Therapeutic targeting of the SWI/SNF chromatin remodeler to regulate GBM chemosensitivity

SWI/SNF 染色质重塑剂的治疗靶向调节 GBM 化学敏感性

• 批准号: 10711581
• 负责人: DUANE D MILLER
• 金额: $ 50.4万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-15 至 2028-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10711581
• 关键词: ATPase Domain; Address; Adult; Alkylating Agents; Binding; Biological; Blood - brain barrier anatomy; Blood Vessels; Brain; Bromodomain; Catalytic Domain; Cell Differentiation process; Cell Line; Cell Maintenance; Cells; Chromatin; Chromatin Modeling; Chromatin Remodeling Factor; Clinical; Complex; Coupled; DNA; Drug Design; Epigenetic Process; Foundations; Future; Gene Expression; Generations; Genes; Genetic Transcription; Glioblastoma; Goals; Histones; Human; Image; Immune; Immune response; Immunocompetent; Immunocompromised Host; In Vitro; Inflammatory; Interferons; Knock-out; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of brain; Mediating; Modeling; Molecular; Molecular Target; Mus; Necrosis; Operative Surgical Procedures; Pathway interactions; Patients; Penetrance; Pharmaceutical Preparations; Phenotype; Positioning Attribute; Primary Brain Neoplasms; Prognosis; Property; RNA Polymerase II; Reader; Recurrent tumor; Reducing Agents; Research; Resistance; Role; SMARCA4 gene; SWI/SNF Family Complex; Structure; Surface; Testing; Therapeutic; Therapeutic Intervention; Tumorigenicity; Work; analog; anti-cancer; blood-brain barrier penetration; brahma; cancer cell; candidate identification; chemotherapy; chromatin immunoprecipitation; effective therapy; genetic signature; histone modification; immunoreactivity; improved; in vivo; inhibitor; innovation; knock-down; malignant phenotype; mouse model; multidisciplinary; mutant; neoplastic cell; next generation; novel; novel therapeutic intervention; patient prognosis; pharmacologic; prevent; programs; response; small molecule; stem; stem cell self renewal; stem cells; stemness; targeted treatment; temozolomide; therapeutic target; therapy resistant; transcriptome; transcriptome sequencing; tumor; tumor microenvironment; tumorigenesis

Glioblastoma (GBM) is the most common primary malignancy of the adult brain and is among the most devastating cancers as it is invariably lethal. The perivascular niche and necrotic regions of GBM tumors is enriched for stem-like tumor cells called GBM stem cells (GSCs) that are highly resistant to therapy. Within GBM tumors there are also differentiated cells that are intrinsically resistant, or acquire resistance to, therapy. Thus, there is a critical unmet need to identify and target the molecular pathways that promote GBM cancer stemness, which will enhance GBM sensitivity to currently approved therapies. This proposal is premised on several of our key findings: 1) the BRG1 catalytic subunit of the SWI/SNF chromatin remodeling complex promotes the GBM GSC phenotype and resistance to DNA alkylating agents such as temozolomide (TMZ) used to treat GBM patients; 2) PFI-3, a small molecule BRG1 bromodomain (BRD) inhibitor (BRI), improves GSC sensitivity to DNA alkylating agents by reducing GSC stemness; 3) next generation BRIs were developed that overcome the resistance of TMZ-resistant GBM cell lines, including a potent BRG1-specific BRI (IV-255); and 4) BRIs increase expression of a subclass of interferon (IFN) response genes that are predicted to enhance the anti-tumoral host immune response. Our central hypothesis is that targeting the BRD of BRG1 selectively disrupts GBM GSCs maintenance, which enhances GBM sensitivity to approved therapies and increases GBM immunoreactivity. In Aim 1 we will restore BRG1 expression in BRG1-KD (knockdown) GSCs and in TMZ resistant GBM cells with either wild-type BRG or mutant BRG1 BRD that prevents chromatin binding. We will examine the role of the BRG1 BRD in maintaining the GBM malignant phenotype and therapeutic sensitivity in vitro and in vivo. In Aim 2 we will refine the BRI structure and develop additional small molecule BRG1 BRIs. We will define BRI activity on cancer stemness and therapeutic sensitivity on GBM in vitro and in vivo, and assess selectivity for BRD binding, drug-like properties and optimize BBB penetrance. The anticancer efficacy of BRIs on intracranial GBM tumors will be tested in both immunocompromised and immunocompetent GBM mouse models to define their impact on tumor-associated immune cells. In Aim 3 we will define how BRD targeting of BRG1 reprograms transcription to promote GBM immunoreactivity and inhibit tumorigenesis by performing global transcriptome analysis (RNA-seq), which will be coupled with BRG1 and RNA polymerase II specific ChIP. These BRD-regulated genes will define the BRG1- dependent transcriptome in GSCs, and it also will identify candidate molecular pathways that could be pharmacologically targeted with BRG1 inhibition to get cancer cell synthetic lethality. Our overarching goal is to target BRG1 therapeutically with BRIs combined with presently employed therapies as a novel and urgently required therapeutic approach for this devastating form of brain cancer.

