Transmembrane Proteins Involved in Human Tumor Expansion

参与人类肿瘤扩张的跨膜蛋白

• 批准号: 8396608
• 负责人: JAMES P QUIGLEY
• 金额: $ 13.96万
• 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/8396608
• 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; Health; 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; Screening procedure; 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; 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. PUBLIC HEALTH RELEVANCE: Deaths from cancer occur mainly because tumor cells spread from the primary tumor site to other vital organs and tissues. In order for tumor cells to escape from the primary tumor and travel to distant organs, they produce different levels of functioning proteins. The goal of the proposed research is to use a modified system of monoclonal antibody generation in order to identify the relevant proteins that contribute to tumor cell spread. Oncologists could target those critical protein molecules and might also employ the specific antibodies as therapeutic agents.

描述（由申请人提供）：为了确定在功能上有助于人类肿瘤细胞转移的特定蛋白质抗原，我们的实验室启动了减法免疫方法，以提高抑制人类肿瘤传播的独特单克隆抗体（MAB）。对产生的mAb的面板进行了筛选，以抑制鸡胚胎和转移小鼠模型中人类肿瘤传播的能力。使用这种方法，我们生成了独特的抗体。其中一个MAB 1A5识别四跨果蛋白家族的特定成员CD151，并相应地抑制了> 90％的转移。另一种减去免疫抗体MAB 41-2识别肿瘤细胞跨膜蛋白CDCP1/SIMA-135。这种MAB 41-2抑制了总体转移60-80％，但是MAB 41-2及其靶抗原的机制尚不清楚。因此，该更新的建议目标是阐明MAB 41-2的转移抑制机制，并对其同源抗原CDCP1进行结构函数分析，以确定CDCP1在癌症传播中的贡献作用。将mAb 41-2的定时添加到动物中，然后定量测量肿瘤细胞内血管内停滞，凋亡，生存，渗出和次级灶的建立将确定何时，何时何地和如何阻断恶性功能。这些方法将在肿瘤传播的小鼠模型中扩展和确认，包括原位植入方法。 MAB 41-2的抗原Cdcp1作为生存因子的作用将通过CDCP1的细胞质结构域和Cdcp1的细胞外幼崽结构域进行突变分析进行测试。将使用组合文库的使用来识别CDCP1的未知天然配体，并生成与CDCP1的新型抗体。这种更新的另一个目的是表征其他成对的人类肿瘤变体，这些变体在肿瘤形成能力上相似，但在其转移特性上有很大差异，并在新的减法免疫方法中使用这些细胞对来产生新的功能阻断抗体。 MAB的面板将在我们已建立的模型中筛选出差异反应性和功能阻滞性能。我们还建议启动潜在的翻译方法并生成新的转移阻滞，单链可变片段（SC FV）人源化抗体，这些抗体在癌症类型特异性的原位小鼠模型中有效，并且可以代表可能靶向肿瘤疗法的独特试剂。公共卫生相关性：癌症死亡的原因主要是因为肿瘤细胞从原发性肿瘤部位扩散到其他重要器官和组织。为了使肿瘤细胞从原发性肿瘤中逸出并传播到远处的器官，它们会产生不同水平的功能蛋白。拟议的研究的目的是使用修饰的单克隆抗体产生系统，以鉴定有助于肿瘤细胞扩散的相关蛋白质。肿瘤学家可以靶向那些关键的蛋白质分子，还可以使用特定的抗体作为治疗剂。