Skeletal Phenotyping of KOMP Mice

描述（由申请人提供）：骨骼疾病具有复杂的遗传机制，反映了多个基因的总和效应，这些基因被个体的激素和环境进一步修饰。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研究中发挥重要作用，因为小鼠的遗传背景可以被严格控制。一个主要的例子是基因敲除表型程序（KOMP）。其设计用于对KO小鼠进行全身检查，但拟定的骨骼评估对骨群落没有帮助，因为其仅限于全身数字X射线和DEXA。这份补助金将表明， 在财务上和操作上可行，可以在正在进行的KOMP项目中附加一个强大的骨骼评估项目。 已经建立了三个研究中心多名研究者的工作流程。位于缅因州的杰克逊实验室的阿克特-比克内尔博士将接收来自KO品系的育种小鼠，这些小鼠在该地点进行了动画、繁殖、基因分型和检查。她将注射矿化染料，并收获脊柱，四肢和头骨运往康州大学法明顿。在UCONN，亚当斯博士将对股骨远端、椎体和颅骨进行高度自动化的microCT，以识别具有异常骨骼结构的KO线。由Rowe博士的工作人员使用适用于矿化组织的计算机自动冷冻组织学方法，对选定的KO细胞系进行小梁、皮质骨和软骨生长板的动态和细胞组织形态计量学评估。一旦完成组织学检查的数字文件集，它就被移植到UCONN斯托尔斯的Shin博士小组，在那里，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。分析完成后，结果、基础图像和相关背景表型将上传到网络数据库，以便骨骼生物学社区可以获得结果。已经建立了一个机制，对向公众提供的信息进行外部同行审查。随着数据库的增长，它将成为一个宝贵的资源，允许研究人员查询生理表型和检索所有的KO基因，已映射到查询过程中的动态和细胞组织学水平。集合的团队有多学科和多地点互动的跟踪记录，以协调日常活动和KO数据集的解释。 由于在开发高度自动化的microCT和组织学/图像分析程序方面的先前投资，以及直接使用来自KOMP生产线的KO小鼠的能力，执行筛选的成本可能是如果临时执行的成本的1/5。这是一个一次性的机会，利用NIH在KOMP计划中的现有投资来产生信息，这些信息对于个性化骨骼医学具有坚实的表型基础至关重要。

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.