Skeletal Phenotyping of KOMP Mice

描述(申请人提供)：骨骼疾病有一个复杂的遗传机制，反映了多个基因的总和效应，这些基因进一步被激素和个人环境改变。反映遗传和环境因素相互作用的小鼠模型将在解释人类遗传学研究方面发挥主要作用，因为小鼠的遗传背景可以得到严格控制。一个主要的例子是基因敲除表型程序(KOMP)。它的目的是对KO小鼠进行系统检查，但拟议的骨骼评估对骨骼社区没有帮助，因为它仅限于全身数字X光和DEXA。这笔赠款将表明它 在正在进行的KOMP计划中附加一个健壮的骨骼评估计划在财务和操作上都是可行的。已经建立了一个由三个地点组成的多调查员工作流程。缅因州杰克逊实验室的阿克特-比克内尔博士将接受来自KO品系的繁育小鼠，这些小鼠已经在该地点进行了动画制作、繁殖、基因分型和检查。她将注射矿化染料，并采集脊椎、四肢和头骨，运往康涅狄格州大学法明顿分校。在康涅狄格州大学，亚当斯博士将对股骨远端、椎体和头骨进行高度自动化的显微CT，以识别骨骼结构异常的KO线。罗博士的工作人员将使用计算机自动冷冻组织学方法对选定的KO系进行评估，以进行骨小梁、皮质骨和软骨生长板的动态和细胞组织形态计量，该方法已适用于矿化组织。一旦完成组织学检查的数字文件集，它就被移植到康涅狄格州大学斯托尔斯分校的Shin博士的团队，在那里，一个定制的图像分析程序测量荧光信号，以骨生物学家熟悉的术语计算组织形态计量学。当分析完成后，结果、基础图像和相关的背景表型分析将被上传到一个可联网的数据库，以便骨骼生物界可以获得结果。已经建立了对向公众提供的信息进行外部同行审查的机制。随着数据库的增长，它将成为一种宝贵的资源，使研究人员能够查询生理表型，并检索所有已定位到动态和细胞组织学水平上的查询过程中的KO基因。聚集的团队有多学科和多地点互动的记录，以协调日常活动和对KO数据集的解释。由于先前在开发高度自动化的显微CT和组织学/图像分析程序方面的投资，以及直接从KOMP生产线使用KO小鼠的能力，如果在特别的基础上进行筛查，那么进行筛查的成本可能是成本的五分之一。这是一个一次性的机会，可以利用NIH在KOMP计划中的现有投资来产生对个性化骨骼医学具有坚实的表型基础至关重要的信息。

DESCRIPTION (provided by applicant): Diseases of the skeleton have a complex genetic mechanism that reflects the summated effect of multiple genes that are further modified by hormones and the environment of the individual. Murine models that reflect the interaction of genetic and environmental factors will play a major role in interpreting human genetic studies because the genetic background of the mouse can be tightly controlled. One major example is the gene knock out phenotyping program (KOMP). It is designed to perform a systemic examination of KO mice but the proposed skeletal assessment will not be helpful to the bone community because it is limited to a full body digital X-ray and DEXA. This grant will show that it is financially and operationally feasible to append a robust skeletal evaluation program to the ongoing KOMP program. A three-site multi-investigator workflow has been established. Dr. Ackert-Bicknell at The Jackson Laboratory in Maine will receive breeder mice from the KO lines that have been animated, breed, genotyped and examined at that site. She will inject mineralization dyes and harvest the spine, limbs and skull for shipment to UCONN Farmington. At UCONN, Dr. Adams will perform a highly automated microCT of the distal femur, vertebral body and skull to identify KO lines with abnormal skeletal architecture. The selected KO lines will be assessed by Dr. Rowe's staff for dynamic and cellular histomorphometry of trabecular, cortical bone, and cartilage growth plate utilizing a computer automated cryohistological approach that has been adapted for mineralized tissues. Once the digital file set for the histological examination is completed, it is ported the Dr. Shin's group at UCONN Storrs where a custom image analysis program measures the fluorescent signals to compute the histomorphometry in terms that are familiar to the bone biologist. When the analysis is complete, the results, underlying images and relevant background phenotyping will be uploaded to a web-enabled database so that the result is available to the skeletal biology community. A mechanism for external peer review of information to be presented to the public has been established. As the database grows, it will become an invaluable resource by allowing an investigator to query for a physiological phenotype and retrieve all the KO genes that have been mapped to query process at the level of dynamic and cellular histology. The assembled teams have a track record for multidisciplinary and multisite interaction to coordinate daily activities and interpretation of KO data sets. Because of the prior investment in developing the highly automated microCT and histological/image analysis procedures, and the ability to use KO mice directly from a KOMP production line, the cost to perform the screen is probably 1/5 of the cost if performed on an ad hoc basis. This is a one-time opportunity to leverage the existing investment of NIH in the KOMP program to produce information that will be essential for personalized skeletal medicine to have a solid phenotypic foundation.