Gravitational Regulation of Osteoblast Genomics and Metabolism

成骨细胞基因组和代谢的重力调节

基本信息

批准号：
8326052
负责人：
BRUCE E HAMMER
金额：
$ 22.68万
依托单位：
UNIVERSITY OF MINNESOTA
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：
2011-09-01 至 2013-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8326052
关键词：
Address Affect American Animals Apoptosis Astronauts Back Beds Biochemical Biological Biological Assay Biological Markers Biological Preservation Bioreactors Bone remodeling Cell Culture System Cell Culture Techniques Cell Differentiation process Cells Cellular Morphology Certification Cues Development Differentiation Antigens Disease Drug Design Environment Enzyme-Linked Immunosorbent Assay Evaluation Force of Gravity Fracture Gene Expression Genes Genomics Habitats Homeostasis Incubators Individual International Logistics MAP Kinase Gene MAPK Signaling Pathway Pathway Magnetism Maintenance Measurable Measures Mechanical Stress Metabolism Methods Metric Microarray Analysis Microgravity Minnesota Mission Molecular NFKB Signaling Pathway NMR Spectroscopy Nutrient Ocular orbit Orbital Simulations Osteoblasts Osteoclasts Osteogenesis Osteonectin Osteopenia Osteoporosis Outcome Pathway interactions Patients Performance Phase Physical therapy Physiology Proteomics Protocols documentation Regulation Research Research Personnel Reverse Transcriptase Polymerase Chain Reaction Role Running Sampling Screening procedure Signal Pathway Signaling Pathway Gene Simulate Stress TNFSF11 gene Techniques Technology Testing Tissue-Specific Gene Expression Training United States National Aeronautics and Space Administration United States National Institutes of Health Universities Water Weight-Bearing state Western Blotting Work base bone bone cell bone loss bone mass cell growth experience falls instrumentation metabolomics older patient osteopontin protein expression research study response skeletal tissue tool

项目摘要

DESCRIPTION (provided by applicant): The role of gravity in the development and maintenance of bone on earth is controversial. Our current understanding is that gravitational unloading of bone results in bone loss (osteopenia) which typically progresses to osteoporosis and ultimately bone fracture. This is a significant problem that bed- ridden patients and the elderly experience. Astronauts in orbit experience significant osteopenia of approximately 1% to 3% of bone loss per month, which is equivalent to a year of bone loss for osteoporotic individuals on earth. Understanding bone response to varying levels of gravitational force is critical toward developing rational approaches in drug design or physical therapy toward mitigating osteopenia. Presently, the only means of exposing bone to a long-term lack of gravitational force is through orbital free fall aboard the ISS. These studies are expensive and logistically difficult to do aboard the ISS. An ideal method of study gravitational regulation of bone dynamics is to have a ground-based method to remove or reduce the gravitational force on bone. Creating an environment that amplifies the rate of bone loss will give us a tool to accelerate our understanding of the underlying genomic, proteomic and metabolomic mechanisms. The proposed study seeks to determine if ground-based magnetic levitation is a suitable simulation of orbital free fall for cell culture studies. Studying a cell culture rather than an entire animal allows for highly controlled and reproducible experiments. We plan to grow MC3T3 osteoblastic and primary osteoclastic cells in a bioreactor compatible with our ground-based levitation magnet and the ISS space-based microgravity environment. Our unique levitation magnet integrates a 37 C incubator integrated into the magnet bore allowing levitation for several weeks. The implementation partner for experiments aboard the ISS is BioServe Space Technologies in Boulder, CO. BioServe has a two-decade record of accomplishment of working with NASA on delivering biological experiments to and from low-earth orbit. Osteoblasts and osteoclasts flown on previous orbital missions show evidence that microgravity affects cell differentiation and gene expression. The metrics for the proposed study include cell morphology studies, microarray analysis to measure differential gene expression, ELISA and RT-PCR to identify markers of osteoblast and osteoclast differentiation, metabolomics, via H-1 NMR spectroscopy and quantification of MAPK, Twsg1 and NF-kB signaling pathways. With this suite of assays, we will determine similarities and differences in a number of metabolites and biomarkers for simulated microgravity and ISS microgravity environments. This project will help identify mechanism(s) and pathways that can be addressed to mitigate bone loss on earth.

描述（由申请人提供）：重力在地球上骨骼的发展和维持中的作用是有争议的。我们目前的理解是，骨骼的重力卸载会导致骨质流失（骨质减少），这通常会发展为骨质疏松症并最终导致骨折。这是一个重大的问题，即骑患者和老年人经历。轨道上的宇航员每月的骨质减少症约为骨质损失的1％至3％，这相当于地球上骨质疏松个体的骨质流失一年。了解骨对不同水平的重力反应对于发展药物设计或物理治疗中的理性方法至关重要。目前，将骨骼暴露于长期缺乏引力力的唯一手段是通过ISS上的无轨道秋天。这些研究很昂贵，在ISS上很难进行。研究骨动力学的重力调节的理想方法是采用基于地面的方法来去除或减少骨头上的重力。创造一个扩大骨质流失率的环境将为我们提供一种工具，以加速我们对潜在的基因组，蛋白质组学和代谢组机制的理解。拟议的研究旨在确定地面磁悬浮是否是对细胞培养研究的轨道无轨道下落的合适模拟。研究细胞培养而不是整个动物可以进行高度控制和可重复的实验。我们计划在生物反应器中与我们的地面悬浮磁体和ISS空间基的微重力环境兼容MC3T3成骨细胞和原代成骨细胞。我们独特的悬浮磁铁将整合到磁铁孔中的37 C孵化器整合，允许悬浮数周。 ISS上的实验伙伴是ISS的BioServe太空技术。BioServe拥有与NASA合作进行生物学实验往返低年度轨道的两年成绩的两年记录。在先前的轨道任务上飞行的成骨细胞和破骨细胞显示了微重力影响细胞分化和基因表达的证据。拟议研究的指标包括细胞形态研究，测量差异基因表达的微阵列分析，ELISA和RT-PCR，以鉴定成骨细胞和破骨细胞分化的标记，通过H-1 NMR Spectrypopicy和MAPK的定量，TWSG1和NF-- NF---NF--- KB信号通路。使用这套测定套件，我们将确定许多代谢物和生物标志物的相似性和差异，用于模拟微重力和ISS微重力环境。该项目将有助于确定可以解决的机制和途径，以减轻地球上的骨质流失。