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: 10470672
• 负责人: John Zhong
• 金额: $ 37.84万
• 依托单位: LOMA LINDA UNIVERSITY
• 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 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10470672
• 关键词: Molecular; Characteristics; Treatment; resistant; risk

 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（申请人提供）：本项目的目的是利用单细胞技术鉴定高危神经母细胞瘤患者多药耐药的分子特征。我们提出了一种范式转换的方法来映射到一个单一的细胞突变，并确定单个细胞的分子相似性的亚克隆。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），使用下一代测序（NGS）识别多药耐药突变。这些NGS鉴定的突变将被定位到具有靶向单细胞RNA-seq的单个细胞中，以显示这些突变存在于克隆（一组单细胞）中，并且克隆在治疗期间扩增。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社区。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（通常具有实验偏倚）进行表征，从而改变了肿瘤分子表征的范式。这些结果对目前神经母细胞瘤的治疗有直接和直接的影响。耐药克隆的分子谱可以开发成敏感的PCR检测，用于在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（在化疗期间具有特定亚克隆）的治疗耐药性。我们在这项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。