Transmembrane Proteins Involved in Human Tumor Expansion

参与人类肿瘤扩张的跨膜蛋白

• 批准号: 8665128
• 负责人: JAMES P QUIGLEY
• 金额: $ 3.03万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-16 至 2014-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8665128
• 关键词: Affinity Chromatography; Animals; Antibodies; Antigen Targeting; Antigens; Apoptosis; Binding; Blocking Antibodies; Blood; Cancer Patient; Cell Surface Proteins; Cells; Cessation of life; Chick Embryo; Co-Immunoprecipitations; Complementary DNA; Cysteine; Cytoplasmic Tail; Distant; Epitopes; Extravasation; Family; Generations; Glycoproteins; Goals; Grant; Human; Immunization; Integral Membrane Protein; Laboratories; Libraries; Ligands; Malignant - descriptor; Malignant Neoplasms; Measurement; Microscopy; Modeling; Monitor; Monoclonal Antibodies; Mus; Neoplasm Metastasis; Oncologist; Organ; Peptide Library; Primary Neoplasm; Property; Proteins; Proteomics; Reagent; Research; Role; Site; Stem cells; Structure; System; Testing; Therapeutic Agents; Time; Tissues; Tolerogen; Travel; Tumor Expansion; Tyrosine; Variant; base; cancer type; cell type; combinatorial; congenic; extracellular; human PHEMX protein; humanized antibody; implantation; in vivo; in vivo Model; member; mouse model; neoplastic cell; novel; protein function; research study; screening; therapeutic target; tool; translational approach; tumor; tumor progression

DESCRIPTION (provided by applicant): To identify specific protein antigens that functionally contribute to human tumor cell metastasis, our laboratory initiated subtractive immunization approaches to raise unique monoclonal antibodies (mAbs) that inhibit human tumor dissemination. Panels of generated mAbs are screened for their ability to inhibit human tumor dissemination in chick embryo and mouse models of metastasis. Using this approach, we have generated unique antibodies. One of them, mAb 1A5, recognizes a specific member of the tetraspanin family, CD151, and correspondingly inhibits metastasis by > 90%. Another subtractive immunization antibody, mAb 41-2, recognizes a tumor cell transmembrane protein, CDCP1/SIMA-135. This mAb 41-2 inhibits overall metastasis by 60-80%, however the mechanism of mAb 41-2 and that of its target antigen are unknown. Thus, a proposed goal of this renewal is to elucidate the metastasis-inhibition mechanism of mAb 41-2 and to conduct structure- function analysis of its cognate antigen, CDCP1, in order to determine the contributory role of CDCP1 in cancer dissemination. Timed additions of mAb 41-2 into animals followed by quantitative measurements of tumor cell intravascular arrest, apoptosis, survival, extravasation and establishment of secondary foci will determine when, where and how mAb 41-2 blocks malignant function. These approaches will be expanded and confirmed in mouse models of tumor dissemination, including orthotopic implantation approaches. That mAb 41-2's antigen, CDCP1, functions as a survival factor will be tested with mutational analyses of CDCP1's cytoplasmic domain and CDCP1's extracellular CUB domains. The use of combinatorial libraries will be employed to identify unknown natural ligand(s) for CDCP1 and to generate new distinctive antibodies to CDCP1. Another aim of this renewal is to characterize additional pairs of congenic human tumor variants that are similar in tumor-forming ability but differ substantially in their metastasis properties, and to use these cell pairs in new subtractive immunization approaches to generate novel function-blocking antibodies. Panels of mAbs will be screened for differential reactivity and function-blocking properties in our established models. We also propose to initiate potential translational approaches and generate new metastasis-blocking, single chain variable fragment (sc Fv) humanized antibodies that are effective in cancer type-specific orthotopic mouse models and could represent unique reagents for possible targeted tumor therapy.

描述（由申请人提供）：为了鉴定功能上有助于人类肿瘤细胞转移的特异性蛋白抗原，我们的实验室启动了消减免疫方法，以产生抑制人类肿瘤传播的独特单克隆抗体（mAb）。筛选产生的mAb组在鸡胚和小鼠转移模型中抑制人肿瘤播散的能力。使用这种方法，我们已经产生了独特的抗体。其中之一，mAb 1A 5，识别四跨膜蛋白家族的特定成员，CD 151，并相应地抑制转移> 90%。另一种消减免疫抗体mAb 41-2识别肿瘤细胞跨膜蛋白CDCP 1/SIMA-135。该mAb 41-2抑制了60- 80%的总转移，然而mAb 41-2及其靶抗原的机制尚不清楚。因此，该更新的拟定目标是阐明mAb 41-2的转移抑制机制，并对其同源抗原CDCP 1进行结构-功能分析，以确定CDCP 1在癌症播散中的贡献作用。将mAb 41-2定时添加到动物中，然后定量测量肿瘤细胞血管内停滞、凋亡、存活、外渗和继发病灶的建立，将确定mAb 41-2何时、何地和如何阻断恶性功能。这些方法将在肿瘤播散的小鼠模型中得到扩展和证实，包括原位植入方法。mAb 41-2的抗原CDCP 1作为存活因子的功能将通过CDCP 1的胞质结构域和CDCP 1的胞外CUB结构域的突变分析进行测试。将采用组合文库的使用来鉴定CDCP 1的未知天然配体并产生针对CDCP 1的新的独特抗体。这种更新的另一个目的是表征在肿瘤形成能力上相似但在其转移特性上显著不同的其他同类人类肿瘤变体对，并在新的消减免疫方法中使用这些细胞对以产生新的功能阻断抗体。将在我们建立的模型中筛选mAb组的差异反应性和功能阻断特性。我们还提出启动潜在的翻译方法，并产生新的转移阻断，单链可变片段（sc Fv）的人源化抗体，是有效的癌症类型特异性原位小鼠模型，并可能代表独特的试剂可能有针对性的肿瘤治疗。