(PQ #4) Single cell barcoding for studies of study clonal evolution in glioblastoma

（PQ

• 批准号: 9353234
• 负责人: Erik Sulman
• 金额: $ 33.2万
• 依托单位: JACKSON LABORATORY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9353234
• 关键词: Address; Adult; Aftercare; BRAF gene; Behavior; Biological Markers; Biological Models; Biopsy; Blood - brain barrier anatomy; Brain Neoplasms; Case Study; Cells; Characteristics; Clinical; Clinical Trials; Clonal Evolution; DNA Sequence Alteration; Development; Diagnostic; Disease; Disease Resistance; Engraftment; Epidermal Growth Factor Receptor; Equilibrium; Erlotinib; Evaluation; Event; Evolution; Excision; Genetic; Genomics; Glioblastoma; Glioma; Goals; Grant; Growth; Health; Heterogeneity; High-Throughput Nucleotide Sequencing; Individual; Knowledge; Malignant Neoplasms; Malignant neoplasm of lung; Measures; Methodology; Methods; Microscopic; Mus; Mutate; Nature; Newly Diagnosed; Oncology; Operative Surgical Procedures; Outcome; Pathway interactions; Patients; Pattern; Phylogenetic Analysis; Play; Population; Process; Property; Radiation; Radiation therapy; Recurrence; Recurrent disease; Relapse; Residual state; Resistance; Role; Sampling; Solid Neoplasm; Stress; Structure; System; Techniques; The Cancer Genome Atlas; Therapeutic; Therapeutic Uses; Toxic effect; Transplantation; Treatment Protocols; Trees; Tumor Biology; Tumor Debulking; Tumor-Derived; United States; Xenograft Model; Xenograft procedure; base; cancer genome; chemotherapy; fluidity; genomic data; genomic profiles; improved; improved outcome; mathematical methods; melanoma; mutant; neoplastic cell; novel; novel diagnostics; novel therapeutics; regional difference; small hairpin RNA; standard care; standard of care; targeted agent; targeted treatment; temozolomide; therapy resistant; treatment response; tumor; tumor progression

DESCRIPTION (provided by applicant): Glioblastomas (GBM) are the most common and aggressive type of adult brain tumors. Despite the standard of care, concomitant radiation and temozolomide based chemotherapy treatment, disease relapse typically occurs within a year for most patients. The complexity of the disease is underlined by recent discoveries of regional differences within the tumor that may contribute to therapy resistance. Through high throughput sequencing and tagging of individual GBM cells using shRNA barcodes, the heterogeneity of the tumor cell mix can be investigated under variable circumstances. The first goal of this grant is to apply this methodology when growing GBMs in mice, to evaluate the degree of complexity after proliferation in absence of therapeutic challenges. To study whether the heterogeneity of the tumor cell mix contributes to the sensitivity to chemo- and radio-therapy, we will apply standard treatment protocols to mouse xenografts and analyze the cellular diversity of the resulting tumors. The second aim of this grant is to evaluate whether tumor complexity can be modulated using therapeutics and whether this property plays a role in developing treatment resistance. Through computational and mathematical approaches, the genomic abnormality profile can be analyzed to infer clonal and subclonal cell populations. When applied to multiple related genomic profiles, such as from diagnostic tumors and matching post-treatment tumor biopsies, patterns of clonal evolution can be uncovered. These can be related to patient features such as outcome, but also to tumor biology characteristics such as the presence of specific genomic alterations. The final aim of this grant is to construct the evolutionary path tha GBM take to escape treatment and result in recurrence. In summary, by evaluating the patterns of clonal evolution of single cells under normal growth properties, under the stress of treatment and in patient tumors, this proposal aims to improve our understanding of why GBM are so resistant to the toxic effects of therapy.

描述（由申请人提供）：胶质母细胞瘤（GBM）是最常见且最具侵袭性的成人脑肿瘤类型。尽管采用标准护理、伴随放疗和基于替莫唑胺的化疗治疗，但大多数患者通常会在一年内出现疾病复发。最近发现肿瘤内的区域差异可能导致治疗抵抗，这突显了该疾病的复杂性。通过使用 shRNA 条形码对单个 GBM 细胞进行高通量测序和标记，可以在不同情况下研究肿瘤细胞混合物的异质性。这笔资助的第一个目标是在小鼠体内培养 GBM 时应用这种方法，以评估在没有治疗挑战的情况下增殖后的复杂程度。为了研究肿瘤细胞混合物的异质性是否会影响对化疗和放疗的敏感性，我们将对小鼠异种移植物应用标准治疗方案并分析所得肿瘤的细胞多样性。这笔资助的第二个目的是评估肿瘤复杂性是否可以通过治疗来调节，以及这种特性是否在产生治疗耐药性中发挥作用。通过计算和数学方法，可以分析基因组异常概况以推断克隆和亚克隆细胞群。当应用于多个相关的基因组图谱（例如来自诊断肿瘤和匹配的治疗后肿瘤活检）时，可以揭示克隆进化的模式。这些可能与患者特征（例如结果）有关，也与肿瘤生物学特征（例如特定基因组改变的存在）有关。这笔赠款的最终目的是构建 GBM 逃避治疗并导致复发的进化路径。总之，通过评估单细胞在正常生长特性下、治疗压力下和患者肿瘤中的克隆进化模式，该提案旨在提高我们对为什么 GBM 对治疗的毒性作用具有如此抵抗力的理解。