Biomodulation of anticancer drugs targeting DNA

靶向DNA的抗癌药物的生物调节

• 批准号: 7474048
• 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/7474048
• 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，TSA或SANA，两种组蛋白去乙酰化酶抑制剂(HDACIs)预先处理几种癌细胞株，可以增加VP-16，椭圆，阿霉素和顺铂的杀伤率。由于在正常细胞中没有观察到致敏作用，或者当在癌细胞中抗癌药物之后而不是在抗癌药物之前加入HDACIs时，我们假设正常细胞和癌细胞染色质之间的内在差异允许HDACIs增加抗癌药物在癌细胞中的DNA可及性。具体目的：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，来自同步细胞的染色质将被消化，并将测量核小体重复长度。组蛋白H1的磷酸化水平以及异染色质蛋白HP1a和组蛋白H4在Lys16乙酰化的分布也将被评估为染色质紧凑的指标。2)确定HDACIs对药物可及性的影响是否与正常细胞和癌细胞的染色质不同。这将通过一种改进的芯片试验来执行，以测量抗癌药物靶标DNA序列附近的染色质可及性，并通过PCR-STOP试验进行。3)确定在癌细胞中过度表达的组蛋白结合蛋白是否参与了HDACIs的增敏作用。我们将评估HMG-I/Y对针对DNA或作用于DNA的酶的抗癌药物增敏HDACIs的潜在作用。这将通过下调癌细胞中hMG-I/Y的水平和确定足以介导这一效应的结构域(S)来实现。意义：我们的初步研究表明，用HDACIs对癌细胞进行预处理可以提高抗癌药物的杀伤率。2005年11月，我们机构批准了一期临床试验，将这项研究扩大到复发性和/或急性白血病和骨髓增生异常综合征患者。这一建议将有助于更好地了解介导这一效应的基本机制，并将有助于指导和发展癌症治疗的基于机制的疗法。