Development of a cavitation enhancement technology to access archived tissues for epigenetic-based biomedical research

开发空化增强技术以获取存档组织以进行基于表观遗传学的生物医学研究

基本信息

批准号：
10211263
负责人：
Ian J Davis
金额：
$ 40.15万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-04-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10211263
关键词：
Acoustics Address Architecture Biological Biological Assay Biology Biomedical Research Cell Nucleus Cells Cellular Assay Chemicals Chromatin Consumption DNA DNA Fragmentation DNA Packaging DNA-Protein Interaction Data Dehydration Development Devices Diagnostic Disease Disease Resistance Enhancement Technology Epigenetic Process Epilepsy Formaldehyde Formalin Formulation Genomic DNA Goals Heating Heterogeneity Histones Human Human Development Intellectual functioning disability Legal patent Link Malignant Neoplasms Methods Mutation Nuclear Proteins Nucleic Acids Nucleosomes Paraffin Embedding Pattern Proteins Protocols documentation Reagent Regulation Regulatory Pathway Reproducibility Research Sampling Sonication Specimen Standardization Syndrome Techniques Technology Time Tissue Banks Tissue Embedding Tissue Sample autism spectrum disorder base biological specimen archives cell type chromatin immunoprecipitation crosslink driver mutation human disease innovation nanoDroplet nervous system disorder new technology next generation sequencing parallel processing preservation prevent programs sample fixation therapy resistant tissue archive tumor virtual

项目摘要

Project Summary There is a growing appreciation for the importance of epigenetic regulatory mechanisms in both normal human development and disease. The dynamic regulation of chromatin architecture is a frequently studied and central epigenetic process. Mutations in chromatin-associated proteins have been linked to neurological disorders such as intellectual disability, autism, and epilepsy, a variety of developmental syndromes, and detected in approximately 50% of human cancers, many of which are recognized as driver mutations across multiple diverse tumors. Nevertheless, many of the assays used to study chromatin biology remain cumbersome, expensive, or lack experimental robustness, problems which have resulted in large gaps in our understanding of this critical biological mechanism. Therefore, innovative new technologies that enable efficient and reproducible interrogation of epigenetic mechanisms are badly needed. We have invented a unique cavitation enhancement reagent that dramatically decreases the time and acoustic energy required for genomic DNA fragmentation in a sonication device. This proposal aims to expand the application of our cavitation enhancement technology to address the scientifically relevant challenge of extracting high quality chromatin from archival tissues. Lengthy formaldehyde fixation (FF) followed by dehydration and paraffin embedding (PE) is the standard method of archiving tissue samples. These FFPE tissue collections contain a wealth of information on human disease. However, extraction of chromatin from these specimens has proven virtually impossible because of the challenge of preserving relevant DNA-protein interactions. Preliminary data suggest that our cavitation enhancement reagent has the potential to simplify and standardize chromatin extraction from archived tissues, thereby greatly expanding the range of applications for FFPE-derived chromatin in epigenetic-based biomedical research.

项目摘要人们对表观遗传调节机制的重要性越来越欣赏正常人发展与疾病。染色质结构的动态调节是经常研究的中心表观遗传过程。染色质相关蛋白的突变与神经系统疾病有关作为智力残疾，自闭症和癫痫，多种发育综合症，并在大约50％的人类癌症，其中许多被认为是多种多样的驾驶员突变肿瘤。然而，许多用于研究染色质生物学的测定方法仍然繁琐，昂贵或缺乏实验性鲁棒性，问题导致我们对这一关键的理解差距很大生物机制。因此，创新的新技术可以有效且可重复急需对表观遗传机制的询问。我们发明了一种独特的空化增强试剂，该试剂大大减少了时间和声学基因组DNA片段化所需的能量。该建议旨在扩大我们的空化增强技术的应用来应对提取的科学相关挑战档案组织的高质量染色质。冗长的甲醛固定（FF），然后脱水和石蜡嵌入（PE）是存档组织样品的标准方法。这些FFPE组织收集包含大量有关人类疾病的信息。但是，从这些标本中提取染色质由于维护相关的DNA蛋白质相互作用的挑战，实际上几乎是不可能的。初步的数据表明，我们的空化增强试剂具有简化和标准化染色质的潜力从存档的组织中提取，从而大大扩展了FFPE衍生染色质的应用范围在基于表观遗传学的生物医学研究中。