Testing the tissue-specific mechanisms of Hoxa5 function in musculoskeletal patterning

测试 Hoxa5 在肌肉骨骼模式中功能的组织特异性机制

• 批准号: 9351838
• 负责人: Jennifer H Mansfield
• 金额: $ 38.95万
• 依托单位: BARNARD COLLEGE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-09 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9351838
• 关键词: Address; Alleles; Animals; Anterior; Arts; Behavior; Binding Sites; Body Patterning; Bone Development; Cartilage; Cell physiology; Cells; Cervical; Chest; Chondrocytes; Chromatin Structure; Congenital Abnormality; Connective Tissue; Connective Tissue Diseases; Dermis; Development; Disease; Dissection; Dissociation; Elements; Embryo; Endothelium; Epitopes; Flow Cytometry; Gene Targeting; Genes; Genetic; Genetic Transcription; Genomics; Genotype; Goals; HOX protein; Health; High-Throughput Nucleotide Sequencing; Homeobox Genes; Human; Label; Lead; Ligaments; Light; Maintenance; Malignant Neoplasms; Mammals; Maps; Mediating; Molecular; Molecular Genetics; Morphogenesis; Morphology; Mus; Muscle; Muscle Development; Musculoskeletal; Musculoskeletal Development; Mutation; Osteogenesis; Other Genetics; Pathway interactions; Pattern; Phenotype; Play; Positioning Attribute; Process; Proteins; Regulation; Research; Resources; Role; Sclerotome; Signal Transduction; Skeleton; Somites; Specific qualifier value; Specificity; Students; Tendon structure; Testing; Tissues; Training; Underrepresented Groups; Variant; Vertebral column; Vertebrates; Woman; Work; base; bone; cartilage development; chromatin immunoprecipitation; cofactor; college; design; differential expression; experimental study; genetic approach; in vivo; laboratory curriculum; ligament development; loss of function; polyclonal antibody; progressive myositis ossificans; rib bone structure; skeletal; skeletal tissue; student participation; tendon development; transcription factor; transcriptome sequencing; undergraduate student

Abstract Development of cartilage, bones, muscles, tendons and ligaments must be highly coordinated, with tissue type differentiation and morphogenesis occuring in a concerted way that allows the resulting tissues to function together. The axial musculoskeleton (vertebral column and ribs) develops from the somites, in which all musculoskeletal tissue types are specified and develop coordinately. However, the morphology of somite derivatives varies with position along the body axis. This variation is controlled by Hox proteins, conserved transcription factors that pattern the body axis of most animal embryos. In vertebrates, Hox proteins both confer anterior-posterior identity on nascent segments and play direct roles in tissue morphogenesis later in development. However, we still know relatively little about their mechanisms of action, including the cell and tissue types in which they act, the cellular behaviors they regulate, and ultimately their transcriptional targets. Hoxa5 non-redundantly patterns musculoskeletal elements at the cervical-thoracic transition. In order to understand how it acts, we used genetic lineage labeling to fate map the descendants of Hoxa5 expressing cells. We found that Hoxa5 descendants contribute to a restricted number tissue types, such as cartilage and perichondrium, but only contribute rarely to tendon and never to muscle. This restriction of Hoxa5 descendant fate may reflect an important aspect of its function. Here, we propose to investigate the mechanism through which Hoxa5 patterns the cervical-thoracic transition musculoskeleton of mice, using a combination of genetic lineage labeling, conditional and loss-of-function analysis, and high throughput sequencing. Our specific aims are designed to: (1) Identify the tissue specificity of Hoxa5 action in patterning the axial skeleton (2) Identify genes and gene networks regulated cell-autonomously by Hoxa5 and (3) Identify direct transcriptional targets of Hoxa5 in the somites. Successful completion of this project will shed light on the mechanisms through which axial patterning is regulated by Hox genes, and can also be applied to a general understanding the mechanisms of Hoxa5 in other tissue types and cancers. More generally, the work is relevant to human health through its application to understanding musculoskeletal tissue patterning and differentiation during normal development and disease. Finally, this work will be conducted with undergraduates at Barnard College, a liberal arts college for women. It will be performed by research students in the PI’s lab, and will be introduced into a newly developed, full year laboratory course at Barnard. Course-based research has been shown to increase participation of students, including those from underrepresented groups, in STEM and to increase the pursuit of postgraduate STEM training. This course will engage undergraduates in substantive original research while providing them with training in current molecular genetics approaches.

