Defining the mechanisms of B7-H3 overexpression and role in neuroblastoma metastasis and immune evasion

定义 B7-H3 过度表达的机制及其在神经母细胞瘤转移和免疫逃避中的作用

• 批准号: 10750399
• 负责人: Catherine Leona-Grace Wingrove
• 金额: $ 4.77万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10750399
• 关键词: 3-Dimensional; Adrenergic Agents; Adult; Antibody-drug conjugates; Antigens; Automobile Driving; Binding; Biological Assay; Biological Process; C57BL/6 Mouse; CD276 gene; CD3 Antigens; CRISPR/Cas technology; Cell Line; Cell Proliferation; Cell Surface Proteins; Cell physiology; Cell secretion; ChIP-seq; Characteristics; Chemotherapy-Oncologic Procedure; Childhood; Childhood Extracranial Solid Tumor; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Credentialing; Cytolysis; Data; Dependence; Development; Diagnosis; Effector Cell; Event; FDA approved; Family; Fiber; Future; Genes; Genetic; Genetic Transcription; Growth; Human; Human Cell Line; Immune; Immune Evasion; Immunocompetent; Immunologic Deficiency Syndromes; Immunophenotyping; Immunotherapeutic agent; Immunotherapy; In Vitro; Inflammatory; Integral Membrane Protein; Interferons; Interleukin-2; International; Invaded; Life; Ligands; Luciferases; MART-1 Tumor Antigen; MYCN gene; Malignant Childhood Neoplasm; Malignant Neoplasms; Mediating; Mediator; Mesenchymal; Migration Assay; Monoclonal Antibody Therapy; Morbidity - disease rate; Mutate; Mutation; National Research Service Awards; Natural Killer Cell Immunotherapy; Natural Killer Cells; Neoplasm Metastasis; Neuroblastoma; Oncogenes; Oncogenic; Oncoproteins; Operative Surgical Procedures; Pain; Pathway interactions; Patient-Focused Outcomes; Patients; Proliferating; Proteins; Proto-Oncogene Proteins c-myc; Public Health; Radiation; Recombinants; Research; Resistance; Role; Scientist; Seminal; Solid; Solid Neoplasm; Survival Rate; Survivors; Sympathetic Nervous System; System; T cell response; T-Cell Activation; T-Lymphocyte; TNF gene; Testing; Toxic effect; Transforming Growth Factor beta; Transgenic Organisms; Translational Research; Treatment Protocols; Tumor Antigens; Work; Xenograft procedure; antigen-specific T cells; career; cell growth; chemoradiation; chimeric antigen receptor T cells; clinically relevant; comorbidity; cytokine; cytotoxicity; differential expression; disorder risk; evidence base; high risk; immunoregulation; improved; improved outcome; in vivo; inhibitor; knock-down; migration; mortality; mouse model; neuroblastoma cell; new therapeutic target; novel; novel therapeutics; overexpression; pharmacologic; pre-clinical; promoter; research clinical testing; response; side effect; skills; success; targeted treatment; therapeutic target; transcription factor; treatment strategy; tumor; tumor growth; tumor microenvironment; wound

