Utilization of Peripheral Blood Exosomes to Detect Genetic and Protein Markers in Pediatric Solid Tumor Patients

利用外周血外泌体检测儿科实体瘤患者的遗传和蛋白质标志物

• 批准号: 10226046
• 负责人: AMY Leanne WALZ
• 金额: $ 12.04万
• 依托单位: LURIE CHILDREN'S HOSPITAL OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10226046
• 关键词: Alveolar Rhabdomyosarcoma; Antibodies; Biological Markers; Biopsy; Blood; Blood specimen; CD81 gene; Cell Line; Cells; Characteristics; Child; Childhood Solid Neoplasm; Clinical; Clinical Trials Cooperative Group; Column Chromatography; Combined Modality Therapy; DNA; Desmoplastic Small Round Cell Tumor; Detection; Diagnosis; EWS-FLI1 fusion protein; Early Diagnosis; Ensure; Enzyme-Linked Immunosorbent Assay; Evaluation; Ewings sarcoma; Ewings sarcoma-primitive neuroectodermal tumor; Exposure to; Future; Genetic Markers; Genomics; Goals; Heterogeneity; Image; Knowledge; Laboratories; Membrane; Messenger RNA; Methods; MicroRNAs; Molecular Sieve Chromatography; Monitor; Mutation; NCAM1 gene; Neoplasm Circulating Cells; Newly Diagnosed; Patients; Pediatric Neoplasm; Pediatric Oncology Group; Plasma; Preparation; Primary Neoplasm; Procedures; Proteins; RNA; RNA Degradation; Radiation; Recurrence; Reproducibility; Routine Diagnostic Tests; Sampling; Scanning; Sedation procedure; Sensitivity and Specificity; Solid Neoplasm; Surface; Technology; Testing; Tissues; Transcript; Tumor-Derived; Validation; exosome; neoplastic cell; pediatric patients; peripheral blood; protein biomarkers; research clinical testing; response; serial imaging; synovial sarcoma; treatment response; tumor; vesicular release

PROJECT SUMMARY The diagnosis of a pediatric solid tumor currently requires a tissue biopsy, which is an invasive procedure that is not able to capture the heterogeneity present within the entire tumor. These children then typically receive a combination of therapies, with response determined by serial imaging. This approach is complicated by exposure to radiation during imaging and requirements for sedation in younger children. Therefore, superior methods are needed for diagnosis and for monitoring of response to treatment in children with solid tumors. Recently, several technologies have been developed to analyze blood samples for the genomic presence of solid tumors within circulating free DNA or circulating tumor cells. However, their clinical utility is limited in pediatric neoplasms, which often rely on the detection of fusion transcripts for diagnosis. Exosomes, which are small, membrane-enclosed vesicles released from cells, contain mRNA, miRNA, and protein cargo unique to their cell of origin. Importantly, exosomes protect RNA from degradation and could provide the ideal biomarker for diagnosis, monitoring treatment response, and detection of early recurrence in children with solid tumors. However, further evaluation of the methods utilized for exosome isolation and cargo evaluation are needed to ensure that detection is reliable and reproducible prior to moving forward with the use of exosomes as biomarkers in the clinical realm. The proposed studies will identify the appropriate pre-analytic conditions for the reliable isolation of exosomes from the blood of pediatric patients with solid tumors containing known fusion transcripts identified through routine diagnostic testing, including Ewing Sarcoma/PNET, synovial sarcoma, alveolar rhabdomyosarcoma, and desmoplastic small round cell tumor, as well as for the reproducible detection of tumor-specific markers within the isolated exosomes. We will isolate exosomes from peripheral blood samples from a total of 50 healthy children and 30 children with newly diagnosed solid tumors during this study, and will apply clinical laboratory validation concepts of accuracy, precision, and analytic sensitivity and specificity to establish the reliability and utility of exosomal cargo as biomarkers in children with solid tumors. The overall goal of this study is to provide sufficient information to allow for the proposal of studies within the Children’s Oncology Group cooperative clinical trials to investigate the utility of exosome isolation and cargo analysis. We anticipate that within 10 years, the analysis of peripheral blood for exosome characteristics may provide a non-invasive method for establishing the diagnosis, monitoring treatment response, and detecting early recurrence in children with solid tumors.

项目概要 目前，儿科实体瘤的诊断需要组织活检，这是一种侵入性的方法 该程序无法捕获整个肿瘤内存在的异质性。 然后，儿童通常会接受多种疗法的组合，并根据系列结果确定反应 这种方法由于在成像过程中暴露于辐射而变得复杂。 年幼儿童的镇静要求因此需要更先进的方法。 最近，用于实体瘤儿童的诊断和治疗反应监测。 已经开发出多种技术来分析血液样本中的基因组存在 循环游离 DNA 或循环肿瘤细胞内的实体瘤然而，它们的临床用途。 仅限于儿科肿瘤，这通常依赖于融合转录本的检测 外泌体是从细胞中释放出来的小型膜封闭囊泡， 含有其来源细胞特有的 mRNA、miRNA 和蛋白质货物，重要的是，外泌体。 保护 RNA 免遭降解，并可为诊断、监测提供理想的生物标志物 然而，实体瘤儿童的治疗反应和早期复发的检测。 进一步评估用于外泌体分离和货物评估的方法 在继续使用之前需要确保检测可靠且可重复 拟议的研究将确定外泌体作为临床领域的生物标志物。 从血液中可靠分离外泌体的适当的分析前条件 患有实体瘤的儿科患者含有通过常规方法鉴定出的已知融合转录本 诊断测试，包括尤文肉瘤/PNET、滑膜肉瘤、肺泡肉瘤 横纹肌肉瘤和促结缔组织增生性小圆细胞肿瘤，以及可重复的 检测分离的外泌体中的肿瘤特异性标记物我们将分离外泌体。 采集自 50 名健康儿童和 30 名新发儿童的外周血样本 在本研究期间诊断出实体瘤，并将应用以下临床实验室验证概念 准确度、精密度、分析灵敏度和特异性，以建立可靠性和实用性 外泌体货物作为实体瘤儿童的生物标志物。 提供足够的信息以允许儿童肿瘤学内的研究提案 小组合作临床试验研究外泌体分离和货物的效用 我们预计在 10 年内，外周血中的外泌体分析将得到实现。 特征可以提供一种非侵入性方法来建立诊断、监测 治疗反应，并检测患有实体瘤的儿童的早期复发。