Chromatin Organization and Nuclear compartmentalization in Osteoblast Gene Exp...

成骨细胞基因实验中的染色质组织和核区室化...

基本信息

批准号：
8289360
负责人：
Janet L Stein
金额：
$ 38.85万
依托单位：
UNIV OF MASSACHUSETTS MED SCH WORCESTER
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-01 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8289360
关键词：
Address Adult Affect Antibodies Autoradiography Award Binding Binding Proteins Biological Bone Diseases Budgets Calvaria Candidate Disease Gene Categories Cell Line Cell Nucleus Cell model Cells Cellular biology Chemicals Chile Cholecalciferol Chromatin Chromatin Remodeling Factor Chromatin Structure Clinical Collaborations Complement Complex Cues Development Differentiation and Growth Direct Costs Disease EP300 gene Electrophoresis Enzymes Epigenetic Process Equipment Event Face Facilities and Administrative Costs Fees Funding Gene Activation Gene Expression Gene Expression Profile Gene Expression Regulation Gene Structure Gene Targeting Genes Genetic Transcription Goals Histone Code Histone Deacetylase Inhibitor Histones Human Resources Immunology Inpatients Instruction Interphase Isotopes Laboratories Last Name Maintenance Massachusetts Mature Bone Mediating Medical Metabolic Mitosis Modeling Modification Molecular Molecular Biology Names Nuclear Nuclear Matrix Nuclear Matrix-Associated Proteins Null Lymphocytes Osteoblasts Osteocalcin Osteogenesis Outpatients Patient Care Pattern Persons Phenotype Physiological Plastics Play Point Mutation Principal Investigator Program Research Project Grants Publications Radioisotopes Rattus Reagent Recruitment Activity Registries Research Research Personnel Role Signal Pathway Signal Transduction Site Skeletal Development Skeleton Staging Students Therapeutic Intervention Time Tissues Transcriptional Activation Transcriptional Regulation Universities VDR gene Vitamin D Vitamin D3 Receptor Vitamins bioimaging bone bone cell bone quality bone strength bone turnover chromatin remodeling clinically relevant cost genetic regulatory protein histone modification human embryonic stem cell in vivo indexing inhibitor/antagonist insight knock-down medical schools meetings novel nuclease osteoblast differentiation osteogenic programs promoter receptor response steroid hormone transcription factor

