Molecular Sensors for Imaging Histone Methylations in Living Animals

用于活体动物组蛋白甲基化成像的分子传感器

• 批准号: 8635985
• 负责人: Ramasamy Paulmurugan
• 金额: $ 31.66万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8635985
• 关键词: Alanine; Animals; Azacitidine; Behavior; Binding; Biological Assay; Bioluminescence; Cell Culture Techniques; Cell Line; Cells; DNA Methylation; DNA Sequence; Development; Disease; Drug Evaluation; Drug Kinetics; Drug Targeting; Drug usage; Epigenetic Process; Event; Firefly Luciferases; Functional disorder; Gene Expression; Goals; Grant; Histone Acetylation; Histones; Image; Lead; Leucine; Life; Link; Luciferases; Lysine; Malignant Neoplasms; Mammalian Cell; Measures; Methylation; Methyltransferase; Modification; Molecular; Monitor; Mus; N-terminal; Neoplasm Metastasis; Optics; Peptides; Pharmaceutical Preparations; Pharmacodynamics; Phenotype; Positron-Emission Tomography; Preclinical Drug Evaluation; Process; Protein Fragment; Proteins; Renilla; Reporter; Research; Research Proposals; Role; Specificity; System; Techniques; Testing; Therapeutic; Therapeutic Studies; Thymidine Kinase; Tretinoin; Trichostatin A; Tryptophan; Valproic Acid; Vorinostat; Work; analytical tool; base; bioluminescence imaging; cellular imaging; clinical application; combinatorial; drug development; flexibility; histone methyltransferase; imaging modality; improved; in vivo; in vivo imaging; inhibitor/antagonist; innovation; interest; molecular imaging; mutant; novel; novel strategies; optical imaging; pre-clinical; protein H(3); research clinical testing; response; sensor; small molecule; subcutaneous; tool; tumor xenograft

DESCRIPTION (provided by applicant): "Epigenetics" refers to changes in phenotype or gene expression caused by mechanisms other than changes in the underlying DNA sequence. Abnormal epigenetic control mechanisms are now regarded as significant contributing factors to the pathophysiology of different diseases, including cancer. The main epigenetic mechanisms that induce phenotypic changes in mammalian cells are DNA methylation, histone methylation and histone acetylation. Many recent targeted drug development efforts have been directed towards the modification of one or more of these epigenetic mechanisms. In that regard, several cell culture based assays are being used to analyze different epigenetic processes, and small molecule drugs modulate these epigenetic processes. However these tests are not capable of predicting the exact behavior of drug pharmacokinetics and pharmacodynamics in living subjects. The current intense efforts to develop such drugs, as well as an ever increasing interest in epigenetics research, creates a paramount need to develop new more meaningful molecular imaging strategies that can specifically interrogate different epigenetic mechanisms in vivo. The main goal of this proposal is to develop novel in vivo imaging strategies to monitor cellular epigenetic processes in living animals, evaluating these techniques when imaging the basic epigenetic shifts occurring in the development of different cancers, and to apply these imaging platforms to study therapeutic drugs that modulate these processes in cancer. Specifically, we wish to image and quantitate histone methylation in vivo, and to apply this novel analytical tool to evaluate drugs that modulate histone methylation, which may have important applications in molecular therapeutics of cancer. This will be achieved by developing: 1) Optical bioluminescence (Split-Luciferase-complementation), and 2) microPET (Split-Thymidine kinase complementation) imaging sensors. Combinatorial therapies using different epigenetic modulators (inhibitors of histone deacetylases in combination with histone methyltransferases) is curently considered as a new approach for treating cancers and several other intractable cellular diseases. The sensors we are planning to develop by this grant, are having the potential to image molecular events in both cells and in live animals. These sensors will improve the use of epigenetic modulators in translational clinical applications by enabling drug screening and their pre-clinical evaluations in living animals. The aberrant histone methylation has been considered as an important player in the development of cancer. In summary, this proposal will lead to the development of highly sensitive in vivo imaging methods that can be used to further epigenetic research, as well as accelerating the pre-clinical evaluation of drugs targeting different cancers and other cellular diseases.

描述（由申请人提供）：“表观遗传学”是指表型或基因表达的变化，其由基础DNA序列变化以外的机制引起。异常的表观遗传控制机制现在被认为是不同疾病（包括癌症）的病理生理学的重要贡献因素。诱导哺乳动物细胞表型改变的主要表观遗传机制是DNA甲基化、组蛋白甲基化和组蛋白乙酰化。许多最近的靶向药物开发工作已经针对这些表观遗传机制中的一个或多个的修饰。在这方面，几种基于细胞培养的测定法被用于分析不同的表观遗传过程，并且小分子药物调节这些表观遗传过程。然而，这些试验不能预测活体受试者中药物药代动力学和药效学的确切行为。目前开发此类药物的紧张努力，以及对表观遗传学研究的日益增长的兴趣，产生了开发新的更有意义的分子成像策略的迫切需要，这些策略可以特异性地询问体内不同的表观遗传机制。该提案的主要目标是开发新的体内成像策略，以监测活体动物中的细胞表观遗传过程，在对不同癌症发展中发生的基本表观遗传变化进行成像时评估这些技术，并将这些成像平台应用于研究调节癌症这些过程的治疗药物。具体来说，我们希望在体内成像和定量组蛋白甲基化，并应用这种新的分析工具来评估药物，调节组蛋白甲基化，这可能有重要的应用在癌症的分子治疗。这将通过开发：1）光学生物发光（分裂荧光素酶互补）和2）microPET（分裂胸苷激酶互补）成像传感器来实现。使用不同表观遗传调节剂（组蛋白去乙酰化酶抑制剂与组蛋白甲基转移酶组合）的组合疗法目前被认为是治疗癌症和其他几种难治性细胞疾病的新方法。我们计划利用这笔资金开发的传感器，有可能在细胞和活体动物中成像分子事件。这些传感器将通过在活体动物中进行药物筛选和临床前评估来改善表观遗传调节剂在翻译临床应用中的使用。组蛋白甲基化异常被认为是肿瘤发生发展的重要因素。总之，该提案将导致开发高灵敏度的体内成像方法，可用于进一步的表观遗传学研究，以及加速针对不同癌症和其他细胞疾病的药物的临床前评估。