Common hTERT Promoter Mutations Represent a Novel Therapeutic Target

常见的 hTERT 启动子突变代表了一个新的治疗靶点

• 批准号: 9225184
• 负责人: Donald M. Miller
• 金额: $ 20.1万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-02-15 至 2018-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9225184
• 关键词: Animal Model; Binding; Biophysics; Catalytic Domain; Cell Proliferation; Cells; Chromosomal translocation; Clinical; Clinical Trials; Down-Regulation; Enzymes; Functional disorder; GTP-Binding Protein alpha Subunits, Gs; Gene Expression; Gene Silencing; Genes; Genetic Transcription; Glioblastoma; Goals; Growth; Human; Invaded; Malignant - descriptor; Mutate; Mutation; Oligonucleotides; Patients; Pharmaceutical Preparations; Physiological; Play; RNA-Directed DNA Polymerase; Regulation; Role; Site; Specificity; Structure; Telomerase; Testing; Therapeutic; Transcriptional Regulation; analytical ultracentrifugation; cancer cell; cancer therapy; cell growth; cell transformation; chemotherapy; design; experience; experimental study; interest; malignant phenotype; melanoma; mutant; new therapeutic target; novel; overexpression; promoter; public health relevance; quadruplex DNA; synergism; targeted agent; targeted treatment; tumor; tumor growth

 DESCRIPTION (provided by applicant): Telomerase reverse transcriptase (hTERT) is a catalytic subunit of the enzyme telomerase which is frequently overexpressed in human tumors. It has recently been shown that the hTERT promoter is commonly mutated in a malignant melanoma and glioblastoma (>75% of tumors). These mutations occur at four specific sites in a G-rich region which has been shown to form quadruplex DNA and to downregulate hTERT expression. We have shown that these mutations destabilize quadruplex-formation resulting in increased hTERT expression and cellular proliferation. Stabilization of the quadruplex structure with TMPYP4 reverses these changes. Treatment of cells in which the hTERT promoter is mutated with "strand-invading" oligonucleotides downregulates hTERT expression and dramatically inhibits the growth of transformed cells. This application will test the hypothesis tht stabilization of the mutated hTERT promoter quadruplex structure by sequence specific oligonucleotides will result in downregulation of hTERT and inhibition of growth by cells with the mutated sequence(s). The Specific Aims of this application are: 1. To characterize the gene and sequence specificity of oligonucleotides targeted to the mutated or wild type hTERT promoter sequences. 2. To characterize the potential synergy of strand invading hTERT promoter targeted oligonucleotides with targeted agents and standard chemotherapy drugs. 3. To characterize the therapeutic potential of hTERT strand invading oligonucleotides in animal models, alone and in conjunction with targeted therapies. The results of the proposed studies will have important clinical implications as inhibition of hTERT gene expression is relevant to a wide variety of tumor types. In addition, the proposed experiments will provide important new information about the role of quadruplex-formation in the regulation of hTERT gene expression.

 描述（由申请人提供）： 端粒酶逆转录酶（hTERT）是端粒酶的催化亚基，其在人类肿瘤中经常过度表达。最近表明，hTERT启动子在恶性黑色素瘤和胶质母细胞瘤（>75）中通常发生突变。这些突变发生在富含 G 的区域的四个特定位点，该区域已被证明可形成四链体 DNA 并下调。我们已经证明，这些突变使四链体形成不稳定，导致 hTERT 表达和细胞增殖增加。用“链侵入”寡核苷酸处理 hTERT 启动子突变的细胞可以逆转这些变化。下调 hTERT 表达并显着抑制转化细胞的生长 该应用将测试突变的 hTERT 启动子四链体结构的稳定性假设。通过序列特异性寡核苷酸进行的修饰将导致具有突变序列的细胞的hTERT下调和生长抑制。本申请的具体目的是： 1.表征针对突变或野生型的寡核苷酸的基因和序列特异性。 hTERT 启动子序列 2. 表征 hTERT 启动子靶向寡核苷酸与靶向药物和标准化疗药物的潜在协同作用 3. 表征 hTERT 链入侵的治疗潜力。动物模型中的寡核苷酸，无论是单独使用还是与靶向治疗相结合，都将具有重要的临床意义，因为 hTERT 基因表达的抑制与多种肿瘤类型相关。此外，所提出的实验将提供重要的新内容。有关四链体形成在 hTERT 基因表达调节中的作用的信息。

项目成果

Quadruplex-forming oligonucleotide targeted to the VEGF promoter inhibits growth of non-small cell lung cancer cells.

靶向 VEGF 启动子的四链体形成寡核苷酸可抑制非小细胞肺癌细胞的生长。

• 发表时间: 2019
• 影响因子: 3.7
• 作者: Muench, David;Rezzoug, Francine;Thomas, Shelia D;Xiao, Jingjing;Islam, Ashraful;Miller, Donald M;Sedoris, Kara C
• 通讯作者: Sedoris, Kara C