tumor tissue. For any Siglecs identified as prominently displayed on immune cells in the tumor environment, we will develop cell-based assays to probe their contribution to tumor cell immunoreactivity. Third, we will perform a genome-wide screening using CRISPRi to identify genes that facilitate the binding of Siglecs to cancer cells. Finally, we will correlate the datasets from Aims 1, 2, and 3 with patient outcomes in a larger set of tissue samples contained on a tissue microarray, and evaluate their utility as prognostic indicators.

项目摘要/摘要 肿瘤细胞逃避免疫系统的能力是众所周知但知之甚少 早期癌症发展中的现象。尽管有希望的免疫疗法策略已经出现 从针对这些相互作用来看，关于受体配体的完整曲目鲜为人知 有助于免疫逃避的相互作用。我们试图了解糖基化是如何建立的 癌症的异常修饰，有助于癌细胞逃避免疫系统。识别 调节免疫功能的糖蛋白可能会导致新型的缺点，也可以作为 伴侣诊断生物标志物在早期指导患者选择免疫疗法的时间 前列腺癌和透明细胞肾细胞癌。 首先，由于已知唾液酸在癌细胞的表面过表达，因此我们将使用完整 内部开发的糖蛋白质组学方法是为了富集和鉴定从癌和 匹配患者的健康组织。定量比较分析将揭示 唾液酸糖蛋白表达和照亮肿瘤中唾液酸结合蛋白的候选配体 可能导致免疫失活的微环境。这些糖蛋白酶的相关性 带有RNA-seq数据的数据集以糖基分析表达表达，将加强特定的分配 糖型作为癌症生物标志物。第二，使用免疫组织化学和法典方法，我们将 分析唾液酸结合免疫球蛋白型凝集素（SIGLEC）受体蛋白上的表达水平 驻留的免疫细胞并与RNA-seq数据以及免疫细胞标记物互相交叉。我们 还将探测是否存在针对肿瘤细胞表面上各种Siglec同工型的配体的存在并获得空间 有关它们在完整肿瘤组织中免疫细胞上分布的信息。对于任何被确定为 我们将在肿瘤环境中突出显示在免疫细胞上，我们将开发基于细胞的测定以探测 它们对肿瘤细胞免疫反应性的贡献。第三，我们将使用 CRISPRI鉴定促进Siglecs与癌细胞结合的基因。最后，我们将关联 来自AIM 1、2和3的数据集，在较大的组织样品中具有患者的结果 微阵列，并评估它们作为预后指标的效用。