Skeletal Phenotyping of KOMP Mice

描述（申请人提供）：骨骼疾病具有复杂的遗传机制，反映了多个基因的综合效应，这些基因进一步受到激素和个体环境的修饰。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学研究中发挥重要作用，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分析程序（KOMP）。它旨在对 KO 小鼠进行系统检查，但所提出的骨骼评估对骨骼界没有帮助，因为它仅限于全身数字 X 射线和 DEXA。这笔赠款将表明 在正在进行的 KOMP 计划中添加一个强大的骨骼评估计划在财务和操作上都是可行的。 已经建立了三站点多研究者工作流程。缅因州杰克逊实验室的 Ackert-Bicknell 博士将接收来自 KO 品系的种鼠，这些小鼠已在该地点进行了活化、繁殖、基因分型和检查。她将注射矿化染料并采集脊柱、四肢和头骨，然后运往康涅狄格大学法明顿分校。在康涅狄格大学，Adams 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 系。 Rowe 博士的工作人员将利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 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。一旦用于组织学检查的数字文件集完成，它就会被移植到 UCONN Storrs 的 Shin 博士小组，其中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，以便骨骼生物学界可以使用结果。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，允许研究人员查询生理表型并检索已映射到动态和细胞组织学水平的查询过程的所有 KO 基因。组建的团队拥有多学科和多地点互动的记录，可以协调日常活动和 KO 数据集的解释。 由于先前投资开发高度自动化的 microCT 和组织学/图像分析程序，并且能够直接使用来自 KOMP 生产线的 KO 小鼠，因此执行筛选的成本可能是临时执行成本的 1/5。这是一个一次性的机会，可以利用 NIH 在 KOMP 项目中的现有投资来产生对于个性化骨骼医学奠定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.