Biomodulation of anticancer drugs targeting DNA

靶向DNA的抗癌药物的生物调节

• 批准号: 7259571
• 负责人: France Carrier
• 金额: $ 22.8万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7259571
• 关键词: Active Sites; Acute leukemia; Address; Affect; Affinity; Aftercare; Antibodies; Antineoplastic Agents; Apoptosis; Architecture; Binding; Binding Proteins; Binding Sites; Biological Assay; Cancer cell line; Cell Cycle; Cell Extracts; Cell Line; Cell physiology; Cells; Chromatin; Cisplatin; Classification; Clinical Trials Design; Code; Condition; DNA; DNA Binding Domain; DNA Damage; DNA Packaging; DNA Sequence; Data; Decompression Sickness; Diagnosis; Digestion; Drug Delivery Systems; Ellipticines; Enzymes; Exons; France; Gel; Genes; Goals; HMGA1 gene; Heterochromatin; Histone Deacetylase Inhibitor; Histone H1; Histone H1(s); Histone H3; Histone H4; Immunoprecipitation; Institution; Length; Localized; MCF7 cell; Malignant Neoplasms; Measurement; Measures; Mediating; Menotropins; Micrococcal Nuclease; Modification; Molecular; Monitor; Morphology; Normal Cell; Nuclear; Nucleosomes; Oncogenes; Patients; Pattern; Pharmaceutical Preparations; Phase; Phase I Clinical Trials; Phosphorylation; Pliability; Point Mutation; Polymerase Chain Reaction; Post-Translational Protein Processing; Protein Overexpression; Proteins; Rate; Relapse; Research Personnel; Site; Small Interfering RNA; Southern Blotting; Staphylococcal Protein A; Structure; Superhelical DNA; Syndrome; Testing; Therapeutic; Topoisomerase; Transfection; Trichostatin A; U118; Vorinostat; base; c-myc Genes; cancer cell; cancer therapy; carcinogenesis; chromatin immunoprecipitation; cisplatin/doxorubicin protocol; cytotoxic; day; drug discovery; ellipticine; heterochromatin-specific nonhistone chromosomal protein HP-1; histone-binding proteins; human H2AX protein; hydroxymethylglutarate; improved; killings; mutant; neoplastic cell; prevent; programs; promoter; succinyl-trialanine-4-nitroanilide; transcription factor

DESCRIPTION (provided by applicant): Hypothesis: We have recently shown that pre-treatment of several cancer cell lines with TSA or SANA, two Histone Deacetylase Inhibitors (HDACIs), increased the killing efficiency of VP-16, ellipticine, doxorubicin and cisplatin. Because no sensitizing effect was observed in normal cells or when the HDACIs were added after instead of before the anticancer drugs in cancer cells, we hypothesized that intrinsic differences between the chromatin of normal and cancer cells allow the HDACIs to increase the DNA accessibility of the anticancer drugs in the cancer cells. Specific Aims: To verify this hypothesis we will: 1) Determine if the chromatin compaction of normal and cancer cells is affected differently by the HDACIs. Bulk chromatin will be digested with Micrococcal Nuclease (MNase) and chromatin will also be digested at specific loci targeted by the anticancer drugs. In addition, chromatin from synchronized cells will be digested and the nucleosome repeat length will be measured. Levels of histone H1 phosphorylation and distribution of the heterochromatin protein HP1a and the histone H4 acetylated at Lys16 will also be evaluated as indicators of chromatin compaction. 2) Determine if the effect of HDACIs on drug accessibility is different on the chromatin of normal and cancer cells. This will be performed by a modified Chips assay to measure chromatin accessibility in the vicinity of the DNA sequences targeted by the anticancer drugs and by the PCR-stop assay. 3) Determine if histone binding proteins overexpressed in cancer cells contribute to the sensitizing effect of HDACIs. We will evaluate the potential enhancing effect of HMG-I/Y on HDACIs sensitization to anticancer drugs targeting the DNA or enzymes acting on the DNA. This will be performed by down regulating the levels of HMG-I/Y in cancer cells and by identifying the domain(s) sufficient to mediate this effect. Significance: Our initial study demonstrated that pre-treatment of cancer cells with HDACIs increased the killing efficiency of anticancer drugs. On November 2005, a Phase 1 clinical trial was approved at our institution to expand this study to patients with relapsed and/or acute leukemia and myeolodysplastic syndromes. This proposal will provide a better understanding of the basic mechanisms mediating this effect and will contribute to guide and develop mechanism-based therapeutics for cancer treatments.

描述（由申请人提供）： 假设：我们最近表明，用 TSA 或 SANA（两种组蛋白脱乙酰酶抑制剂 (HDACIs)）预处理几种癌细胞系，可以提高 VP-16、玫瑰树碱、阿霉素和顺铂的杀伤效率。由于在正常细胞中没有观察到敏化作用，或者当 HDACIs 在癌细胞中添加抗癌药物之后而不是之前添加时，我们假设正常细胞和癌细胞染色质之间的内在差异使得 HDACIs 能够增加癌细胞中抗癌药物的 DNA 可及性。具体目标：为了验证这一假设，我们将：1) 确定正常细胞和癌细胞的染色质压缩是否受到 HDACIs 的不同影响。大量染色质将被微球菌核酸酶 (MNase) 消化，染色质也将在抗癌药物靶向的特定位点被消化。此外，同步化细胞的染色质将被消化，并测量核小体重复长度。组蛋白 H1 磷酸化水平以及异染色质蛋白 HP1a 和 Lys16 乙酰化组蛋白 H4 的分布也将作为染色质压缩的指标进行评估。 2) 确定 HDACIs 对药物可及性的影响对正常细胞和癌细胞的染色质是否不同。这将通过改进的 Chips 测定来测量抗癌药物靶向的 DNA 序列附近的染色质可及性，并通过 PCR 终止测定来进行。 3) 确定癌细胞中过度表达的组蛋白结合蛋白是否有助于 HDACIs 的敏化作用。我们将评估 HMG-I/Y 对 HDACIs 对靶向 DNA 或作用于 DNA 的酶的抗癌药物的敏感性的潜在增强作用。这将通过下调癌细胞中的 HMG-I/Y 水平并通过鉴定足以介导这种作用的结构域来实现。意义：我们的初步研究表明，用 HDACIs 预处理癌细胞可提高抗癌药物的杀伤效率。 2005 年 11 月，我们机构批准了一项 1 期临床试验，将这项研究扩展到患有复发和/或急性白血病和骨髓增生异常综合征的患者。该提案将有助于更好地理解介导这种效应的基本机制，并将有助于指导和开发基于机制的癌症治疗方法。