Molecular Characteristics of Treatment-resistant High-risk Neuroblastoma

难治性高危神经母细胞瘤的分子特征

• 批准号: 9247150
• 负责人: John Zhong
• 金额: $ 37.84万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9247150
• 关键词: Address; Adult; Alpha Cell; Biological Assay; Biological Markers; Bone Marrow; Cell Line; Cells; Characteristics; Child; Childhood; Communities; DNA Sequence Alteration; Derivation procedure; Diagnosis; Disease; Drug resistance; Drug resistance pathway; Early Diagnosis; Foundations; Frequencies; Genes; Genetic; Goals; Individual; Malignant Neoplasms; Maps; Microfluidics; Minority; Molecular; Molecular Cloning; Molecular Profiling; Multi-Drug Resistance; Mutation; Neuroblastoma; Pathway interactions; Patients; Refractory; Relapse; Research; Residual Neoplasm; Resistance; Resistance profile; Sampling; Selection for Treatments; Solid Neoplasm; Testing; Tumor Cell Line; Variant; base; chemotherapy; childhood cancer mortality; cytotoxicity; disease diagnosis; drug development; high risk; improved outcome; neuroblastoma cell; next generation sequencing; novel therapeutic intervention; novel therapeutics; personalized medicine; public health relevance; resistance mechanism; single cell technology; stable cell line; therapy resistant; transcriptome sequencing; tumor

 DESCRIPTION (provided by applicant): The goal of this project is to identify molecular characteristics of multi-drug resistance in high-risk neuroblastoma patients with single-cell technology. We propose a paradigm-shift approach to map mutations into a single cell and identify sub-clones by molecular similarity of individual cells. Neuroblastoma is the common solid tumor in children, and it accounts for 15% of deaths from cancer in children. Here, we propose to identify multidrug-resistant mutations with Next Generation Sequencing (NGS) based on mutation frequency changes (from minority to majority) due to treatment selection. These NGS-identified mutations will be localized into individual cells with targeted single-cell RNA-seq to show that these mutations exist in a clone (a group of single-cells) and the clones expand during treatment. Finally, the resistant sub-clones will be isolated or generated for functional assays and distributed to research community. This approach shifts the paradigm of tumor molecular characterization by clustering molecularly similar cells into a clone rather than isolating a clone (often with experimental bias) for characterization. The results have direct and immediate impacts on current treatment of neuroblastoma. The molecular profiles of resistant clones can be developed into sensitive PCR assays for early detection of drug-resistance at diagnosis, and for personalized therapies to overcome treatment resistance at specific patients (who have specific sub-clones during chemotherapy). The single-cell approach that we develop in this proposal would be directly relevant to studies of other childhood and adult cancers as well.

 描述（由申请人提供）：该项目的目标是利用单细胞技术鉴定高危神经母细胞瘤患者多重耐药的分子特征。我们提出了一种范式转换方法，将突变映射到单个细胞中，并通过单个细胞的分子相似性来识别亚克隆。神经母细胞瘤是儿童常见的实体瘤，占儿童癌症死亡人数的 15%。在这里，我们建议根据治疗选择导致的突变频率变化（从少数到多数），通过下一代测序（NGS）来识别多重耐药突变。这些 NGS 识别的突变将通过靶向单细胞 RNA-seq 定位到单个细胞中，以显示这些突变存在于克隆（一组单细胞）中，并且克隆在治疗过程中会扩展。最后，将分离或产生抗性亚克隆用于功能测定并分发给研究界。这种方法通过将分子相似的细胞聚集成克隆而不是分离克隆（通常带有实验偏差）来进行表征，从而改变了肿瘤分子表征的范式。结果对当前神经母细胞瘤的治疗产生直接和直接的影响。耐药克隆的分子谱可以开发为敏感的 PCR 检测，用于诊断时早期检测耐药性，并用于个性化治疗以克服特定患者（化疗期间具有特定亚克隆）的治疗耐药性。我们在本提案中开发的单细胞方法也将与其他儿童和成人癌症的研究直接相关。