The Prognostic Significance and Mechanistic Determination of Chromatin Remodeling Biomarkers in Non-Functional Pancreatic Neuroendocrine Tumor

非功能性胰腺神经内分泌肿瘤染色质重塑生物标志物的预后意义和机制确定

• 批准号: 10451759
• 负责人: Aatur Dilip Singhi
• 金额: $ 59.71万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-15 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10451759
• 关键词: ARID1A gene; ATRX gene; Abdomen; Address; Affect; Autopsy; Behavior; Biological Assay; Biological Markers; Cells; Cessation of life; Chromatin; Chromatin Structure; Clinical; Common Neoplasm; DAXX gene; DNA Sequence Alteration; Detection; Development; Disease; Disease-Free Survival; Distant Metastasis; Epigenetic Process; Excision; Gene Expression Regulation; Genes; Genetic Transcription; Genomics; Goals; High Prevalence; Histone H3; Histones; Incidence; Individual; Indolent; International; Islet Cell Tumor; Lead; Maintenance; Metastatic to; Methods; Molecular; Molecular Chaperones; Mutation; Neoplasm Metastasis; Neoplasms; Nonmetastatic; Operative Surgical Procedures; Pathogenesis; Pathway interactions; Patients; Pattern; Play; Postoperative Period; Prognostic Marker; Prospective Studies; Proteins; Proteomics; Recurrence; Regulation; Regulator Genes; Reporting; Resolution; Retrospective Studies; Role; Sampling; Specimen; Staging System; Stratification; System; Telomerase; Testing; United States; Validation; cancer cell; chromatin remodeling; clinical implementation; clinically significant; cohort; epigenomics; imaging modality; improved; innovation; insight; nanoscale; neoplastic cell; novel; overtreatment; pancreatic neoplasm; patient biomarkers; patient prognosis; patient stratification; patient subsets; prognostic; prognostic assays; prognostic performance; prognostic significance; prospective; radiological imaging; reconstitution; research clinical testing; response; telomere; transcriptome sequencing; tumor; tumor progression; validation studies

Project Summary/Abstract Non-functional pancreatic neuroendocrine tumors (NF-PanNETs) are a heterogeneous group of neoplasms with increasing incidence and ill-defined pathobiology. While many NF-PanNETs are indolent and remain stable for years, a significant subset may behave aggressively and metastasize widely. Thus, the increasing and frequent detection of NF-PanNETs presents a treatment dilemma. Current prognostic parameters and systems are susceptible to interpretation errors, sampling issues, and do not accurately reflect the clinical behavior of these neoplasms. Hence, additional biomarkers are needed to improve the prognostic stratification of patients with NF-PanNETs. Previously, we and others have reported recurrent genomic alterations in ATRX and DAXX are associated with metastatic progression of NF-PanNETs. Mutations in these genes result in loss of their respective proteins and coincide with the alternative lengthening of telomeres (ALT), a telomerase-independent maintenance mechanism. We have also shown that loss of ATRX/DAXX and ALT correlate with the development of metastases, are prognostic biomarkers for shorter disease-free survival and are independent of other prognostic clinicopathologic parameters. Thus, ATRX/DAXX and ALT represent promising biomarkers. However, they have not been evaluated prospectively nor in preoperative specimens, where prognostic stratification is important for patient management. In addition, only 50% of metastatic NF-PanNETs harbor inactivation of ATRX/DAXX and the presence of ALT. To identify additional biomarkers, we recently profiled ATRX/DAXX wild type metastatic NF-PanNETs and reported recurrent alterations in SETD2/H3K36me3 and ARID1A, which similar to ATRX and DAXX are chromatin regulating genes. We therefore hypothesize that the metastatic progression of NF-PanNETs is characterized by changes in chromatin regulation and determining the key genomic and epigenomic hallmarks will not only improve the prognostic stratification of patients with NF-PanNETs, but advance our understanding of the pathogenesis of this increasingly prevalent disease. Aim 1 will be to develop and clinically validate preoperative prognostic biomarker assays for NF-PanNETs. Through both retrospective and prospective studies, we will determine the prognostic performance and significance of ATRX, DAXX, ALT, H3K36me3 and ARID1A. For Aim 2, we plan to define the chromatin patterns at different epigenetic states in the metastatic progression of NF-PanNETs. Considering ATRX, DAXX, SETD2 and ARID1A are chromatin regulators, we will evaluate the nanoscale chromatin structure and affected molecular pathways of various NF-PanNET states using PathSTORM and CUT&RUN/RNA-seq assays. Finally, Aim 3 will investigate the adaptive response of ATRX/DAXX inactivation and ALT initiation. We have discovered that the histone chaperone, HIRA, can reconstitute telomeric chromatin and function of ATRX/DAXX deficiency ALT cancer cells. Within this aim, we will delineate the role of HIRA in regulating telomeric integrity, chromatin and transcription. Overall, this proposal will lead to the development of a clinically available assay for patient management, insight in the pathognomonic chromatin structural changes and identify novel drivers/pathways for not only NF-PanNETs, but likely other ALT-associated neoplasms.

