Site-specific epigenetic activation of TP53 to improve cancer therapy

TP53 的位点特异性表观遗传激活可改善癌症治疗

• 批准号: 10258179
• 负责人: Nathaniel A. Hathaway
• 金额: $ 35万
• 依托单位: EPIGENOS BIOSCIENCE, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-16 至 2023-09-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10258179
• 关键词: ANXA5 gene; Aberrant DNA Methylation; Affect; Apoptosis; Automobile Driving; BAX gene; Binding; Biological Models; Biology; CDKN1A gene; CRISPR/Cas technology; Cancer Etiology; Cell Cycle; Cell Cycle Arrest; Cells; Cessation of life; Chemicals; Chimeric Proteins; Chromatin; Clinical; Colon Carcinoma; Colorectal Cancer; Coupled; Deacetylation; Disease; Engineering; Enzymes; Epigenetic Process; FK506; Flow Cytometry; Fluorescein; Fluorescein-5-isothiocyanate; Functional disorder; Gene Activation; Gene Expression; Generations; Genes; Genetic; Genome; Guide RNA; HCT116 Cells; Histone Acetylation; Histone Deacetylase; Histone Deacetylase Inhibitor; Histone H3; Hour; Human; In Vitro; Isothiocyanates; Lead; Malignant Neoplasms; Mediating; Methylation; Mus; Mutate; Oncogene Activation; Parents; Pathway interactions; Patient Care; Plant Roots; Polyethylene Glycols; Pre-Clinical Model; Quantitative Reverse Transcriptase PCR; Regulation; Repression; Role; SW480; Signal Pathway; Site; Stains; System; TP53 gene; Tacrolimus Binding Proteins; Technology; Therapeutic; Tumor Suppressor Genes; Up-Regulation; Viola; Western Blotting; Woman; Xenograft procedure; anticancer research; base; cancer diagnosis; cancer therapy; chemotherapeutic agent; chromatin modification; clinical translation; clinically relevant; curative treatments; disease phenotype; epigenetic regulation; epigenetic silencing; epigenome; genomic locus; human disease; improved; in vivo; inhibitor/antagonist; innovation; mRNA Expression; men; neoplastic cell; novel; recruit; therapeutic gene; tumor; vector

ABSTRACT The disruption of epigenetic pathways are key driving mechanisms that contribute to myriad human diseases. Recent advances in chromatin biology and high-throughput genetic sequencing have discovered that such disruptions underlie a substantial number of cancers. While the affected epigenetic pathways that contribute to cancer pathophysiology are quite diverse, a common theme among them is that aberrant regulation of epigenetic enzymes can lead to dysregulated expression of key disease driver genes. For example, aberrant DNA methylation, histone H3 methylation and/or histone H3 deacetylation can repress genes, and lead to a more deleterious disease phenotype. Indeed, broad-acting pan-epigenetic inhibitors have shown initial promise clinically by grossly disrupting disease signaling pathways by altering the expression profile of key genes. However, a fundamental issue related to the mechanism of these traditional epigenetic inhibitors centers on their potential to affect thousands of genes simultaneously (both on- and off-target). Thus, while treatment with histone deacetylase (HDAC) inhibitors causes the desired effect of activation of specific target genes, this activation also comes with off-target activation of potentially hundreds to thousands of additional genes. Recently, technological advancements on the CRISPR/Cas9 system have been developed to allow for induction of site-specific chromatin modifications that can modulate gene expression. Our technological advancements build on these approaches and allow for the possibility of developing targeted, epigenetically based gene therapeutics with real potential for clinical translation. Here, we outline an approach that leverages a site-specific dCas9-FKBP fusion protein, coupled with a synthetic bifunctional chemical epigenetic modifier (CEM). The CEM consists of three modular components: (1) FK506 (which binds FKBP); (2) a short inert chemical polyethylene glycol (PEG) linker; and (3) a chemical entity that interacts with host epigenetic machinery. Ultimately, the dCas9-FKBP CEM has the ability to target any locus in the genome, and “activate” epigenetic activity to modulate gene expression. We propose to implement this strategy to a clinically relevant target: TP53. Our proposed technology seeks to target endogenous histone acetylation enzymes to the TP53 locus to reverse its epigenetic repression, and thus increase the sensitivity of tumor cells to chemotherapeutic agents. As a first step, we propose to evaluate our technology in preclinical models of colorectal cancer. Colorectal cancer is the third most commonly diagnosed cancer and third cancer most common cause of cancer-related death among both men and women. Additionally, colorectal cancer is known to be driven by both mutated and epigenetically silenced TP53, and there are available preclinical model systems that recapitulate was is observed clinically. Long-term our novel platform could represent an innovative and curative treatment for many epigenetically driven human cancers.

抽象的 表观遗传途径的破坏是导致多种人类疾病的关键驱动机制。 染色质生物学和高通量基因测序的进展发现，这种破坏是 虽然影响癌症病理生理学的表观遗传途径相当多。 多种多样，其中一个共同的主题是表观遗传酶的异常调节可能导致表达失调 关键疾病驱动基因，例如异常 DNA 甲基化、组蛋白 H3 甲基化和/或组蛋白 H3。 脱乙酰化可以抑制基因，并导致更有害的疾病表型，实际上具有广泛的作用。 抑制剂通过改变表达来严重破坏疾病信号传导途径，在临床上显示出初步的希望 然而，一个基本问题与这些传统表观遗传抑制剂中心的机制有关。 因此，在用组蛋白治疗时，它们同时影响数千个基因（目标和脱靶）的潜力。 脱乙酰酶（HDAC）抑制剂会引起特定靶基因激活的预期效果，这种激活也随之而来 最近，技术进步可能导致数百至数千个额外基因的脱靶激活。 CRISPR/Cas9 系统上的修饰已被开发出来，可以诱导位点特异性染色质修饰，从而可以 我们的技术进步建立在这些方法的基础上，并提供了可能性。 开发具有临床转化潜力的靶向、基于表观遗传学的基因疗法。 在这里，我们概述了一种利用位点特异性 dCas9-FKBP 融合蛋白以及合成的 双功能化学表观遗传修饰剂 (CEM) 由三个模块组成：(1) FK506（结合）。 FKBP)；(2) 短惰性化学聚乙二醇 (PEG) 连接体；以及 (3) 与宿主相互作用的化学实体； 最终，dCas9-FKBP CEM 能够靶向基因组中的任何位点并“激活”。 我们建议将这一策略应用于临床相关靶标：TP53。 我们提出的技术旨在将内源性组蛋白乙酰化酶靶向 TP53 基因座，以逆转其表观遗传 抑制，从而增加肿瘤细胞对化疗药物的敏感性作为第一步，我们建议进行评估。 我们的结直肠癌临床前模型技术是第三大最常见的癌症。 第三种癌症是男性和女性癌症相关死亡的最常见原因。 已知是由突变和表观遗传沉默的 TP53 驱动的，并且有可用的临床前模型系统 从长远来看，我们的新平台可以代表一种创新和治疗方法。 治疗许多表观遗传驱动的人类癌症。