Characterization of CEACAM1 as a novel therapeutic in mantle cell lymphoma

CEACAM1 作为套细胞淋巴瘤新疗法的表征

• 批准号: 10059218
• 负责人: VU Nguyen NGO
• 金额: $ 8.23万
• 依托单位: BECKMAN RESEARCH INSTITUTE/CITY OF HOPE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-01 至 2021-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10059218
• 关键词: Accounting; Affect; Agammaglobulinaemia tyrosine kinase; Alternative Splicing; Antigen Receptors; B lymphoid malignancy; B-Cell Antigen Receptor; B-Cell Lymphomas; B-Cell Neoplasm; B-Lymphocytes; Biological Process; CEACAM1; CRISPR screen; Cell Line; Clinical Data; Cytoplasmic Tail; Data; Development; Disease; Drug resistance; Epithelial Cells; FDA approved; Future; Gene Expression Profiling; Genes; Genomic approach; Health; Hematopoietic Neoplasms; Human; Hyperactivity; ITIM; Immunotherapeutic agent; Immunotherapy; In Vitro; Integral Membrane Protein; Knowledge; Lead; Leukocytes; Lymphoma; Lymphoma cell; Malignant Neoplasms; Mantle Cell Lymphoma; Mediating; Messenger RNA; Mission; Non-Hodgkin' s Lymphoma; Oncogenic; Outcome; Pathogenesis; Patients; Pharmaceutical Preparations; Phosphorylation; Play; Prognosis; Protein Isoforms; Proteins; Public Health; Receptor Signaling; Recruitment Activity; Relapse; Research; Resistance; Role; SYK gene; Screening Result; Signal Transduction; Signaling Molecule; T-Lymphocyte; Testing; United States National Institutes of Health; carcinoembryonic antigen-related cell adhesion molecules; chemotherapy; chimeric antigen receptor; chimeric antigen receptor T cells; effective therapy; functional genomics; genome-wide; improved; in vivo; negative affect; novel; novel therapeutics; overexpression; patient derived xenograft model; pre-clinical; therapeutic target; tumor

ABSTRACT Mantle cell lymphoma (MCL), accounting for 6-8% of all non-Hodgkin lymphomas (NHLs), is an aggressive and difficult-to-treat B-cell malignancy. MCL patients have a dismal prognosis with a median overall survival of only 3-5 years owing to frequent relapse and resistance to contemporary chemotherapy. Despite recent treatment advances with the FDA-approved drug ibrutinib, which targets the B-cell receptor (BCR) signaling molecule Bruton’s tyrosine kinase (BTK), one-third of MCL patients are ibrutinib-resistant, and even ibrutinib-sensitive patients eventually acquire resistance to the drug. Patients who are insensitive or progress during ibrutinib treatment have very poor outcomes. Thus, more effective post-ibrutinib treatments are urgently needed. Abnormal BCR signaling plays a key oncogenic signal in MCL, but the underlying mechanism is not known. Using an integrated functional genomics approach combining a genome-wide CRISPR-Cas9 screen and gene expression profiling, our group has uncovered CEACAM1 (carcinoembryonic antigen-related cell adhesion molecule-1) as a potential contributor to BCR hyperactivity in MCL. CEACAM1 is a type 1 transmembrane protein expressed as different isoforms due to alternative splicing in epithelial cells and activated leukocytes. In contrast to the short isoform, the CEACAM1 long isoform possesses a cytoplasmic tail that contains the immunoreceptor tyrosine-based inhibitory motif (ITIM) known to negatively regulate the antigen receptor signaling in T cells. We found that CEACAM1 is overexpressed specifically in MCL, as compared to naïve B cells or other B-cell malignancies. Consistent with our CRISPR screen results, depletion of CEACAM1 is toxic in multiple MCL cell lines, but not in non-MCL lymphoma cells. CEACAM1 depletion also markedly reduced proximal BCR signaling (e.g., SYK phosphorylation) and BCR-mediated Ca2+ signaling. More importantly, using T cells that express the CEACAM1-specific chimeric antigen receptor (CAR), we showed that these CAR T cells could effectively eliminate MCL tumors while sparing non-MCL lymphoma in vitro. On the basis of these findings, we hypothesize that CEACAM1 is essential for the oncogenic BCR signaling in MCL and thus a valid therapeutic target for this malignancy. We will test the hypothesis through accomplishing the following specific aims: 1) Determine the mechanisms underlying CEACAM1-regulated BCR signaling; 2) Characterize the MCL-specific expression of CEACAM1; 3) Develop a CAR T-cell strategy to treat MCL in vivo using pre-clinical patient-derived xenograft (PDX) models. Findings from this proposal will improve our limited knowledge of the role of CEACAM1 in MCL pathogenesis. We anticipate that successful completion of the project will provide relevant pre-clinical data for future studies towards developing a novel, effective therapy for MCL.

摘要 套细胞淋巴瘤(MCL)占所有非霍奇金淋巴瘤(NHL)的6-8%，是一种侵袭性和 难以治疗的B细胞恶性肿瘤。MCL患者的预后很差，总体存活率中位数仅为 3-5年，因经常复发和对现代化疗耐药。尽管最近接受了治疗 FDA批准的针对B细胞受体(BCR)信号分子的药物伊布鲁替尼的研究进展 布鲁顿酪氨酸激酶(BTK)，三分之一的MCL患者对伊布鲁替尼耐药，甚至对伊布鲁替尼敏感 患者最终会对这种药物产生抗药性。在伊布鲁替尼期间不敏感或进展的患者 治疗的结果非常糟糕。因此，迫切需要更有效的伊布鲁替尼后治疗。 异常的BCR信号在MCL中起着关键的致癌信号作用，但其潜在的机制尚不清楚。 使用结合全基因组CRISPR-Cas9屏幕和基因的集成功能基因组学方法 表达谱，我们的团队发现了CEACAM1(癌胚抗原相关细胞黏附 分子-1)作为MCL中BCR过度活动的潜在贡献者。CEACAM1是一种1型跨膜蛋白 由于在上皮细胞和激活的白细胞中的选择性剪接，表达为不同的异构体。相比之下， 对于短的异构体，CEACAM1长的异构体有一个含有免疫受体的细胞质尾巴 酪氨酸抑制基序(ITIM)对T细胞中的抗原受体信号具有负性调节作用。我们 与单纯B细胞或其他B细胞相比，CEACAM1在MCL中高表达 恶性肿瘤。与我们的CRISPR筛查结果一致，CEACAM1的耗竭在多个MCL细胞中是有毒的 株，但在非MCL淋巴瘤细胞中不表达。CEACAM1缺失也显著减少了近端BCR信号 (例如，SYK磷酸化)和BCR介导的钙信号转导。更重要的是，使用T细胞来表达 CEACAM1特异性嵌合抗原受体(CAR)，我们发现这些CAR T细胞可以有效地 在体外消除MCL肿瘤的同时保留非MCL淋巴瘤。根据这些发现，我们假设 CEACAM1是MCL致癌BCR信号转导所必需的，因此是治疗MCL的有效靶点 恶毒。我们将通过实现以下具体目标来检验假设：1)确定 CEACAM1调节BCR信号的机制；2)表征MCL特异性表达 CEACAM1；3)开发CAR T细胞策略，使用临床前患者来源的异种移植在体内治疗MCL (PDX)型号。这项提案的发现将改善我们对CEACAM1在MCL中的作用的有限了解 发病机制。我们预计该项目的成功完成将为以下项目提供相关的临床前数据 未来的研究方向是开发一种新的、有效的MCL治疗方法。