Immunology of Unique Tumor Specific Antigens

独特的肿瘤特异性抗原的免疫学

• 批准号: 8743177
• 负责人: Hans Schreiber
• 金额: $ 22.44万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-07-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8743177
• 关键词: Acetylgalactosamine; Address; Adoptive Immunotherapy; Adoptive Transfer; Affinity; Antibodies; Antigens; Appearance; Autologous; B lymphoid malignancy; B-Lymphocytes; Binding; Breast; CD19 gene; Cell Line; Chimeric Proteins; Clinical; Colon; Cross Presentation; Defect; Epitopes; Event; Fc Receptor; Genes; Glycopeptides; Growth; Hematologic Neoplasms; Human; Immunoglobulin G; Immunoglobulin M; Immunology; Immunotherapy; Interleukin-15; Link; Lung; MS4A1 gene; Malignant - descriptor; Malignant Neoplasms; Membrane Glycoproteins; Monoclonal Antibodies; Mucin-1 Staining Method; Mus; Mutation; Natural Killer Cells; Normal tissue morphology; Ovary; Pancreas; Patients; Peptides; Primary Neoplasm; Prostate; Proteins; Relapse; Serine; Solid; Solid Neoplasm; Specificity; T-Cell Activation; T-Independent Antigens; T-Lymphocyte; Testing; Threonine; Toxic effect; Transgenic Mice; Trees; Tumor Antigens; Tumor Escape; Viral Tumor Antigens; base; cancer cell; cancer immunotherapy; chimeric antibody; clinically relevant; design; glycosylation; mouse model; mutant; prevent; protein aminoacid sequence; prototype; public health relevance; receptor; success; sugar; tumor

DESCRIPTION (provided by applicant): The objective is to examine Tn-O-glycopeptide antigens as targets for immunotherapy in a range of human cancers. A large percentage of common human cancers (e.g., colon, breast, lung, prostate, ovary and pancreas) have defects in O-linked glycosylation thereby generating tumor-specific epitopes on surface glycoproteins of cancer cells. The defect is a truncation of the growing sugar tree after linkage of the first sugar N-acetylgalactosamine (GalNAc), to threonine or serine on the peptide chain of proteins by an ?1-O-glycosyl linkage. The linked single sugar is referred to as Tn (T antigen nouvelle) and can be detected by "anti-Tn" IgM antibodies. The cancer-specific truncation can also be detected by high-affinity "anti-Tn-O-glycopeptide" IgG antibodies. They, in contrast, engulf the single sugar moiety while recognizing surrounding specific peptide sequences. Such IgG antibodies react with all cancers that have the required glycosylation defect and provide the same aberrantly glycosylated surface glycoprotein. Three such monoclonal antibodies, 237, 5E5 and 3H4, have been cloned and sequenced to generate (i) chimeric antibody receptors (CARs) for transduction into T cells for adoptive transfer and (ii) bispecific fusion proteins to engage the host's NK and cells in the margins of large solid tumors. The fusion proteins will consist of the binding domain and IL15 linked to IL15R?. Aim 1 is to determine the potential of 237 CAR-transduced T cells targeting a tumor-specific Tn-O-glycopeptide epitope to eradicate solid non-hematopoietic tumors in a syngeneic mouse model. It will be determined whether these T cells eradicate clinical-size solid tumors without toxicity to normal tissues, by what mechanism destruction or escape occurs, whether the sequential events leading to destruction differ from those caused by TCR transduced T cells, and whether T cells transduced with CARs can destroy cancer cells that micro-disseminate from solid primary tumors. Aim 2 is to determine the potential of fusion proteins carrying IL15-IL15R? and having 237 or 5E5 receptors to target tumor-specific Tn-O-glycopeptide epitopes. It will be determined whether fusion proteins carrying the anti-Tn- O-glycopeptide scFv and IL15-IL15R? can cause the appearance of densely granulated NK cells at the tumor margin and rescue tolerant intratumoral T cells to reject large established tumors. Aim 3 is to generate and evaluate CARs recognizing Tn-O-glycopeptide epitopes on the deglycosylated form of MUC1 found in human cancers. It will be tested whether such CARs lack toxicity towards normal tissues expressing the normal, fully glycosylated form of MUC1 in MUC1-transgenic mice while eliminating clinical size tumors. Furthermore, a panel of isogenic cell lines of common human cancers, all lacking Cosmc function (leading to defects in O-linked glycosylation), will be used to induce and select for additional cancer-specific antibodies recognizing Tn-O-glycopeptide epitopes on other surface glycoproteins. With these antibodies, we will make additional CARs (and fusions proteins) to be used in combination with 5E5 to prevent tumor escape.

描述（由申请人提供）：目的是检查TN-O-糖肽抗原作为一系列人类癌症的免疫疗法的靶标。大部分的普通癌症（例如结肠，乳腺癌，肺，前列腺，卵巢和胰腺）在O连接的糖基化中存在缺陷，从而在癌细胞表面糖蛋白上产生肿瘤特异性表位。缺陷是在第一糖N-乙酰乳糖苷（GalNAC）连接后生长的糖树的截断，通过？1-O-糖基链接在蛋白质肽链上与苏氨酸或丝氨酸的截断。链接的单糖称为TN（t抗原Nouvelle），可以通过“抗TN” IGM抗体检测。癌症特异性截断也可以通过高亲和力的“抗TN-O-糖肽” IgG抗体检测到。相比之下，它们吞噬了单个糖部分，同时识别周围的特定肽序列。这种IgG抗体与所有具有所需的糖基化缺陷并提供相同异常的糖基化表面糖蛋白的癌症反应。三种这样的单克隆抗体，237、5E5和3H4，已被克隆并测序，以产​​生（i）嵌合抗体受体（CAR），用于转导向T细胞，以进行收养和（ii）双特异性融合蛋白使宿主的NK和细胞与大型固体晶状体的NK和细胞吸引。融合蛋白将由与IL15R相关的结合域和IL15组成。目的1是确定针对肿瘤特异性TN-O-糖肽表位的237个CAR转导的T细胞的潜力，以消除合成小鼠模型中的固体非造成肿瘤。将确定这些T细胞是否通过发生什么机制或逃逸发生了哪些机制或逃生的临床大小的实体瘤，是否发生了导致破坏的顺序事件与TCR转导的T细胞引起的顺序事件，以及用CARS转导的T细胞可以破坏从固体原发性肿瘤中微异常的癌细胞破坏癌细胞。 AIM 2是确定携带IL15-IL15R的融合蛋白的潜力？并具有237或5E5受体靶向肿瘤特异性TN-O-糖肽表位。将确定携带抗TN-O-糖肽SCFV和IL15-IL15R的融合蛋白是否？会导致在肿瘤边缘的密集粒细胞的出现，并挽救耐肿瘤内T细胞以拒绝大型已建立的肿瘤。 AIM 3是生成和评估识别人类癌症中MUC1脱脂形式的TN-O-糖肽表位的汽车。将测试此类汽车是否对表达正常的，完全糖基化的MUC1在MUC1-转基因小鼠中表达正常，完全糖基化形式的毒性，同时消除临床大小的肿瘤。此外，所有缺乏COSMC功能（导致O连接糖基化的缺陷）的一组同源细胞系将用于诱导并选择识别其他表面糖蛋白上TN-O-糖肽表位的其他癌症特异性抗体。使用这些抗体，我们将与5E5结合使用其他汽车（和融合蛋白），以防止肿瘤逸出。