Project Summary Neuroblastoma is a cancer derived from the developing sympathetic nervous system and is the most commonly diagnosed extracranial solid tumor of childhood. Despite an intensive treatment regimen of chemotherapy, surgery, radiation, and immunotherapy, the five-year survival rate of high-risk neuroblastoma patients remains at only 50%. The low tumor mutational burden of neuroblastoma has challenged the development of targeted- and immuno-therapies, however moderate success has been achieved by targeting GD2 with monoclonal antibody therapy, credentialing immunotherapeutic treatment strategies for neuroblastoma as a promising approach. The success of GD2-targeted therapy to date has been hindered due to debilitating side effects from on-target/off-tumor toxicity since GD2 is also expressed on pain fibers, and antigen loss as a mechanism of therapy resistance. Therefore, there is an unmet need for the discovery of new therapeutic targets in neuroblastoma. B7-H3, encoded by the CD276 gene, is a type 1 transmembrane protein in the B7 family of immunoregulatory proteins and is highly expressed in many adult and pediatric cancers, including neuroblastoma. In addition to being implicated in immunoinhibition, B7-H3 may also mediate tumor migration and metastasis. Preclinical success of several immunotherapeutic strategies directed toward B7-H3, including CAR-T cells and antibody drug conjugates, suggest that B7-H3 is a targetable tumor-associated antigen with several pediatric clinical trials ongoing or planned. Therefore, it is critical to understand the oncogenic functions of B7-H3 and how its expression is regulated to anticipate mechanisms of therapy resistance. Our central hypothesis is that B7-H3 promotes neuroblastoma metastasis and immune evasion, and its expression is regulated by tumor microenvironment-derived cytokines and neuroblastoma-specific transcription factors. Our preliminary data shows that B7-H3 knockdown using CRISPRi in neuroblastoma cell lines inhibits cellular proliferation. Additionally, ChIP-sequencing data identifies regions of MYC and MYCN binding at the B7-H3 promoter indicating a potential role of the MYC transcription factors in regulating B7-H3 expression. B7-H3 expression may also be regulated by inflammatory cytokines, as neuroblastoma cell lines upregulate B7-H3 expression following TNF-α or TGF-β exposure. Finally, recombinant human B7-H3 inhibits T cell activation, TNF-α, and IFN-y. Uncovering how B7-H3 promotes immune evasion in neuroblastoma is crucial given the T and NK cell-based immunotherapies undergoing clinical testing. We propose that B7-H3 is a multifunctional protein that serves as a promising therapeutic target in neuroblastoma. This NRSA F31 will define the mechanisms of neuroblastoma dependance on B7-H3 for metastasis and immune evasion, while also defining mechanisms of overexpression, to inform future B7-H3-targeting therapies and ultimately improve outcomes for patients with high-risk neuroblastoma.

项目概要 神经母细胞瘤是一种源自发育中的交感神经系统的癌症，是最常见的癌症 诊断出儿童颅外实体瘤，尽管采用了强化化疗方案， 手术、放疗和免疫治疗，高危神经母细胞瘤患者的五年生存率仍然很高 神经母细胞瘤的低肿瘤突变负荷对靶向治疗的发展提出了挑战。 和免疫疗法，但是通过单克隆抗体靶向 GD2 已取得一定成功 抗体疗法，证明神经母细胞瘤的免疫疗法是一种有前途的治疗策略 迄今为止，GD2 靶向治疗的成功因 GD2 的副作用而受到阻碍。 由于 GD2 也在疼痛纤维上表达，因此存在靶向/脱肿瘤毒性，并且抗原丢失是一种机制 因此，发现新的治疗靶点的需求尚未得到满足。 B7-H3 由 CD276 基因编码，是 B7 家族的 1 型跨膜蛋白。 免疫调节蛋白，在许多成人和儿童癌症中高度表达，包括 除了参与免疫抑制之外，B7-H3 还可能介导肿瘤迁移。 多种针对 B7-H3 的免疫治疗策略的临床前成功，包括 CAR-T 细胞和抗体药物偶联物表明 B7-H3 是一种可靶向的肿瘤相关抗原 一些正在进行或计划进行的儿科临床试验因此，了解致癌功能至关重要。 B7-H3 的表达以及如何调节其表达以预测治疗耐药机制。 假设B7-H3促进神经母细胞瘤转移和免疫逃避，其表达量为 受肿瘤微环境衍生的细胞因子和神经母细胞瘤特异性转录因子的调节。 初步数据表明，在神经母细胞瘤细胞系中使用 CRISPRi 敲低 B7-H3 可抑制细胞 此外，ChIP 测序数据还识别了 B7-H3 上 MYC 和 MYCN 结合的区域。 启动子表明 MYC 转录因子在调节 B7-H3 表达中的潜在作用。 表达也可能受到炎症细胞因子的调节，因为神经母细胞瘤细胞系上调 B7-H3 最后，重组人 B7-H3 抑制 T 细胞活化。 鉴于 T，揭示 B7-H3 如何促进神经母细胞瘤的免疫逃避至关重要。 我们认为 B7-H3 是一种多功能的基于 NK 细胞的免疫疗法。 NRSA F31 蛋白将成为神经母细胞瘤的一个有前途的治疗靶点。 神经母细胞瘤依赖 B7-H3 进行转移和免疫逃避的机制，同时还定义了 过度表达的机制，为未来的 B7-H3 靶向治疗提供信息并最终改善结果 高危神经母细胞瘤患者。