Selection of internalizing human antibodies targeting pancreatic tumor cells in s

靶向胰腺肿瘤细胞的内化人抗体的选择

• 批准号: 7660256
• 负责人: BIN LIU
• 金额: $ 20.39万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-02-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7660256
• 关键词: Address; Affinity; Antibodies; Antigens; Area; Bacteriophages; Binding; Carbohydrates; Carcinoma in Situ; Cell Line; Cell surface; Cells; Chemistry; Clinical; Complex; Detection; Development; Diagnostic Neoplasm Staging; Disease; Early Diagnosis; Epitopes; Future; Genomics; Growth; Human; Image; Immunologic Surveillance; In Situ; In Vitro; Libraries; Methodology; Methods; Molecular Conformation; Monoclonal Antibodies; Morphologic artifacts; Neoplasm Metastasis; Neoplasms; Outcome; PC3 cell line; Phage Display; Post-Translational Protein Processing; Property; Proteins; Research; Scheme; Signal Transduction; Specificity; Specimen; Staging; Structure; Surface; Surface Antigens; Techniques; Tissues; Tumor Antigens; Tumor Biology; Tumor Cell Line; Tumor Pathology; Tumor Tissue; Tumor stage; Variant; anticancer research; base; design; disease diagnosis; human monoclonal antibodies; improved; laser capture microdissection; mRNA Expression; member; molecular imaging; neoplastic; neoplastic cell; novel; pancreatic neoplasm; public health relevance; receptor mediated endocytosis; research study; therapeutic target; tumor; tumor progression

DESCRIPTION (provided by applicant): The objective of this proposal is to identify internalizing human single chain antibodies (scFvs) that target pancreatic tumor cells in situ, with emphasis on (1) early stage tumors (stages 0/I), for which internalizing scFvs can be used in non-invasive imaging for early detection, and (2) late stage tumors (stages III/IV), for which internalizing scFvs can be used to develop targeted therapy based on intracellular delivery strategies. Currently, there are very few human monoclonal antibodies that target pancreatic tumors, and even fewer that specifically detect early stage pancreatic tumor to allow non-invasive imaging of asymptomatic neoplasia, an area that has a huge impact on treatment options and outcomes. Thus our proposed research would address a critical need of this field. We hypothesize that like other tumors, pancreatic tumors possess unique cell surface epitope profiles that distinguish tumors from non-neoplastic tissues. Some of these epitopes may be present early in tumor progression, allowing early detection of asymptomatic neoplasia. We further hypothesize that a subset of these tumor cell surface epitopes are internalizing, and thus can be utilized to deliver payloads intracellularly to pancreatic tumor cells. Because the tumor cell surface epitope space is composed of complex antigenic determinants including posttranslational modifications, we have taken an antibody-based approach to study the tumor cell surface. We have previously developed and utilized a naive phage antibody library containing 500 million members to identify human single chain antibodies (scFvs) targeting tumor associated internalizing epitopes, facilitating further development of targeted therapy based on intracellular delivery strategies. Recognizing that tumor cell lines often express a different cell surface antigenic profile than that of tumor cells in situ, we have recently developed a novel method that utilizes laser capture microdissection (LCM) to select phage antibody libraries on tumor cells in situ, thereby generating antibodies against clinically represented tumor antigens as opposed to possible artifacts associated with cell lines cultured in vitro. By using these techniques we are able to identify human monoclonal antibodies that target tumor antigens expressed in situ in clinical specimens, and possess novel, therapeutically useful functions such as targeted intracellular payload delivery. The precise procurement of disease cells by LCM allows tumor cells at different stages to be targeted for phage antibody selection, offering opportunities to identify stage-associated antibodies that may be used in early tumor detection. We propose to apply this strategy to select internalizing human scFvs that target pancreatic tumor cells in situ at different stages. These novel antibodies, which are human in sequence, can in the future be used to develop targeted therapeutics and non-invasive imaging-based early tumor detection. PUBLIC HEALTH RELEVANCE: This project aims to identify by laser capture microdissection internalizing human single chain antibodies that target pancreatic tumor cells in situ. These internalizing scFvs can be used to develop non-invasive imaging strategies that allow sensitive and accurate early tumor detection, and targeted therapies based on intracellular delivery strategies.

描述（由申请人提供）：该提案的目的是确定靶向胰腺肿瘤细胞的内部化人类单链抗体（SCFV），重点是（1）（1）（1）早期阶段肿瘤（0/I阶段），为此，内部化的SCFV可以用于非抗活性成像的晚期toectiv scftection II/2）（2）II（2）II（2）（2）（2）（2）（2）（2）（2）（2）（2）（2）（2）（2）（2）（2）用于基于细胞内递送策略开发有针对性的治疗。当前，靶向胰腺肿瘤的人类单克隆抗体很少，而专门检测早期胰腺肿瘤的人甚至更少，可以允许无症状的肿瘤性肿瘤性肿瘤进行非侵入性成像，该地区对治疗选择和结果产生巨大影响。因此，我们提出的研究将解决该领域的迫切需求。我们假设像其他肿瘤一样，胰腺肿瘤具有独特的细胞表面表位谱，将肿瘤与非肿瘤组织区分开。这些表位中的一些可能存在于肿瘤进展的早期，从而可以尽早发现无症状的肿瘤。我们进一步假设这些肿瘤细胞表面表位的子集在内部化，因此可以用来细胞内提供有效载荷到胰腺肿瘤细胞。由于肿瘤细胞表面表位空间由复杂的抗原决定因素（包括翻译后修饰）组成，因此我们采取了一种基于抗体的方法来研究肿瘤细胞表面。我们以前已经开发并利用了包含5亿个成员的幼稚噬菌体抗体文库，以鉴定针对肿瘤内部化表位的人类单链抗体（SCFV），从而促进了基于细胞内递送策略的进一步开发目标治疗。认识到肿瘤细胞系经常表达与原位肿瘤细胞的细胞表面抗原性谱，我们最近开发了一种新型方法，该方法利用激光捕获的微分辨率（LCM）在原位选择噬菌体抗体库，从而在肿瘤细胞原位选择抗体，从而产生与临床抗原与可能与可能与细胞相关的抗体抗体的抗体，从而培养了与细胞相关的抗菌作用，从而养成了细胞的养殖。通过使用这些技术，我们能够鉴定人类单克隆抗体，这些抗体靶向以临床样本表达的肿瘤抗原，并具有新颖的治疗功能，例如靶向细胞内有效载荷递送。 LCM对疾病细胞的精确采购允许在不同阶段将肿瘤细胞用于噬菌体选择，从而提供了鉴定与早期肿瘤检测中可能使用的阶段相关抗体的机会。我们建议将此策略应用于在不同阶段靶向胰腺肿瘤细胞的内部化体SCFV。这些新型抗体是人类的，将来可以用来开发靶向的治疗剂和非侵入性成像的早期肿瘤检测。 公共卫生相关性：该项目旨在通过激光捕获微异化的内部化，将人类的单链抗体靶向靶向胰腺肿瘤细胞的原位。这些内部化的SCFV可用于制定非侵入性成像策略，以允许敏感，准确的早期肿瘤检测，并基于细胞内递送策略的靶向疗法。