Skeletal Phenotyping of Heterozygotes from IMPC Embryonic Lethal Lines

IMPC 胚胎致死系杂合子的骨骼表型

基本信息

批准号：
10622475
负责人：
PETER MAYE
金额：
$ 51.9万
依托单位：
UNIVERSITY OF CONNECTICUT SCH OF MED/DNT
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-04-02 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10622475
关键词：
Adult Affect Animals Architecture Area Big Data Bioinformatics Biological Biology Body Composition Body Size Bone Marrow Breeding Calibration Cartilage Cell Nucleus Chromosome Mapping Classification Collagen Communities Complement Critiques Cryopreservation Data Developmental Biology Diameter Dimensions Distal Dyes Embryo Embryonic Development Engineering Enzyme-Linked Immunosorbent Assay Epiphysial cartilage Face Fatty acid glycerol esters Femur Fiber Formalin Frequencies Funding Agency Genes Genotype Goals Grant Harvest Health Heterozygote Histologic Human Image Analysis Informatics Injections International Investments Knock-out Knockout Mice Knowledge Life Literature Location Maps Marrow Microscopy Minerals Modality Molecular Mus Myelogenous Osteoblasts Osteoclasts Osteogenesis Outcome Pathology Pathway interactions Pattern Peptides Phenotype Physiologic calcification Physiological Population Positioning Attribute Pregnancy Primary Cell Cultures Process Production Program Development Protocols documentation Recommendation Research Research Personnel Sampling Serum Shapes Skeletal Development Skeletal Muscle Skeleton Slide Soleus Muscle Source Spectrum Analysis Stains Structure Testing Tissues United States National Institutes of Health Variant Vertebral column Work animation bone bone mass cortical bone crosslink design expectation experience fetal insight knockout gene lean body mass mutant osteoblast differentiation osteoclastogenesis outreach phenotypic data programs repository sample collection screening skeletal skeletal abnormality spine bone structure substantia spongiosa tibia transcriptome sequencing videoconference web portal young adult

项目摘要

Abstract The International Mouse Phenotyping Consortium (IMPC) has developed an invaluable repository of gene knockout (KO) mice and has performed a whole-animal assessment of each KO line. Approximately 1/3 of the KO lines are embryonic lethals that were characterized as heterozygous (HET) KO lines. Previously we performed high throughput µCT and histomorphometric analysis on 220 unselected viable homozygous (HOM) KO lines. This study found that ~12.5% have an unequivocal variance in total bone mass (trabecular bone volume and/or bone size), most of which were not detected by the IMPC screening. The results and interpretation of the KOs examined in the screen are available on a web portal (bonebase.org). Based on this experience, the phenotyping data on the IMPC webportal and the literature, we estimate that the HET population of KOs will be an equally rich source of genes that affect bone/body variance and skeletal health in the later adult years. This proposal will apply our skeletal phenotyping workflow to selected HET KO lines that are likely to have a significant bone/body mass phenotype. Both µCT and total body composition as determined by TD-NMR will be used as the screening modality. HET KO lines with significant variance in bone architecture will be processed for histomorphometry. Based on those results, certain lines will undergo studies of osteogenesis (Raman microscopy for the mineral/matrix composition, primary cell culture for osteoblast) and/or osteoclastogenesis (collagen crosslinks and primary cell culture for osteoclast). In HET KO lines with variance in body composition, histomorphometry of the skeletal muscle will be performed. The intent is to discriminate the impact of the HET KO as acting directly on the osteoblast or osteoclast, or whether the observed phenotype is a secondary effect of the HET KO on other tissues that can influence bone. In addition, HOM KO lethal lines that survive to 18.5 days of gestation will undergo a gross and histological skeletal examination to determine the impact of the gene on skeletal development. To complement these biological findings, a bioinformatics component is being added to map the KO genes that are affecting osteoblast or osteoclast directly to known or postulated molecular pathways. Does the gene participate in the function of the network or is it a product of a network? The final goal of the project is to place the biological and bioinformatic data into a classification structure that reveals differences and similarities between the various KO lines. We will engage internal and external experts in bone biology to review the data assembled for a specific HET KO line to critique and amplify our interpretation and ever-evolving classification schema. Recognizing that frequency and complexity of the genes affecting bone is a big-data challenge (estimated to be ~3,500 genes), our intent is to lay the ground work for how this information will be gathered, presented, interpreted, queried and eventually applied toward the goal of personalized skeletal human health.

抽象的国际小鼠表型联盟（IMPC）开发了一个宝贵的基因存储库敲除（KO）小鼠，并对每条KO线进行了全面评估。大约1/3 KO线是胚胎致死性的，其特征为杂合（HET）KO系。以前我们对220个未选择的生存纯合（HOM）进行了高吞吐量µCT和组织形态分析 KO线。这项研究发现，〜12.5％的总骨质量有明确的差异（小梁骨体积和/或骨骼大小），其中大多数未通过IMPC筛选检测到。结果和屏幕上检查的KO的解释可在Web门户网站（Bonebase.org）上获得。基于此经验，关于IMPC网络竞争和文献的表型数据，我们估计 KO的种群将是影响骨/身体方差和骨骼健康的基因的同样丰富的来源后来的成年岁月。该建议将我们的骨骼表型工作流程应用于选定的het ko线可能具有明显的骨/体重表型。 µCT和总体组成由TD-NMR确定将用作筛选方式。 HET KO线的骨骼显着差异架构将用于组织形态计量学。基于这些结果，某些线将进行研究成骨（用于矿物/基质组成的拉曼显微镜，成骨细胞的原代细胞培养）和/或破骨细胞生成（胶原蛋白交联和破骨细胞的原发性细胞培养）。在Het KO线上人体组成方差，将进行骨骼肌的组织形态法。意图是区分HET KO的影响直接作用于成骨细胞或破骨细胞上，或者是观察到的表型是HET KO对其他可能影响骨骼的次要作用。此外，妊娠18.5天的HOM KO致命线将经历粗大和组织学骨骼检查基因对骨骼发育的影响。完成这些生物学调查结果，正在添加生物信息学成分以映射影响成骨细胞或直接到已知或张贴的分子途径的破骨细胞。基因参与了网络还是网络的产物？该项目的最终目标是放置生物学和生物信息分类结构的数据揭示了各种KO线之间的差异和相似性。我们将吸引骨生物学的内部和外部专家，以审查为特定HET KO组装的数据批判并扩大我们的解释和不断发展的分类模式。认识到这一点影响骨骼的基因的频率和复杂性是一个大数据挑战（估计约为3500个基因），我们的目的是为如何收集，呈现，解释，询问这些信息奠定基础工作并最终应用于个性化骨骼人类健康的目标。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Predicting the targets of IRF8 and NFATc1 during osteoclast differentiation using the machine learning method framework cTAP.

DOI：
10.1186/s12864-021-08159-z
发表时间：
2022-01-07
期刊：
BMC genomics
影响因子：
4.4
作者：
Wang H;Joshi P;Hong SH;Maye PF;Rowe DW;Shin DG
通讯作者：
Shin DG

Mesenchyme-specific loss of Dot1L histone methyltransferase leads to skeletal dysplasia phenotype in mice.

DOI：
10.1016/j.bone.2020.115677
发表时间：
2021-01
期刊：
Bone
影响因子：
4.1
作者：
Sutter PA;Karki S;Crawley I;Singh V;Bernt KM;Rowe DW;Crocker SJ;Bayarsaihan D;Guzzo RM
通讯作者：
Guzzo RM

Intrafibrillar Mineralized Collagen-Hydroxyapatite-Based Scaffolds for Bone Regeneration.