项目摘要

A fundamental problem in bone cell biology, is how bone-specific genes are rendered competent for expression and steroid hormone responsiveness at appropriate stages during the osteoblast developmental sequence. This program established that Runx2, the key transcription factor required for bone formation, plays a critical role in the assembly of active chromatin at the osteocalcin (OC)promoter during phenotype progression and is required for steroid hormone responsiveness. The long term goal of this project is to understand how dynamic reorganization of the chromatin of bone-specific genes at subnuclear foci contributes to activation of the osteoblast developmental transcription program in response to physiologic regulatory cues. Importantly, during the current award period, we have demonstrated that activity of the SWI/SNF chromatin remodeling complex is also essential both for expression and vitamin D enhancement of the OC gene in osteoblastic cells and for osteoblast differentiation. We now propose to extend our exciting findings to address the hypothesis that induction of the osteoblast phenotvpe must involve histone modifications and ordered remodeling of chromatin in an osteoblast-specific manner in response to tissue- specific factors and multi-component enzymatic complexes that bind sequentially to activate and regulate transcription and mediate vitamin D responsiveness. To understand more broadly how gene expression is regulated during skeletal development, we will pursue a highly collaborative approach to characterize: 1. the Runx2-dependent and vitamin D responsive program of binding events and chromatin remodeling at the osteocalcin promoter; 2. epigenetic marking and chromatin remodeling of bone phenotypic genes in response to Runx2 and vitamin D during osteoblast differentiation; and 3. the Runx2 and Vitamin D responsive gene expression program required for establishment and maintenance of the mature bone cell phenotype. These studies will reveal the role of Runx2 and key co-factors for developmental expression and vitamin D dependent regulation of genes required for osteoblast differentiation. Collaborations with Projects 1 and 2 will provide novel insights into the function of Runx2 in supporting local architectural organization of gene promoters and co-regulators for bone-specific transcriptional control. Clinical Relevance: Defining mechanisms that control chromatin organization is compelling for understanding actions and consequences of inhibitors that target chromatin for the treatment of diseases that affect bone turnover in the adult skeleton. University of Massachusetts Medical School 55 Lake Avenue North Worcester, MA 01655 PHS 398 (Rev. 04/06) Page 240 Form Page 2 Project 3 Principal Investigator/Program Director (Last, First, Middle): Stein, Gary S. KEY PERSONNEL. See instructions. Use continuation pages as needed to provide the required information in the format shown below. Start with Principal Investigator(s). List all other key personnel in alphabetical order, last name first. Name eRA Commons User Name Organization Role on Project Stein, Janet L. JLSTEIN UMASS Medical Sch. Principal Investigator Montecino, Martin mmonteci Universidad de Co-Pi Concepcion, Chile Imbalzano, Anthony N. AN IMBALZANO UMASS Medical Sch. Co-Pi OTHER SIGNIFICANT CONTRIBUTORS Name Organization Role on Project Lian, Jane B. UMASS Medical Sch. Collaborator Stein, Gary S. UMASS MedicalSch. Collaborator van Wijnen, Andre J. UMASS Medical Sch. Collaborator Lapointe, David UMASS Medical Sch. Collaborator Human Embryonic Stem Cells [X] No D Yes If the proposed project involves human embryonic stem cells, list below the registration number of the specific cell llne(s) from the following list: http://stemcells.nih.qov/registrv/index.asp. Use continuation pages as needed. If a specific line cannot be referenced at this time, include a statement that one from the Registry will be used. Cell Line PHS 398 (Rev. 04/06) Page 241 Form Page 2-continued Number the followingpages consecutively throughout the application. Do not use suffixes such as 4a, 4b. PROJECT 3 Principal Investigator/Program Director (Last, First, Middle): Stein, Gary S. DETAILED BUDGET FOR INITIAL BUDGET PERIOD FROM THROUGH DIRECT COSTS ONLY 7/1/07 6/30/08 PERSONNEL (Applicant organization only) Months Devoted to Project ROLE ON Cal. Acad. Sum. INST.BASE NAME PROJECT Mnths Mnths Mnths SALARY Principal Janet L. Stein 1.80 166,683 Investigator Co-Principal Martin Montecino 1.20 0 Investigator Co-Principal Anthony Imbalzano 1.20 126,764 Investigator Research Thomas Barthel Associate 12.00 38,000 Postdoctoral Sharanjot Saini 12.00 36,000 Associate Graduate Cara Weismann 12.00 25,740 Student SUBTOTALS CONSULTANT COSTS EQUIPMENT (Itemize) DOLLAR AMOUNT REQUESTED (omit cents) SALARY FRINGE REQUESTED BENEFITS TOTAL 25,002 8,686 33,688 0 0 0 12,676 4,404 17,080 38,000 13,201 51,201 36,000 12,506 48,506 25,740 3,573 29,313 137,418 42,370 179,788 SUPPLIES (Itemize by category) Radioisotopes and autoradiography 5,400 General chemicals 7,000 Molecular biology enzymes and kits 13,000 Antibodies/Immunology 9,300 Electrophoresis reagents/supplies 5,100 Laboratory supplies and plastics 10,437 50,237 TRAVEL ASBMR Annual Meeting (3 personnel), 4,800; M. Montecino (4 trips/year), 8,000 12,800 PATIENT CARE COSTS INPATIENT OUTPATIENT ALTERATIONS AND RENOVATIONS (Itemize by category) OTHER EXPENSES (Itemize by category) Isotope fee/person (3 x 350) 1 ,050 Equipment maintenance 2,500 Publication costs 3,350 Biomedical imaging 2,000 8,900 CONSORTIUM/CONTRACTUAL COSTS DIRECT COSTS SUBTOTAL DIRECT COSTS FOR INITIAL BUDGET PERIOD (Item 7a, Face Page) $ 251 ,725 CONSORTIUM/CONTRACTUAL COSTS FACILITIES AND ADMINISTRATIVE COSTS TOTAL DIRECT COSTS FOR INITIAL BUDGET PERIOD $ 251,725 PHS 398 (Rev.04/06) Page 242 Form Page 4

