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: 7888421
• 负责人: BIN LIU
• 金额: $ 32.06万
• 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/7888421
• 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; antigen binding; assay development; base; combinatorial; disorder subtype; expression cloning; human monoclonal antibodies; improved; in vivo Model; member; mesothelin; neoplastic cell; novel; novel diagnostics; outcome forecast; prognostic; public health relevance; single photon emission computed tomography; small molecule; sound; therapeutic development; therapeutic target; tumor

DESCRIPTION (provided by applicant): 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. PUBLIC HEALTH RELEVANCE: The objective of this proposal is to establish the molecular identities of a panel of mesothelioma cell surface antigens that are bound by a panel of novel internalizing human single chain antibodies that target both epithelioid and sarcomatoid (a particularly recalcitrant form) mesothelioma. Given that mesothelioma is a major occupation-related illness with no treatment to date, and little is known about mesothelioma cell surface antigens expressed by all subtypes of the disease, identification of mesothelioma antigens targeted by our panel of internalizing antibodies addresses a critical need and is likely to have a significant impact on the development of antibody-based therapeutics and diagnostics against mesothelioma.

描述（由申请人提供）：本提案的目的是确定一组内化间皮瘤细胞表面抗原的分子身份，这些抗原与一组针对上皮样和肉瘤样（一种特别顽固的形式）间皮瘤的新型内化人单链抗体（scFv）结合。 拟议的研究建立在我们最近的工作基础上，我们在活间皮瘤细胞上选择了一个组合人抗体库，并鉴定了一组内化scFv，它们原位与间皮瘤细胞的所有亚型结合，但不与正常间皮结合，并在体外介导小分子有效负载向上皮样和肉瘤样间皮瘤细胞的有效细胞内递送。我们假设这些 scFv 定义了新型间皮瘤抗原，其特异性、亚型覆盖范围和治疗潜力比目前已知的抗原明显更高。这些新型间皮瘤抗原的鉴定将（1）允许对先导抗体进行进一步工程设计，以提高亲和力、特异性、药代动力学和生物分布，（2）允许开发针对同一抗原的非重叠表位的其他抗体，这通常是诊断测定开发所必需的，（3）增强我们对涉及细胞膜的肿瘤行为的理解，这可能导致 确定进一步治疗开发的其他新靶标，(4) 为临床试验设计提供合理的理由，例如根据靶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方法来识别 scFv 靶向的肿瘤抗原。该提案的关键是我们新开发的基于人类蛋白质组的真核细胞表面展示的新型抗原识别策略。此外，我们还与博士建立了功能合作。 Burlingame 和 Chalkley 及其位于 UCSF 的国家质谱设施可鉴定难以通过表达克隆方法鉴定的肿瘤抗原，例如翻译后修饰抗原。我们建议使用我们开发和调整的方法和策略来系统地识别我们的内化人类 scFv 组靶向的间皮瘤细胞表面抗原。 公共健康相关性：本提案的目的是确定一组间皮瘤细胞表面抗原的分子身份，这些抗原与一组针对上皮样和肉瘤样（一种特别顽固的形式）间皮瘤的新型内化人单链抗体结合。鉴于间皮瘤是一种主要的职业相关疾病，迄今为止尚无治疗方法，并且对该疾病所有亚型表达的间皮瘤细胞表面抗原知之甚少，因此我们的内化抗体组针对的间皮瘤抗原的鉴定满足了关键需求，并且可能对基于抗体的间皮瘤治疗和诊断的开发产生重大影响。