Novel approaches to study immune responses to post translational modifications for cancer detection

研究癌症检测翻译后修饰免疫反应的新方法

• 批准号: 9767078
• 负责人: Karen Sue Anderson
• 金额: $ 43.97万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-21 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9767078
• 关键词: Antibodies; Antibody Response; Autoantibodies; B-Lymphocytes; Biochemical Markers; Biological Markers; Biometry; Biopsy; Blinded; Bos taurus structural-GP protein; Breast; Breast Cancer Early Detection; CLIA certified; Cancer Control; Cancer Detection; Cancer Patient; Cells; Data; Detection; Development; Devices; Diagnosis; Disease; Early Detection Research Network; Early Diagnosis; Enzyme-Linked Immunosorbent Assay; Epitopes; Extracellular Domain; GALNT3 gene; Glycoproteins; Goals; Human; Human Papillomavirus; Image; Imaging technology; Immune; Immune Sera; Immune response; Immunity; In Situ; Laboratories; Length; Malignant Neoplasms; Malignant neoplasm of lung; Mammography; Measures; Medical Oncology; Membrane Proteins; Methods; Molecular Chaperones; Morbidity - disease rate; Mucins; Ovarian; Patients; Performance; Phase; Polysaccharides; Post-Translational Protein Processing; Predictive Value; Production; Protein Glycosylation; Proteins; Proteome; Proteomics; Protocols documentation; Recombinants; Reproducibility; Research Design; Ribosomes; Screening for cancer; Screening procedure; Sensitivity and Specificity; Serum; Serum Immunologic; Structure; Suggestion; Surface; Technology; Testing; Training; Translating; Tumor Antigens; Validation; base; biomarker development; biomarker discovery; biomarker identification; cancer biomarkers; chest computed tomography; clinical biomarkers; cost; design; early detection biomarkers; gene cloning; glycoprotein structure; glycoproteomics; glycosylation; glycosyltransferase; immunogenic; improved; improved outcome; malignant breast neoplasm; mortality; novel; novel strategies; overexpression; polypeptide; potential biomarker; prevent; programs; protein expression; quality assurance; rapid detection; rapid technique; screening; seal; sugar; sugar nucleotide; synthetic peptide; tool; triple-negative invasive breast carcinoma; tumor; validation studies

Project Summary/Abstract Despite advances in screening and treatment, mortalities from breast and lung cancers have remained high in the US over the last 20 years. It is widely accepted that early detection is critical to improving outcomes in both diseases. Both also rely on imaging for screening, but false positive and false negative detection are associated with unnecessary biopsies, missed diagnoses, and costs. There is an urgent need for biochemical markers that improve the performance of imaging technologies. Our laboratories have been successful at identifying useful cancer biomarkers by exploiting patients’ own ability to produce antibodies against tumor- associated antigens (TAA), referred to as tumor-associated autoantibodies (TAAb). With prior EDRN support, we developed high-throughput programmable protein display methods for the rapid detection and validation of autoantibody biomarker signatures in breast and lung cancers. Our breast cancer TAAb biomarkers have been licensed and integrated into Videssa™ Breast that is now available as CLIA-certified test. Our triple negative breast cancer markers have been validated in blinded phase 2 multicenter validation studies. These demonstrate the great utility of TAAb in cancer early detection. However, the sensitivities of most TAAbs is moderate and there is a suggestion that greater sensitivity and specificity could be obtained by examining TAAb directed at aberrantly modified proteins in cancers. Our central hypothesis is that aberrant protein glycosylation, a hallmark of breast and lung cancers, induces glycoprotein-specific TAAb that can be measured as specific serum biomarkers of these cancers. Alterations in glycosylation are highly immunogenic, and there is strong historical evidence for significant antibody responses to cancer-altered glycoproteins. However, all current protein (or polypeptide) display tools allow limited or no post-translational modification. This historical roadblock has prevented the identification of these biomarkers because of the lack of screening methods that test immunogenic structural glycoproteins. We introduce a tool for the high-throughput display of full-length proteins decorated with cancer-specific O-glycan structures. This will revolutionize the opportunity to screen glycan-protein epitopes in their natural context. Our team comprises strong expertise in functional proteomics, biomarker development, glycoproteomics, medical oncology and biostatistics. Targeted proteins will include: the extra-cellular domains of relevant single pass membrane proteins, proteins known to be O-glycosylated and overexpressed in the two cancers, and all known mucins. They will be translated in situ using human ribosomes and chaperone proteins and then systematically decorated with Tn and STn O-GalNAc-type glycans by consecutive addition of recombinant glycosyltransferases and sugar nucleotides to mirror what occurs in the two cancers. Adhering to the principles of PRoBE design, we will screen these arrays with cancer patient and control sera. Our study will focus on cancer patients and non-cancer subjects with positive imaging findings. Study design will include Phase I discovery (arrays/ELISA) and Phase II validation using ELISA.

项目概要/摘要 尽管筛查和治疗取得了进步，但乳腺癌和肺癌的死亡率仍然很高 过去 20 年来，人们普遍认为早期发现对于改善这两种疾病的结果至关重要。 两者都依赖影像学进行筛查，但存在假阳性和假阴性检测。 与不必要的活检、漏诊和费用相关的迫切需要生化。 我们的实验室在提高成像技术性能的标记物方面取得了成功。 通过利用患者自身产生抗肿瘤抗体的能力来识别有用的癌症生物标志物 相关抗原（TAA），称为肿瘤相关自身抗体（TAAb），在先前的 EDRN 支持下， 我们开发了高通量可编程蛋白质展示方法，用于快速检测和验证 我们的乳腺癌 TAAb 生物标志物已成为乳腺癌和肺癌的自身抗体生物标志物特征。 已获得许可并集成到 Videssa™ Breast 中，现已作为经过 CLIA 认证的三阴性检测提供。 乳腺癌标志物已在盲法 2 期多中心验证研究中得到验证。 证明 TAAb 在癌症早期检测中的巨大效用然而，大多数 TAAb 的敏感性较低。 中等，有人建议通过检查可以获得更高的敏感性和特异性 TAAb 针对癌症中异常修饰的蛋白质。我们的中心假设是异常蛋白质。 糖基化是乳腺癌和肺癌的标志，可诱导可测量的糖蛋白特异性 TAAb 作为这些癌症的特异性血清生物标志物，糖基化的改变具有高度免疫原性，并且存在。 是对癌症改变的糖蛋白产生显着抗体反应的强有力的历史证据。 当前的蛋白质（或多肽）显示工具允许有限的或不允许翻译后修饰。 由于缺乏筛选方法，阻碍了这些生物标志物的识别。 我们引入了一种用于全长高通量展示的工具。 用癌症特异性 O-聚糖结构修饰的蛋白质这将彻底改变筛选的机会。 我们的团队在功能蛋白质组学方面拥有丰富的专业知识， 生物标志物开发、糖蛋白组学、医学肿瘤学和生物统计学将包括： 相关单程膜蛋白的胞外结构域，已知为 O-糖基化的蛋白 并在两种癌症中过度表达，并且所有已知的粘蛋白都将使用人类进行原位翻译。 核糖体和伴侣蛋白，然后用 Tn 和 STn O-GalNAc 型聚糖系统地修饰 通过连续添加重组糖基转移酶和糖核苷酸来反映在 遵循 PRoBE 设计原则，我们将针对癌症患者和两种癌症进行筛选。 我们的研究将重点关注具有阳性影像学结果的癌症患者和非癌症受试者。 研究设计将包括第一阶段发现（阵列/ELISA）和使用 ELISA 的第二阶段验证。