抽象的 软骨、骨骼、肌肉、肌腱和韧带的发育必须高度协调， 组织类型分化和形态发生以协调一致的方式发生，从而允许产生 共同发挥功能的组织。轴向肌肉骨骼（脊柱和肋骨）由 体节，其中所有肌肉骨骼组织类型均被指定并协调发育。 体节衍生物的形态随体轴位置的变化而变化。 Hox 蛋白是保守的转录因子，形成大多数动物胚胎的体轴。 在脊椎动物中，Hox 蛋白既赋予新生节段的前后身份，又直接发挥作用 然而，我们对它们在发育后期组织形态发生中的作用仍然知之甚少。 作用机制，包括它们起作用的细胞和组织类型、它们的细胞行为 调节，并最终它们的转录目标非冗余地模式肌肉骨骼。 为了了解其作用方式，我们使用了遗传谱系。 标记 Hoxa5 表达细胞的后代的命运图谱 我们发现 Hoxa5 后代。 有助于有限数量的组织类型，例如软骨和软骨膜，但仅有助于 Hoxa5 后代命运的这种限制可能反映了一个重要的因素。 在这里，我们建议研究 Hoxa5 模式化的机制。 小鼠颈胸过渡肌骨骼，使用遗传谱系标记的组合， 我们的具体目标是条件分析和功能丧失分析以及高通量测序。 旨在：(1) 识别 Hoxa5 在形成中轴骨骼模式中的组织特异性 (2) 识别 Hoxa5 细胞自主调节的基因和基因网络，以及 (3) 识别直接 该项目的成功完成将揭示 Hoxa5 在体节中的转录目标。 Hox 基因调节轴向模式的机制，也可以应用于 一般了解 Hoxa5 在其他组织类型和癌症中的机制。 工作通过其应用于了解肌肉骨骼组织而与人类健康相关 最后，这项工作将是正常发育和疾病期间的模式和分化。 它将在巴纳德学院（一所女子文理学院）的本科生中进行。 由 PI 实验室的研究生进行，并将被引入新开发的全年实验室 巴纳德的课程已被证明可以提高学生的参与度， 包括那些来自 STEM 领域代表性不足的群体，并增加对研究生的追求 本课程将让本科生参与实质性的原创研究，同时提供 STEM 培训。 他们接受了当前分子遗传学方法的培训。

项目成果

HOXA5 Participates in Brown Adipose Tissue and Epaxial Skeletal Muscle Patterning and in Brown Adipocyte Differentiation.

HOXA5 参与棕色脂肪组织和轴外骨骼肌模式以及棕色脂肪细胞分化。

• 发表时间: 2021
• 作者: Holzman, Miriam A;Ryckman, Abigail;Finkelstein, Tova M;Landry;Schindler, Kyra A;Bergmann, Jenna M;Jeannotte, Lucie;Mansfield, Jennifer H
• 通讯作者: Mansfield, Jennifer H

HOXA5 protein expression and genetic fate mapping show lineage restriction in the developing musculoskeletal system.

HOXA5 蛋白表达和遗传命运图谱显示发育中的肌肉骨骼系统中的谱系限制。

• 发表时间: 2018
• 作者: Holzman, Miriam A;Bergmann, Jenna M;Feldman, Maya;Landry;Jeannotte, Lucie;Mansfield, Jennifer H
• 通讯作者: Mansfield, Jennifer H

Hoxa5 Activity Across the Lateral Somitic Frontier Regulates Development of the Mouse Sternum.

Hoxa5 跨侧体细胞边界的活性调节小鼠胸骨的发育。

• 发表时间: 2022
• 作者: Mitchel, Kira;Bergmann, Jenna M;Brent, Ava E;Finkelstein, Tova M;Schindler, Kyra A;Holzman, Miriam A;Jeannotte, Lucie;Mansfield, Jennifer H
• 通讯作者: Mansfield, Jennifer H