Circulating Cell-Free DNA as a Personalized Biomarker to Diagnose and Monitor Glioblastoma

循环游离 DNA 作为诊断和监测胶质母细胞瘤的个性化生物标志物

• 批准号: 10078947
• 负责人: Hunter Reeve Underhill
• 金额: $ 34.88万
• 依托单位: UNIVERSITY OF UTAH
• 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 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10078947
• 关键词: Adult; Affect; Animal Model; Apoptosis; Appearance; Attenuated; Biological Markers; Blood Circulation; Brain; Brain Neoplasms; Catalogs; Cell Fraction; Cells; Clinical; Clinical Management; Custom; DNA; Data; Detection; Diagnosis; Diagnostic; Diagnostic Procedure; Disease; Excision; Frequencies; Gene Frequency; Genetic Heterogeneity; Genetic Variation; Genotype; Glioblastoma; Glioma; Goals; Histologic; Human; Image; Knowledge; Length; Lesion; Malignant - descriptor; Malignant Neoplasms; Methods; Modeling; Molecular Profiling; Monitor; Neoplasm Metastasis; Neuraxis; Neurologic Deficit; Newly Diagnosed; Noise; Operative Surgical Procedures; Patient Care; Patients; Perfusion; Plasma; Plasma Cells; Point Mutation; Positioning Attribute; Primary Brain Neoplasms; Procedures; Radiation necrosis; Rattus; Recurrence; Recurrent disease; Risk; Sampling; Solid Neoplasm; Source; Specificity; Stable Disease; Stroke; Testing; Time; Tissue Sample; Translating; Translations; Tumor Tissue; Tumor-Derived; Variant; Work; Xenograft procedure; base; cell free DNA; design; diagnostic accuracy; genetic variant; improved; next generation sequencing; profiles in patients; promoter; response; stem-like cell; success; treatment effect; tumor; tumor DNA; variant detection

PROJECT SUMMARY/ABSTRACT Glioblastoma (GBM) is the most common and most aggressive adult primary brain tumor. Regardless of therapy, the median survival time is less than 15 months as GBM nearly always recurs. Diagnosing GBM currently depends on acquiring tumor tissue for histologic examination to differentiate GBM from other types of brain lesions with a similar imaging appearance. Detecting GBM recurrence is also challenging as treatment effects such as pseudoprogression and radiation necrosis mimic recurrent disease. Thus, risk-associated diagnostic invasive procedures remain a critical aspect of GBM patient care even though surgically acquired neurologic deficits may reduce an already short survival time. Therefore, a non-invasive approach to diagnose GBM and identify true recurrence has the potential to directly impact patient care throughout the entire disease course. DNA released from cells during programmed cell death, termed cell-free DNA, is an emerging biomarker for diagnosing human cancers and monitoring response to therapy. Detection of tumor-derived cell- free DNA in plasma, also known as circulating tumor DNA (ctDNA), has previously been largely unsuccessful in humans with GBM due to an inability to find the ctDNA amongst the abundant background of normally occurring cell-free DNA. The large intra- and inter-tumor genetic heterogeneity of GBM has hindered searches for specific tumor variants in cell-free DNA, while the rarity of GBM to metastasize beyond the central nervous system has considerably restricted the quantity of ctDNA present. However, our uncovering of ctDNA in a xenograft brain tumor model using human GBM stem-like cells strongly supports the feasibility of detecting GBM-derived ctDNA in humans. Moreover, our previous success in the animal model provides direction for the human translation – reducing or eliminating noise associated with next-generation sequencing (NGS) that interferes with the detection of very low frequency variants is necessary to allow searches for ctDNA that are not dependent on a priori knowledge of solid tumor variants. In this proposal, we attenuate NGS-related noise and demonstrate the unbiased detection of GBM-derived ctDNA at time of initial diagnosis and at recurrence. We also show that identification of GBM solid tumor DNA variants in ctDNA is enhanced by reducing errors associated with NGS to improve sampling of the solid tumor genetic heterogeneity typical of GBM. Therefore, this proposal seeks to translate plasma cell-free DNA as a biomarker to detect GBM in humans by suppressing errors associated with NGS to improve variant detection at very low allele frequencies. The successful application of ctDNA biomarkers to non-invasively assess GBM will directly affect patient care by enabling the optimization of clinical management prior to or instead of risk-associated invasive diagnostic procedures.

项目摘要/摘要 胶质母细胞瘤（GBM）是最常见，最具侵略性的成年原发性脑肿瘤。不管 治疗，中位生存时间少于15个月，因为GBM几乎总是复发。诊断GBM 目前取决于获得组织学检查的肿瘤组织，以区分GBM与其他类型的 具有相似成像外观的脑病变。检测GBM复发也很具有挑战性 诸如伪雌性和辐射坏死模仿复发性疾病之类的作用。那是与风险相关的 诊断性侵入性程序仍然是GBM患者护理的关键方面，即使手术已获得 神经系统防御可能会减少已经很短的生存时间。因此，一种非侵入性诊断方法 GBM并确定真正的复发有可能直接影响整个疾病的患者护理 课程。在编程细胞死亡期间从细胞释放的DNA称为无细胞DNA，是出现的 用于诊断人类癌症的生物标志物并监测对治疗的反应。检测肿瘤衍生的细胞 血浆中的游离DNA，也称为循环肿瘤DNA（CTDNA），以前已经很大程度上失败了 在具有GBM的人类中，由于无法在正常的绝对背景中找到ctDNA 发生无细胞的DNA。 GBM的大型肿瘤内和肿瘤间遗传异质性阻碍了搜索 对于无细胞DNA中的特定肿瘤变异，而GBM的稀有性转移到了中枢神经系统之外 系统仔细限制了存在的ctDNA的数量。但是，我们在a中发现ctDNA 使用人类GBM干细胞的色谱脑肿瘤模型强烈支持检测的可行性 GBM衍生的ctDNA在人类中。而且，我们以前在动物模型中的成功为 人翻译 - 减少或消除与下一代测序（NGS）相关的噪声（NGS） 需要干扰检测非常低的频率变体以允许搜索ctDNA的搜索 不取决于实体瘤变体的先验知识。在此提案中，我们减弱了与NGS相关的噪声 并在初始诊断时和复发时证明了GBM衍生的ctDNA的无偏检测。 我们还表明，通过减少误差，可以增强CTDNA中GBM实体瘤DNA变异的鉴定 与NGS相关，以改善GBM典型的实体瘤遗传异质性的采样。所以， 该建议旨在将无等离子可数的DNA翻译为一种生物标志物，以通过抑制人类检测GBM 与NG相关的误差以改善非常低等位基因频率的变异检测。成功 应用CTDNA生物标志物在非侵入性评估GBM上，将直接影响患者护理 在临床管理之前或而不是与风险相关的侵入性诊断程序进行优化。