骨细胞生物学中的一个基本问题是如何使骨特异性基因具有胜任在成骨细胞发育期间，在适当阶段的表达和类固醇激素反应性顺序。该程序确定runx2是骨形成所需的关键转录因子，在表型期间在骨钙蛋白（OC）启动子的活性染色质组装中起关键作用进展，是类固醇激素反应性所必需的。该项目的长期目标是了解如何在亚核灶处的骨特异性基因染色质的动态重组有助于激活成骨细胞发育转录程序，以响应生理监管提示。重要的是，在当前奖励期间，我们已经证明了 SWI/SNF染色质重塑复合物对于表达和维生素D的增强也至关重要成骨细胞中的OC基因和成骨细胞分化。我们现在建议扩展我们的激动人心解决成骨细胞示威的假设的发现必须涉及组蛋白对组织的修饰和以成骨细胞特异性方式重塑染色质特定因素和多组分酶络合物，依次结合以激活和调节转录并介导维生素D的反应性。更广泛地了解基因表达方式在骨骼发育期间受到监管，我们将采取一种高度协作的表征：1。 runx2依赖性和维生素D的结合事件和染色质重塑的反应敏感程序骨钙蛋白启动子； 2。骨表型基因的表观遗传标记和染色质重塑在成骨细胞分化过程中对Runx2和维生素D的反应；和3。runx2和维生素D 建立和维持成熟的骨细胞所需的反应敏感基因表达程序表型。这些研究将揭示runx2和关键的共同因素在发育表达和维生素D的依赖性调节成骨细胞分化所需的基因。与之合作项目1和2将提供有关Runx2在支持本地建筑的功能的新见解基因启动子和骨特异性转录控制的共同调节器的组织。临床相关性：控制染色质组织的定义机制令人信服靶向染色质的抑制剂的作用和后果，以治疗影响骨骼的疾病成人骨骼的营业额。马萨诸塞大学医学院北湖大道55号伍斯特，马萨诸塞州01655 PHS 398（修订版04/06）第240页表格2 项目3 首席调查员/计划主管（最后，第一，中间）：斯坦因，加里·S。关键人员。请参阅说明。根据需要使用延续页面，以下面显示的格式提供所需的信息。从首席研究员开始。按字母顺序列出所有其他关键人员，首先姓氏。名称ERA CONSONS用户名组织在项目中的角色 Stein，Janet L. Jlstein Umass Medical Sch。首席研究员 Montecino，Martin Mmonteci De Co-Pi大学 Concepcion，智利 Imbalzano，Anthony N. Imbalzano Umass Medical Sch。 Co-Pi 其他重要的贡献者姓名组织角色 Lian，Jane B. Umass Medical Sch。合作者 Stein，Gary S. Umass Medicalsch。合作者 Van Wijnen，Andre J. Umass Medical Sch。合作者 Lapointe，David Umass Medical Sch。合作者人类胚胎干细胞[x]否是如果提出的项目涉及人类胚胎干细胞，则从以下列表中列出了特定细胞LLNE的注册号以下： http：//stemcells.nih.qov/registrv/index.asp。根据需要使用延续页面。如果目前无法引用特定行，请包括一条说明将使用注册表的声明。细胞系 PHS 398（修订版04/06）第241页表格第2页。在整个过程中连续编号应用程序。请勿使用4A，4B等后缀。项目3首席调查员/计划主管（最后，第一，中间）：Stein，Gary S. 最初预算期的详细预算从直接费用仅7/1/07 6/30/08 人员（仅申请人组织）致力于项目的月份在Cal上的角色。学院。和。 Inst.Base 名称项目MNTHS MNTHS MNTHS工资主要的 Janet L. Stein 1.80 166,683 研究者联合主持 Martin Montecino 1.20 0 研究者联合主持 Anthony Imbalzano 1.20 126,764 研究者研究托马斯·巴瑟（Thomas Barthel）助理12.00 38,000 博士后 Sharanjot Saini 12.00 36,000 联系毕业 Cara Weismann 12.00 25,740 学生小计顾问费用设备（逐项）要求的美元金额（省略美分）薪水附带请求的福利总计 25,002 8,686 33,688 0 0 0 12,676 4,404 17,080 38,000 13,201 51,201 36,000 12,506 48,506 25,740 3,573 29,313 137,418 42,370 179,788 供应（按类别逐项列出）放射性同位素和放射自显影5,400个通用化学物质7,000 分子生物学酶和试剂盒13,000抗体/免疫学9,300 电泳试剂/供应5,100 实验室用品和塑料10,437 50,237 旅行 ASBMR年会（3人），4,800； M. Montecino（4次旅行/年），8,000 12,800 患者护理费用住院门诊更改和翻新（按类别逐项列出）其他费用（按类别逐项列出）同位素费/人（3 x 350）1，050 设备维护2,500 出版物成本为3,350 生物医学成像2,000 8,900 财团/合同费用直接费用初始预算期（项目7a，face Page）的小计直接费用$ 251，725 财团/合同费用设施和行政费用初始预算期间的总直接成本$ 251,725 PHS 398（Rev.04/06）Page 242表格Page 4