Glycosylation and Immune Evasion in Urologic Tumors

泌尿系统肿瘤中的糖基化和免疫逃避

基本信息

批准号：
10152526
负责人：
JAMES D. BROOKS
金额：
$ 58.23万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
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/10152526
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 Disease 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 experimental study genome wide screen genome-wide glycoproteomics glycosylation immune activation immune function immunogenicity immunological synapse immunoreactivity inhibitor/antagonist interest neoplastic cell overexpression prostate cancer cell receptor sialic acid binding Ig-like lectin sialylation success sugar transcriptome sequencing tumor tumor microenvironment tumorigenesis urologic

项目摘要

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.

项目概要/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的早期癌症发展中的现象。尽管已经出现了有希望的免疫治疗策略针对这些相互作用，人们对受体-配体的完整功能知之甚少有助于免疫逃避的相互作用。我们试图了解糖基化这一成熟的癌症的异常修饰有助于癌细胞逃避免疫系统。鉴定调节免疫功能的糖蛋白可能会带来新型疗法，也可以作为伴随诊断生物标志物，指导患者在早期时间点选择免疫疗法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表面过度表达，因此我们将使用完整的唾液酸内部开发的糖蛋白组学方法可富集和鉴定癌症和癌症中的唾液酸糖蛋白匹配来自患者的健康组织。定量比较分析将揭示变化唾液酸糖蛋白表达并阐明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。这些糖蛋白组的相关性包含专注于糖原表达的 RNA-seq 数据的数据集将支持特定的分配糖型作为癌症生物标志物。其次，使用免疫组织化学和 CODEX 方法，我们将分析肿瘤上唾液酸结合免疫球蛋白型凝集素 (Siglec) 受体蛋白的表达水平常驻免疫细胞，并将研究结果与 RNA-seq 数据以及免疫细胞标记物交叉关联。我们还将探测肿瘤细胞表面上各种 Siglec 同种型的配体的存在，并获得空间有关它们在完整肿瘤组织中免疫细胞上的分布的信息。对于任何标识为显着地显示在肿瘤环境中的免疫细胞上，我们将开发基于细胞的测定法来探测它们对肿瘤细胞免疫反应性的贡献。第三，我们将使用以下方法进行全基因组筛选： CRISPRi 识别促进 Siglecs 与癌细胞结合的基因。最后，我们将关联来自目标 1、2 和 3 的数据集，其中包含组织中包含的更大组组织样本中的患者结果微阵列，并评估其作为预后指标的效用。