Dissecting the role of ATRX in soft tissue sarcoma development and radiation response

剖析 ATRX 在软组织肉瘤发展和放射反应中的作用

• 批准号: 10067365
• 负责人: Robert Warren Floyd
• 金额: $ 3.82万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-01 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10067365
• 关键词: ATRX gene; Affect; Biological Assay; Biological Models; Cancer Burden; Cancer Patient; Caring; Cell Line; Centromere; Clinical; Connective Tissue; Cre-LoxP; DNA Damage; DNA Repair; DNA Sequence Alteration; Data; Defect; Development; Double Strand Break Repair; Fluorescent in Situ Hybridization; Frameshift Mutation; Functional disorder; Genes; Genetic Determinism; Genetically Engineered Mouse; Genomic Instability; Glioma; Goals; Growth; Histones; Human; Immune signaling; Immunofluorescence Immunologic; Immunotherapy; Impairment; In Situ Hybridization; In Vitro; Innate Immune Response; Ionizing radiation; KRAS oncogenesis; KRASG12D; Knock-out; Knowledge; Lead; Link; Maintenance; Malignant Neoplasms; Measurement; Mental Retardation; Mission; Mitotic; Modeling; Molecular Chaperones; Mouse Cell Line; Mus; Mutate; Mutation; Natural Immunity; Nonhomologous DNA End Joining; Oncogenes; Pathway interactions; Patients; Prediction of Response to Therapy; Prevalence; Prognosis; Proteins; Public Health; Radiation; Radiation Tolerance; Radiation therapy; Radiosensitization; Repetitive Sequence; Research; Research Personnel; Role; Sampling; Signal Transduction; Soft tissue sarcoma; Stains; Stimulator of Interferon Genes; Survival Rate; System; Technology; Telomerase; Testing; The Cancer Genome Atlas; Therapeutic Intervention; Tumor Cell Line; Western Blotting; alpha-Thalassemia; base; cancer type; cell growth; cell type; conditional mutant; glioma cell line; homologous recombination; immunogenicity; improved; in vivo; in vivo Model; innate immune pathways; insight; knock-down; loss of function mutation; mouse model; next generation sequencing; novel; novel therapeutics; prognostic of survival; radiation response; recombinase; response; sarcoma; segregation; telomere; treatment response; tumor; tumor behavior; tumor growth

ABSTRACT: Soft tissue sarcomas are tumors of the connective tissue that account for an estimated 12,000 new cancer cases annually and carry a poor prognosis with a five year survival rate of 50% despite treatment. An important research objective in improving therapy for soft tissue sarcoma patients is to understand how genetic mutations affect soft tissue sarcoma development and radiation response. Intriguingly, next- generation sequencing data from The Cancer Genome Atlas and other massive cancer sequencing efforts have identified Alpha Thalassemia and Mental Retardation X-linked, or ATRX, as the second most frequently mutated gene in soft tissue sarcoma. ATRX is perhaps best known for its role as regulator of alternative lengthening of telomeres (ALT), a telomerase independent tumor maintenance mechanisms found in 15% of all human cancers. Interestingly, ATRX is predictive for overall survival in multiple human cancers and researchers recently demonstrated that ATRX knockdown leads to radiosensitization in glioma cell lines. Despite the clear importance of this gene in multiple human cancers and its frequent alteration in soft tissue sarcoma, the role of ATRX in soft tissue sarcoma remains relatively unstudied. The long term goal of this project is to improve the efficacy of current therapies for soft tissue sarcoma patients and enable the development of novel therapeutics for the treatment of human cancers. The overall goal of this proposal is to determine the effect of Atrx deletion on the radiosensitivity and innate immune response of soft tissue sarcoma. To achieve this, the Cre-LoxP recombinase system has been used in genetically engineered mouse models to generate the first primary mouse model of soft tissue sarcoma with ATRX deletion. The central hypothesis is that loss of ATRX impairs DNA damage repair, delays tumor development and increases radiosensitivity in soft tissue sarcoma. To test this hypothesis, primary soft tissue sarcomas with ATRX deletion will be compared to primary soft tissue sarcomas that retain Atrx. Using in vitro and in vivo model systems generated using Cre-LoxP and dual recombinase technologies, I will test this hypothesis in three specific aims: Aim 1: Investigate the effect of Atrx deletion on tumor growth and repetitive element associated mitotic dysfunction in a primary mouse model of soft tissue sarcoma Aim 2: Determine the role of ATRX in DNA damage repair and sarcoma response to ionizing radiation Aim 3: Determine the effect of Atrx deletion on radiation induced mitotic dysfunction and cGAS-STING innate immune signaling in sarcoma

摘要： 软组织肉瘤是结缔组织肿瘤，估计每年有12,000例新的 每年都有癌症病例，尽管接受了治疗，但预后很差，五年生存率为50%。 改善软组织肉瘤患者治疗的一个重要研究目标是了解 基因突变如何影响软组织肉瘤的发展和辐射反应。有趣的是，接下来- 来自癌症基因组图谱和其他大规模癌症测序的一代测序数据 努力确定阿尔法地中海贫血和智力迟钝X连锁，或ATRX，作为第二大 在软组织肉瘤中频繁突变的基因。ATRX最为人所知的可能是其作为监管机构的作用， 端粒替代延长（ALT），一种不依赖于端粒的肿瘤维持机制 在15%的人类癌症中发现。有趣的是，ATRX可预测多个患者的总生存率。 人类癌症和研究人员最近证明，ATRX敲低导致放射增敏， 在神经胶质瘤细胞系中。尽管该基因在多种人类癌症中的重要性显而易见，而且其频繁的 尽管ATRX在软组织肉瘤中的作用与其在软组织肉瘤中的作用不同，但ATRX在软组织肉瘤中的作用仍然相对未被研究。 本项目的长期目标是提高目前软组织肉瘤治疗的疗效 并且能够开发用于治疗人类癌症的新疗法。整体 该建议的目的是确定Atrx缺失对放射敏感性和先天免疫的影响。 软组织肉瘤的反应。为了实现这一点，Cre-LoxP重组酶系统已被用于 基因工程小鼠模型，以产生第一个软组织肉瘤的原代小鼠模型， ATRX删除。核心假设是ATRX的缺失损害DNA损伤修复，延迟肿瘤生长， 发展和增加软组织肉瘤的放射敏感性。为了验证这一假设， 将ATRX缺失的组织肉瘤与保留Atrx的原发性软组织肉瘤进行比较。 使用Cre-LoxP和双重组酶技术产生的体外和体内模型系统， 将在三个具体目标中检验这一假设： 目的1：研究Atrx基因缺失对肿瘤生长及相关重复序列的影响。 软组织肉瘤小鼠模型中有丝分裂功能障碍 目的2：确定ATRX在DNA损伤修复和肉瘤对电离辐射反应中的作用 目的3：确定Atrx缺失对辐射诱导的有丝分裂功能障碍和cGAS-STING的影响 肉瘤中的天然免疫信号