Understanding how ZFHX4 drives the glioblastoma tumor propagating cell state

了解 ZFHX4 如何驱动胶质母细胞瘤肿瘤增殖细胞状态

• 批准号: 10455744
• 负责人: Milan Girish Chheda
• 金额: $ 34.45万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10455744
• 关键词: Address; Binding Sites; Biochemical; Biochemical Genetics; Biological; Biological Assay; Brain Neoplasms; CHD4 gene; Cell Line; Cell Survival; Cell physiology; Cells; Cessation of life; ChIP-seq; Coiled-Coil Domain; Colon Carcinoma; Complex; Confocal Microscopy; DNA Damage; DNA Repair; Data; Deacetylase; Deterioration; Etoposide; Funding; Genes; Genetic; Genetic Transcription; Genome; Glioblastoma; Goals; Growth; Homeobox; Human; In Vitro; Knowledge; Lasers; Lead; Maintenance; Malignant neoplasm of prostate; Mediating; Methods; Mus; Neurologic; Nucleic Acid Regulatory Sequences; Nucleosomes; Oncogenic; Patients; Persons; Play; Population; Primary Brain Neoplasms; Prognosis; Property; Proteins; Public Health; Recurrence; Research; Resistance; Resolution; Role; SMARCA3 gene; Site; Structure; Techniques; Technology; Testing; Transcriptional Regulation; Work; Xenograft Model; Zinc Fingers; angiogenesis; base; cancer stem cell; epigenetic regulation; experimental study; homologous recombination; in vivo; innovation; insight; leukemia; malignant breast neoplasm; member; migration; nervous system disorder; new technology; recruit; repaired; response; self-renewal; stem cell biomarkers; stem cells; temozolomide; therapy resistant; transcription factor; tumor; tumor xenograft

Project Summary Glioblastoma (GBM) is a primary brain tumor that causes significant neurological deterioration and death. Maximally treated glioblastoma (GBM) recurs in 6 months on average, and it kills most people in less than 2 years. A major reason for inevitable recurrence may be the existence of tumor propagating cells (TPCs), sometimes known as cancer stem cells. No current therapy specifically targets or eradicates TPCs, and our mechanistic understanding of these highly resistant cells is limited. We discovered a transcription factor, ZFHX4, that is required for the TPC state, and its suppression significantly prolongs survival in a mouse xenograft model. The higher the expression of ZFHX4 in patient tumors, the shorter they live. The overall objective of our proposal is to determine why ZFHX4 is required for the oncogenic, TPC state and to determine which aspects of GBM function it controls. We found that ZFHX4 interacts with chromodomain-helicase-DNA- binding protein 4 (CHD4), a core member of the nucleosome remodeling and deacetylase (NuRD) complex. Moreover, we found that, like ZFHX4, CHD4 is required to maintain the TPC state. We also recently discovered that ZFHX4 interacts with SMARCA3 and PARP, which are both required for efficient DNA repair, a hallmark of TPCs. We hypothesize that ZFHX4 transcriptionally drives expression of key GBM genes involved in self-renewal, growth, and migration, and that through interactions with distinct proteins, it also plays a role in DNA damage response. In Aim 1, we will determine the essential function and targets of ZFHX4 in GBM. In Aim 2, we will evaluate how ZFHX4 interacts with CHD4, and determine the biological consequences of their interaction. In Aim 3, we will determine why ZFHX4 is required for TPC survival after DNA damage. We will use biochemical, genetic, and cell biological approaches, including innovative new technologies like calling cards to track ZFHX4 binding sites under specific conditions, super-resolution confocal microscopy to study ZFHX4 localization and inform structure-function analyses, and DNA damaging laser microbeams to study the role of ZFHX4 in DNA damage. Successful execution of these aims will provide important insight into why ZFHX4 is required for the TPC state and fundamental knowledge about this transcription factor and epigenetic regulation in glioblastoma. These studies will help clarify why therapy resistant TPCs cause a significant burden of neurological disease. Our findings will have a significant impact because they will eventually lead to treatments that inhibit ZFHX4 function, reduce the TPC burden in patients, and extend survival.

项目概要 胶质母细胞瘤（GBM）是一种原发性脑肿瘤，可导致严重的神经功能恶化和死亡。 接受最大限度治疗的胶质母细胞瘤 (GBM) 平均会在 6 个月内复发，并且会在不到 2 个月的时间内杀死大多数人 年。不可避免的复发的一个主要原因可能是肿瘤增殖细胞（TPC）的存在， 有时被称为癌症干细胞。目前没有专门针对或根除 TPC 的治疗方法，我们的 对这些高耐药细胞的机制了解有限。我们发现了一个转录因子 ZFHX4，TPC 状态所必需的，其抑制可显着延长小鼠的生存期 异种移植模型。患者肿瘤中 ZFHX4 的表达越高，他们的寿命就越短。整体 我们提案的目标是确定为什么 ZFHX4 是致癌、TPC 状态所必需的，并确定 它控制 GBM 功能的哪些方面。我们发现 ZFHX4 与染色质结构域-解旋酶-DNA-相互作用 结合蛋白 4 (CHD4)，核小体重塑和脱乙酰酶 (NuRD) 复合物的核心成员。 此外，我们发现，与 ZFHX4 一样，CHD4 也需要维持 TPC 状态。我们最近也 发现 ZFHX4 与 SMARCA3 和 PARP 相互作用，这两者都是有效 DNA 修复所必需的， TPC 的标志。我们假设 ZFHX4 转录驱动相关关键 GBM 基因的表达 在自我更新、生长和迁移中，并且通过与不同蛋白质的相互作用，它还在 DNA损伤反应。在目标 1 中，我们将确定 ZFHX4 在 GBM 中的基本功能和目标。在 目标 2，我们将评估 ZFHX4 如何与 CHD4 相互作用，并确定它们的生物学后果 相互作用。在目标 3 中，我们将确定为什么 ZFHX4 对于 DNA 损伤后 TPC 的存活是必需的。我们将使用 生物化学、遗传和细胞生物学方法，包括电话卡等创新技术 跟踪特定条件下的 ZFHX4 结合位点，超分辨率共聚焦显微镜研究 ZFHX4 定位并为结构功能分析提供信息，以及 DNA 损伤激光微束以研究 DNA 损伤中的 ZFHX4。成功执行这些目标将为了解 ZFHX4 为何如此而提供重要见解 TPC 状态以及有关该转录因子和表观遗传调控的基础知识所需 在胶质母细胞瘤中。这些研究将有助于阐明为什么治疗耐药性 TPC 会造成严重的负担 神经系统疾病。我们的发现将产生重大影响，因为它们最终将导致治疗方法 抑制 ZFHX4 功能，减轻患者 TPC 负担，延长生存期。

项目成果

A Fyn romance: tumor cell Fyn kinase suppresses the immune microenvironment.

Fyn 浪漫史：肿瘤细胞 Fyn 激酶抑制免疫微环境。

• 发表时间: 2020
• 影响因子: 15.9
• 作者: Bais, Sachendra S;Chheda, Milan G
• 通讯作者: Chheda, Milan G

CHD4 regulates the DNA damage response and RAD51 expression in glioblastoma.

CHD4 调节胶质母细胞瘤中的 DNA 损伤反应和 RAD51 表达。

• 发表时间: 2019
• 影响因子: 4.6
• 作者: McKenzie, Lisa D;LeClair, John W;Miller, Kayla N;Strong, Averey D;Chan, Hilda L;Oates, Edward L;Ligon, Keith L;Brennan, Cameron W;Chheda, Milan G
• 通讯作者: Chheda, Milan G