Identifying antigens bound by novel scFvs targeting all subtypes of mesothelioma

识别针对间皮瘤所有亚型的新型 scFv 结合的抗原

• 批准号: 8209276
• 负责人: BIN LIU
• 金额: $ 31.1万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8209276
• 关键词: Address; Affinity; Antibodies; Antibody Affinity; Antigen Targeting; Antigens; Asbestos; Bacteriophages; Behavior; Binding; Biodistribution; Biological Assay; Blood Vessels; Cell membrane; Cell surface; Cells; Clinic; Clinical Trials Design; Collaborations; Complementary DNA; Detection; Development; Diagnosis; Diagnostic; Disease; Drug Kinetics; Engineering; Enzyme-Linked Immunosorbent Assay; Epitopes; Eukaryotic Cell; Exhibits; Exposure to; Funding; Future; Goals; Human; In Situ; In Vitro; Lead; Libraries; Life; Ligands; Light; MCAM gene; Malignant - descriptor; Malignant Fibrous Mesothelioma; Malignant mesothelioma; Maps; Mass Spectrum Analysis; Mediating; Mesothelioma; Mesothelium; Methods; Molecular; National Cancer Institute; National Institute for Occupational Safety and Health; Occupations; Patients; Positioning Attribute; Post-Translational Protein Processing; Probability; Property; Proteins; Proteome; Radioimmunotherapy; Research Personnel; Route; Screening procedure; Specificity; Staging; Surface; Surface Antigens; Symptoms; Therapeutic; Tissues; Tumor Antigens; Work; X-Ray Computed Tomography; Xenograft Model; Yeasts; abstracting; antigen binding; assay development; base; combinatorial; disorder subtype; expression cloning; human monoclonal antibodies; improved; in vivo Model; member; mesothelin; neoplastic cell; novel; novel diagnostics; novel therapeutics; outcome forecast; prognostic; single photon emission computed tomography; small molecule; sound; therapeutic development; therapeutic target; tumor

Project Summary/Abstract: The objective of this proposal is to establish the molecular identities of a panel of internalizing mesothelioma cell surface antigens that are bound by a panel of novel internalizing human single chain antibodies (scFvs) that target both epithelioid and sarcomatoid (a particularly recalcitrant form) mesothelioma. The proposed study is built upon our recent work where we have selected a combinatorial human antibody library on live mesothelioma cells and identified a panel of internalizing scFvs that bind to all subtypes of mesothelioma cells in situ with no binding to normal mesothelium, and mediate efficient intracellular delivery of small molecule payloads to both epithelioid and sarcomatoid mesothelioma cells in vitro. We hypothesize that these scFvs define novel mesothelioma antigens that have significantly greater specificity, subtype coverage and therapeutic potential than currently known antigens. Identification of these novel mesothelioma antigens would (1) allow further engineering of the lead antibodies to improve affinity, specificity, pharmacokinetics and biodistribution, (2) allow development of additional antibodies targeting non-overlapping epitopes of the same antigen, which is often required for diagnostic assay development, (3) enhance our understanding of tumor behaviors that involve the cell membrane, which may lead to identification of additional novel targets for further therapeutic development, and (4) provide a sound rationale for clinical trial design such as pre-screening of patients based on target expression. We are uniquely positioned to accomplish this goal as we have developed and adapted effective methods to identify tumor antigens targeted by scFvs. Key to this proposal is our newly developed novel antigen identification strategies based on eukaryotic cell surface display of the human proteome. In addition, we have established functional collaborations with Drs. Burlingame and Chalkley and their National Mass Spectrometry Facility at UCSF to identify tumor antigens that are difficult to identify by expression cloning methods such as post-translationally modified antigens. We propose to use the methods and strategies that we have developed and adapted to systematically identify mesothelioma cell surface antigens targeted by our panel of internalizing human scFvs.

项目摘要/摘要： 该提案的目的是确定一组内化间皮瘤的分子特性 与一组新型内化人单链抗体 (scFv) 结合的细胞表面抗原 针对上皮样间皮瘤和肉瘤样间皮瘤（一种特别顽固的形式）。 拟议的研究是建立在我们最近的工作基础上的，我们选择了一个组合人类 活间皮瘤细胞上的抗体库，并鉴定了一组与所有亚型结合的内化 scFv 原位间皮瘤细胞不与正常间皮结合，并介导有效的细胞内递送 体外上皮样和肉瘤样间皮瘤细胞的小分子有效负载。我们假设 这些 scFv 定义了具有显着更高特异性、亚型的新型间皮瘤抗原 覆盖范围和治疗潜力均高于目前已知的抗原。这些新型间皮瘤的鉴定 抗原将（1）允许对先导抗体进行进一步的工程改造，以提高亲和力、特异性、 药代动力学和生物分布，(2) 允许开发针对非重叠的其他抗体 相同抗原的表位，这通常是诊断测定开发所必需的，（3）增强我们的 了解涉及细胞膜的肿瘤行为，这可能导致识别额外的 进一步治疗开发的新靶点，以及（4）为临床试验设计提供合理的理由，例如 根据目标表达对患者进行预筛选。 我们拥有独特的优势来实现这一目标，因为我们已经开发并采用了有效的 鉴定 scFv 靶向的肿瘤抗原的方法。这个提案的关键是我们新开发的小说 基于人类蛋白质组的真核细胞表面展示的抗原识别策略。此外， 我们与 Drs 建立了功能性合作。伯林格姆和查克利以及他们的全国弥撒 加州大学旧金山分校的光谱测定设施可鉴定难以通过表达克隆鉴定的肿瘤抗原 方法例如翻译后修饰抗原。我们建议使用我们所采用的方法和策略 已开发并适应系统地识别我们的靶向间皮瘤细胞表面抗原 内化人类 scFv 小组。