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.

抽象的 地幔细胞淋巴瘤（MCL），占所有非霍奇金淋巴瘤（NHLS）的6-8％，是一种侵略性， 难以治疗的B细胞恶性肿瘤。 MCL患者的预后很糟糕，总体生存仅为 3 - 5年由于经常缓解和对当代化学疗法的抵抗力。尽管最近的治疗方法 FDA批准的药物ibrutinib的进步，该药物靶向B细胞受体（BCR）信号分子 Bruton的酪氨酸激酶（BTK），三分之一的MCL患者是抗ibrutinib的，甚至对Ibrutinib敏感的 患者最终获得了对药物的耐药性。伊布鲁替尼期间不敏感或进步的患者 治疗的结果很差。这是迫切需要更有效的后纤维替尼治疗。 异常的BCR信号在MCL中起关键的致癌信号，但尚不清楚潜在的机制。 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 表达谱分析，我们的小组发现了CEACAM1（癌症抗原相关的细胞粘附） 分子1）是MCL中BCR多动症的潜在贡献者。 CEACAM1是1型跨膜蛋白 由于上皮细胞和活化的白细胞中的替代剪接而表示为不同的同工型。相比之下 对于短同工型，CEACAM1长同工型具有含有免疫感受器的细胞质尾巴 已知基于酪氨酸的抑制基序（ITIM）对T细胞中的抗原受体信号传导负面调节。我们 发现与幼稚的B细胞或其他B细胞相比，CEACAM1在MCL中特别表达过表达 恶性。与我们的CRISPR屏幕结果一致，CEACAM1的部署在多个MCL细胞中有毒 线，但不在非MCL淋巴瘤细胞中。 CEACAM1耗竭也显着降低了近端BCR信号传导 （例如，SYK磷酸化）和BCR介导的Ca2+信号传导。更重要的是，使用表达的T细胞 CEACAM1特异性嵌合抗原受体（CAR），我们表明这些CAR T细胞可以有效地 在体外保留非MCL淋巴瘤时消除MCL肿瘤。根据这些发现，我们假设 该CEACAM1对于MCL中的致癌BCR信号至关重要，因此是有效的治疗靶标 恶性。我们将通过完成以下特定目的来检验假设：1）确定 CEACAM1调节的BCR信号传导的机制； 2）表征MCL特异性的表达 ceacam1; 3）制定一种使用前临床患者衍生异种移植物在体内治疗MCL的汽车T细胞策略 （PDX）模型。该提案的发现将提高我们对CEACAM1在MCL中的作用的有限了解 发病。我们预计该项目的成功完成将为临床前数据提供 未来的研究，用于开发一种新颖的，有效的MCL疗法。