项目概要/摘要 非功能性胰腺神经内分泌肿瘤（NF-PanNET）是一组异质性肿瘤，其发病率不断增加 发病率和不明确的病理学。虽然许多 NF-PanNET 很懒惰并且多年来保持稳定，但一个重要的 亚群可能表现出攻击性并广泛转移。因此，NF-PanNET 的检测越来越频繁 呈现出治疗困境。当前的预后参数和系统容易受到解释错误、采样的影响 问题，并且不能准确反映这些肿瘤的临床行为。因此，需要额外的生物标志物 改善 NF-PanNET 患者的预后分层。此前，我们和其他人曾报告过复发性 ATRX 和 DAXX 的基因组改变与 NF-PanNET 的转移进展相关。这些突变 基因导致各自蛋白质的丢失，并与端粒（ALT）的选择性延长相一致，这是一种 不依赖端粒酶的维持机制。我们还表明 ATRX/DAXX 和 ALT 的丢失与 转移的发展，是较短无病生存期的预后生物标志物，并且独立于其他因素 预后临床病理参数。因此，ATRX/DAXX 和 ALT 代表了有前途的生物标志物。然而，他们 尚未对术前标本进行前瞻性评估，其中预后分层对患者很重要 管理。此外，只有 50% 的转移性 NF-PanNET 具有 ATRX/DAXX 失活以及存在 ALT。为了识别其他生物标志物，我们最近分析了 ATRX/DAXX 野生型转移性 NF-PanNET 并报告 SETD2/H3K36me3 和 ARID1A 的反复改变，与 ATRX 和 DAXX 类似，都是染色质调节基因。 因此，我们假设 NF-PanNET 的转移进展以染色质的变化为特征 调控和确定关键基因组和表观基因组标志不仅会改善预后分层 NF-PanNET 患者的研究，但增进了我们对这种日益流行疾病发病机制的理解。 目标 1 是开发 NF-PanNET 的术前预后生物标志物测定并进行临床验证。通过两者 回顾性和前瞻性研究，我们将确定 ATRX、DAXX、 ALT、H3K36me3 和 ARID1A。对于目标 2，我们计划定义不同表观遗传状态下的染色质模式 NF-PanNET 的转移进展。考虑到 ATRX、DAXX、SETD2 和 ARID1A 是染色质调节因子，我们将 使用 NF-PanNET 状态评估纳米级染色质结构和受影响的分子途径 PathSTORM 和 CUT&RUN/RNA-seq 检测。最后，Aim 3 将研究 ATRX/DAXX 的自适应响应 失活和 ALT 启动。我们发现组蛋白伴侣 HIRA 可以重建端粒染色质 以及 ATRX/DAXX 缺陷 ALT 癌细胞的功能。在此目标范围内，我们将描述 HIRA 在监管方面的作用 端粒完整性、染色质和转录。总体而言，该提案将导致临床可用的药物的开发 用于患者管理的分析，洞察特征性染色质结构变化并识别新的 不仅是 NF-PanNET，而且可能是其他 ALT 相关肿瘤的驱动因素/途径。