Harnessing artificial microRNA clusters against glioblastoma epigenetic plasticity and resistance to therapy

利用人工 microRNA 簇对抗胶质母细胞瘤表观遗传可塑性和治疗耐药性

• 批准号: 10376238
• 负责人: Pier Paolo Peruzzi
• 金额: $ 41.52万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10376238
• 关键词: Abate; Address; Adult; Animal Model; BMI1 gene; Biogenesis; Biological; Biology; Brain; Caring; Cells; Characteristics; Chromatin; Clinical; Complex; DNA Damage; Data; Defense Mechanisms; Diagnosis; Diffusion; Disease; EZH2 gene; Engineering; Epigenetic Process; Genetic Structures; Genetic Transcription; Glioblastoma; Goals; Hybrids; Immune; Immunotherapy; In Vitro; Individual; KDM1A gene; Length; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of brain; Mediating; Messenger RNA; Methods; MicroRNAs; Nature; Neuronal Differentiation; Nucleotides; Oncogenic; Outcome; PRC1 Protein; Pathway interactions; Patients; Pharmacology; Primary Brain Neoplasms; Proteins; RNA; Radiation; Regulation; Research; Resistance; Role; Stress; Synthetic Genes; Testing; Therapeutic; Time; Toxic effect; Transcript; Transgenes; Tumor Biology; Untranslated RNA; Work; cellular targeting; combinatorial; design; design and construction; experimental study; extracellular vesicles; gain of function; gene therapy; genotoxicity; histone demethylase; immunoregulation; improved; in vivo; innovation; loss of function; neoplastic cell; novel; novel strategies; prevent; response; scaffold; sound; synergism; temozolomide; therapeutic miRNA; therapeutic target; therapy resistant; trafficking; translational approach; tumor; tumor microenvironment

Glioblastoma is the most common primary brain cancer in adults and remains a deadly disease. Despite intense research and clinical efforts, its survival has not significantly improved for the past several decades. A major reason for this recalcitrant nature is the tumor’s ability to escape toxicity from currently available therapies, including Temozolomide (TMZ), radiation, and immune-modulation. This resistance depends on the activation of defense mechanisms in response to damage, and is mediated epigenetically by a complex, multi- component machinery that governs the transcriptional status of the tumor cells. Our studies have defined oncogenic proteins EZH2, BMI1 and LSD1 as crucial components of this epigenetic response. We have also demonstrated that they are regulated by specific microRNAs (miR-124, miR-128 and miR-137), co-expressed in normal brain, and simultaneously lost in glioblastoma. The re-expression of these three microRNAs in glioblastoma provides a strong inhibition against the three proteins and results in a very relevant sensitization of tumor cells to stress, both in vitro and in vivo. The overall goal of this proposal is two-fold: first, to characterize the mechanism whereby the co-activation of EZH2, BMI1 and LSD1 mediates tumor survival, establishing its necessary role in this response; Second, to develop a microRNA-mediated strategy to abate this epigenetic shield that is vital to the tumor, to support a gene therapy approach synergistic with current standard therapies. AIM1 proposes experiments to dissect the role of the EZH2/BMI1/LSD1 epigenetic complex in tumor responses against Temozolomide, radiation, and immunotherapy. AIM2 provides novel solutions to design and construct artificial genes that multiply the payload of microRNAs, and which thus function as an ideal gene therapy platform for multi-targeting this epigenetic response. AIM3 investigates the applicability of multi-microRNA-based therapy in animal models of glioblastoma, to address the vital problem of tumor resistance in vivo. INNOVATION: The proposed study to use artificial microRNA clusters for the control of the complex epigenetic tumor survival response is an innovative and heretofore unexplored approach. Also, this proposal introduces strategies to exploit the unique features of microRNAs biology, processing and intercellular trafficking in the perspective of an integrated gene therapy approach. LONG-TERM OBJECTIVE: We seek to meaningfully impact the care of glioblastoma patients through the application of principles of microRNA clustering, epigenetic interference, and synergism with other therapies.

胶质母细胞瘤是成人最常见的原发性脑癌，仍然是一种致命疾病。尽管 在过去的几十年中，激烈的研究和临床工作，其生存并没有显着改善。一个 这种顽固性的主要原因是肿瘤从目前可用的毒性中逃脱毒性的能力 疗法，包括替莫唑胺（TMZ），辐射和免疫调节。这种阻力取决于 响应损害的防御机制的激活，并通过复杂的多种多样的遗传介导 控制肿瘤细胞转录状态的组件机械。我们的研究定义了 致癌蛋白EZH2，BMI1和LSD1作为这种表观遗传反应的关键成分。我们也有 证明它们受特定的microRNA（mir-124，mir-128和mir-137）的调节，共表达 在正常的大脑中，在胶质母细胞瘤中很容易丢失。这三个microRNA在 胶质母细胞瘤对三种蛋白质提供了强烈的抑制作用，并导致非常相关的敏化 肿瘤细胞在体外和体内都有压力。该提议的总体目标是两倍：首先 表征EZH2，BMI1和LSD1的共激活的机制，介导肿瘤的存活率， 在这一反应中确立必要的作用；第二，制定microRNA介导的策略以减轻 这种对肿瘤至关重要的表观遗传屏蔽，以支持基因治疗方法与电流协同作用 标准疗法。 AIM1提案实验剖析EZH2/BMI1/LSD1表观遗传学的作用 肿瘤反应对替莫唑胺，放射线和免疫疗法的复合物。 AIM2提供了小说 设计和构建人工基因的解决方案，这些基因将microRNA的有效载荷倍增，因此 充当理想的基因治疗平台，用于多目标这种表观遗传反应。 AIM3调查了 基于多微米的疗法在胶质母细胞瘤动物模型中的适用性，以解决至关重要的问题 体内肿瘤抗性。创新：拟议的研究使用人工microRNA簇进行控制 复杂的表观遗传性肿瘤存活反应是一种创新的和迄今为止的意外方法。还， 该建议介绍了利用MicroRNA生物学，加工和 从综合基因治疗方法的角度来看，细胞间贩运。长期目标： 我们寻求通过应用原则来有意义地影响胶质母细胞瘤患者的护理 microRNA聚类，表观遗传干扰以及与其他疗法的协同作用。