Experimental Resources for Studies of Tenocyte Differentiation and Cell Fate Diversity

项目总结/摘要 肌腱连接肌肉和骨骼，并传递肌肉收缩产生的力。因为 由于其独特的位置，钢筋束受到显著的剪切力和压缩力， 经常肌腱损伤和撕裂。肌腱也对退行性疾病的发展具有高度抵抗性。 由于肌腱和韧带的状况以及缓慢和有限的愈合能力， 超过50%的骨科诊所就诊，给个人和社会带来了重大负担。一 近年来，肌腱和韧带状况的相对增加导致了 与肌腱相关的研究和研究赠款的数量。这项研究主要集中在 肌腱的临床应用，最近在NIAMS和研究中得到了认可 社区认为肌腱的基础研究仍然太有限，缺乏基础研究。 信息资源是肌腱研究进展的主要障碍。了这一理解 事实上，在几年前由IAMS主办的肌腱相关会议的结论中， 前然而，由于NIAMS的内部政策，RFA或NIH没有遵循这一认识 推动研究联盟，以满足这些实验的需要。 我们的实验室一直处于识别新肌腱基因和开发研究工具的最前沿， 在老鼠的肌腱研究和分享他们与肌腱社区。因此，我们决定 提出一个项目的重点和实验资源的开发和快速共享 这些资源。因此，该项目的第一个目标是将RNAseq用于各种发展和 成熟阶段，并通过比较确定肌腱分化阶段和发展为每个这些 在肌腱中具有独特表达的基因的全面和权威的列表。此外 我们将定义其表达定义这些阶段的基因的最小原型，并且可以用作 将干/祖细胞编程为肌腱细胞命运阶段的成功标准。在第二 我们将进行单细胞RNAseq转录组测定，以确定潜在的多样性， 肌腱中的亚特化肌腱细胞。在这个目标中，我们将关注的一种细胞类型是外Tenon，一种关键的 肌腱的一部分，收到非常有限的研究关注日期。最后，第三个具体目标是 为肌腱研究开发新的小鼠模型，包括额外的cre系， 肌腱细胞和相关细胞类型的靶向活性。为了培育更多的老鼠突变体， 确定调节肌腱细胞命运的基因。由于该项目被定义为资源开发 格兰特，我们将进一步专注于所有这些数据资源和鼠标线的快速传播， 支持肌腱和韧带研究的更快进展。

Project Summary/Abstract Tendons connect muscles to bones and transmit the force generated by muscle contraction. Because of their unique position, the tendons are subject to significant shear and compression forces resulting in frequent tendon damage and tears. Tendons are also highly susceptive to development of degenerative conditions and because of slow and limited healing capacity tendon and ligament conditions account to more than 50% of visits to orthopedic clinics and present a major burden to individuals and society. A relative increase in tendon and ligament conditions in recent years led to a significant increase in the number of studies and research grants associated with tendons. This research is largely focused on clinical applications in tendons and it was recently recognized at NIAMS and within the research community that the basic research in tendons remained too limited and the absence of basic information resources is a major impediment to progress in tendon research. This understanding was indeed emphasized in the conclusions of a Tendon related conference sponsored by IAMS a few years ago. However, because of internal NIAMS policies, this realization was not followed by an RFA or NIH driven research consortium to address these experimental needs. Our labs has been at the forefront of identifying new tendon genes and developing research tools for tendon research in mouse and sharing them with the tendon community. We therefore decided to propose a project focused and the development of experimental resources and the rapid sharing of these resorces. The first aim in this project is therefore to use RNAseq in various developmental and mature stages and by comparison identify tendon differentiation stages and develop for each of these stages a comprehensive and authoritative list of genes with distinctive expression in tendons. In addition we will define a prorotypic short least of genes whose expression define these stages and can be used as success criteria for the programing of stem/progenitor cells to the tendon cell fate stages. In the second specific aim we will perform single cell RNAseq transcriptome assays to identify potential diversity of sub-specialized tenocytes in tendons. One cell type we will focus on in this aim is the epitenon, a critical part of the tendons that receives very limited research attention to date. Finally, the 3rd specific aim is to develop new mouse models for tendon research including additional cre lines with more specific targeted activity in tenocytes and associated cell types. And to develop additional mouse mutants to identify genes regulating the tendon cell fate. Since this project is defined as a resource development grant we will further focus on the rapid dissemination of all these data resources and mouse lines to support a more rapid progress in tendon and ligament research.