胶质母细胞瘤（GBM）是成人大脑中最常见的主要恶性肿瘤，是最常见的原发性恶性肿瘤 毁灭性的癌症总是致命的。 GBM肿瘤的血管周期和坏死区域 富含对治疗高度抗性的称为GBM干细胞（GSC）的干肿瘤细胞。 在GBM肿瘤中，还有分化的细胞本质上具有抗性或获得抗性 到治疗。因此，识别和靶向分子途径的关键未满足需要 促进GBM癌症的干性，这将增强对当前批准疗法的GBM敏感性。这 提案以我们的几个关键发现为前提：1）SWI/SNF的BRG1催化亚基 染色质重塑络合物促进GBM GSC表型和对DNA烷基化的抗性 替莫唑胺（TMZ）等药物用于治疗GBM患者； 2）PFI-3，一个小分子BRG1 溴结构域（BRD）抑制剂（BRI）通过还原GSC来提高GSC对DNA烷基化剂的敏感性 茎； 3）开发了下一代BRI，以克服抗TMZ的GBM的抗性 细胞系，包括有效的BRG1特异性BRI（IV-255）； 4）Bris增加了子类的表达 干扰素（IFN）反应基因的预测，可增强抗肿瘤宿主免疫反应。 我们的中心假设是，针对BRG1的BRD有选择地破坏GBM GSC的维护， 这增强了GBM对批准疗法的敏感性，并提高了GBM免疫反应性。在目标1中 我们将恢复BRG1-KD（敲低）GSC中的BRG1表达以及具有TMZ抗TMZ GBM细胞 野生型BRG或突变体BRG1 BRD可防止染色质结合。我们将研究 BRG1 BRD在维持GBM恶性表型和在体外和IN的治疗敏感性方面 体内。在AIM 2中，我们将完善BRI结构并发展其他小分子BRG1 BRI。我们将 定义对癌症的BRI活性和对GBM的体外和体内GBM的治疗敏感性，以及 评估BRD结合，类似药物的特性的选择性并优化BBB渗透率。抗癌者 BRI对颅内GBM肿瘤的功效将在免疫功能低下和 免疫能力的GBM小鼠模型定义了它们对肿瘤相关免疫细胞的影响。目标 3我们将定义BRD靶向BRG1如何重新编程转录以促进GBM免疫反应性 并通过进行全局转录组分析（RNA-SEQ）抑制肿瘤发生，该分析将耦合 带有BRG1和RNA聚合酶II特异性芯片。这些受BRD调节的基因将定义BRG1- GSC中的依赖转录组，它还将确定可以是的候选分子途径 具有BRG1抑制的药理靶向以获得癌细胞合成致死性。我们的总体 目标是将BRI与BRI与目前使用的疗法结合在一起的新颖靶向BRG1。 迫切需要对这种毁灭性脑癌形式的治疗方法。

项目成果

Next-generation bromodomain inhibitors of the SWI/SNF complex enhance DNA damage and cell death in glioblastoma.

• DOI: 10.1111/jcmm.17907
• 发表时间: 2023-09
• 期刊: JOURNAL OF CELLULAR AND MOLECULAR MEDICINE
• 影响因子: 5.3
• 作者: Yang, Chuanhe;He, Yali;Wang, Yinan;McKinnon, Peter J.;Shahani, Vijay;Miller, Duane D.;Pfeffer, Lawrence M.
• 通讯作者: Pfeffer, Lawrence M.