Conditional mouse models with dominant negative Osteogenesis Imperfecta

显性负性成骨不全的条件小鼠模型

基本信息

批准号：
10646852
负责人：
Matthew L Warman
金额：
$ 23.36万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10646852
关键词：
Adult Affect Age Alleles Animals Bone Density Breeding COL1A1 gene COL1A2 gene Cardiac Cell Therapy Cells Chest Chondrocytes Clinical Collagen Collagen Gene Collagen Type I Compensation Cre driver Data Disease Dominant-Negative Mutation Enterobacteria phage P1 Cre recombinase Epiphysial cartilage Evaluation Excision Foundations Funding Genes Genetic Genetic Recombination Goals Growth Heart Human Individual Introns Knowledge Laboratories Ligaments Lung Marrow Mediating Messenger RNA Modeling Mouse Strains Multiple Fractures Mus Mutation Osteoblasts Osteocytes Osteogenesis Imperfecta Patients Pharmaceutical Preparations Phenotype Production Property Proteins Publishing Research Personnel Resources Reverse Transcriptase Polymerase Chain Reaction Scientist Severities Skin Source System TNFSF5 gene Techniques Technology Tendon structure Testing The Jackson Laboratory Tissue-Specific Gene Expression Tissues United States National Institutes of Health Vertebral column body system bone bone mass cell type experience inducible Cre insight interest mouse model mutant negative affect osteoprogenitor cell prevent progenitor public repository pup radiological imaging repository reproductive rib bone structure skeletal transdifferentiation

项目摘要

The goal of this R21 application is to convert existing mouse models of moderate and severe Osteogenesis Imperfecta (OI) caused by type 1 collagen mutations to conditional mouse models of moderate and severe OI. Conditional mouse models offer several advantages over the existing strains. First, conditional mouse strains can be maintained and distributed by public repositories; existing strains are in individual labs, and not in repositories, because of their skeletal fragility. Second, OI is a multi-system disease that affects bone, heart, lung, tendon, ligament, and skin. When studying OI, the phenotype in one organ system (e.g., heart or lung) is confounded by disease-associated problems in another organ system (e.g., spine and ribs). Conditional mouse models in which Cre-recombinase is used to express the mutant protein in a tissue-specific manner minimizes this confounding. Currently, there are no conditional mouse models with type 1 collagen mutations, even though ~ 85% of human OI is caused by such mutations. Third, by converting existing strains to conditional strains, any new data obtained with the conditional strains can be compared to previously published data because the same mutation was studied. We will modify existing alleles by inserting a gene-trap using i- GONAD gene-editing technology. Applying gene-traps and i-GONAD will enable us to efficiently convert 4 existing OI-causing alleles (Aga2, Jrt, Brtl, and p.G610C) in mice to conditional OI-causing alleles. Mouse strains with the new conditional alleles can be distributed by public repositories. We will also determine the effect of inducing endogenous expression of mutant type 1 collagen in hypertrophic chondrocytes, since hypertrophic chondrocytes have been shown to transdifferentiate into osteoblasts and osteoprogenitors. Therefore, it is important to know if bone properties are strongly influenced by cells expressing mutant type 1 collagen that have descended from hypertrophic chondrocytes, or if unaffected bone forming cells descended from other progenitor sources compensate for, or outcompete, the mutant hypertrophic chondrocyte-derived descendants. This latter knowledge will aid investigators who are interested in using cell therapies to displace/replace mutant osteoblasts and osteoprogenitors with wild-type ones.

该 R21 应用程序的目标是转换现有的中度和重度成骨的小鼠模型中度和重度 OI 条件小鼠模型由 1 型胶原蛋白突变引起的不完美 (OI)。与现有品系相比，条件小鼠模型具有多种优势。一、条件小鼠品系可以由公共存储库维护和分发；现有菌株位于各个实验室，而不是在储存库，因为它们的骨骼脆弱。其次，成骨不全症是一种多系统疾病，影响骨骼、心脏、肺、肌腱、韧带和皮肤。研究 OI 时，一个器官系统（例如心脏或肺）的表型是被另一个器官系统（例如脊柱和肋骨）的疾病相关问题所困扰。有条件的鼠标使用 Cre 重组酶以组织特异性方式表达突变蛋白的模型最小化这种混淆。目前，还没有条件性的1型胶原蛋白突变小鼠模型，甚至尽管约 85% 的人类成骨不全症是由此类突变引起的。第三，通过将现有菌株转化为有条件菌株菌株，使用条件菌株获得的任何新数据都可以与之前发布的数据进行比较因为研究了相同的突变。我们将通过使用 i- 插入基因陷阱来修改现有的等位基因性腺基因编辑技术。应用基因陷阱和 i-GONAD 将使我们能够有效地转化 4 将小鼠中现有的导致 OI 的等位基因（Aga2、Jrt、Brtl 和 p.G610C）转换为条件性 OI 导致的等位基因。老鼠具有新条件等位基因的菌株可以通过公共存储库分发。我们还将确定在肥大软骨细胞中诱导突变型 1 型胶原内源表达的效果，因为肥大软骨细胞已被证明可以转分化为成骨细胞和骨祖细胞。因此，了解表达突变型 1 的细胞是否强烈影响骨骼特性非常重要来自肥大软骨细胞的胶原蛋白，或者未受影响的骨形成细胞的胶原蛋白来自其他祖细胞来源的细胞补偿或战胜突变的肥大软骨细胞衍生的后代。后者的知识将帮助有兴趣使用细胞疗法的研究人员用野生型取代/替换突变的成骨细胞和骨祖细胞。