Mechanisms of growth plate patterning revealed by natural variation in mammalian ossification

哺乳动物骨化自然变异揭示生长板图案形成机制

• 批准号: 1656315
• 负责人: Philip Reno
• 金额: $ 80万
• 依托单位: Philadelphia College of Osteopathic Medicine
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1656315&HistoricalAwards=false
• 关键词: Mechanisms; growth; plate; patterning; revealed

Skeletal growth occurs at specific locations called growth plates. These preserve pools of cartilage cells, or chondrocytes, that enable rapid bone growth. Much is known regarding the genes and developmental processes that control chondrocyte differentiation and proliferation. Yet, how growth plates are induced and why they occur at specific locations in the skeleton continues to be a mystery. For example, metatarsal bones in the feet produce a growth plate at only one end, and the pisiform and calcaneus are the only wrist and ankle bones to form a growth plate. By comparing these growth plates to the matched skeletal sites that fail to form them, these bones form ideal models to isolate the patterns of gene expression and regulation that control how and where growth plates are formed. The pisiform growth plate is particularly susceptible to the perturbation of key developmental genes that further allows the role of these genes in growth plate formation to be identified. This program expands laboratory research opportunities for a diverse body of undergraduate and graduate students in the application of cutting-edge methods in developmental biology and comparative genomics to fundamental questions related to human development and growth. This research project supports collaboration between scientists, students, and the College of Physicians of Philadelphia to develop outreach efforts to underserved populations lacking in such opportunities. A science workshop for gifted but underserved Philadelphia high school students will provide classroom and research experiences related to mammalian developmental biology and gene regulation.This research will employ a novel approach comparing otherwise equivalent regions of the endochondral skeleton that do and do not form growth plates. Through whole transcriptome (RNA-seq) and chromatin-state (ATAC-seq) sequencing of paired growth plate forming and non-forming tissues, the pisiform, calcaneus, and metatarsal will identify inductive factors of growth plate formation. It is hypothesized that expression differences exist in chondrocyte populations prior to the visible formation of growth plates and that these factors will also be differentially associated with reserve zone and perichondrial cells. In addition, Hox genes have a role in growth plate patterning, and it is believed that the nodular nature of most carpals and tarsals results from the more limited role for Hox genes in their development. However, Hoxa11 and Hoxd11 loss-of-function result in malformation of the proximal carpus and tarsus, including shortened pisiforms. Thus, the pisiform and calcaneus are natural test cases to determine the role that later stage Hox11 expression plays in growth plate formation. It is predicted that Hox11 has a direct role in pisiform and calcaneus growth plate induction by establishing reserve zone and perichondrial chondrocyte populations. Furthermore, it is predicted that growth plate forming sites will be biased for active enhancers containing Hox11-13 response elements and that enhancer activity patterns will be shared among different growth plates.

骨骼生长发生在称为生长板的特定位置。这些细胞保存了软骨细胞池，使骨骼能够快速生长。关于控制软骨细胞分化和增殖的基因和发育过程已经知道很多。然而，生长板是如何被诱导的，以及为什么它们会出现在骨骼的特定位置仍然是一个谜。例如，足部的跖骨仅在一端产生生长板，豌豆骨和跟骨是形成生长板的唯一手腕和脚踝骨。通过将这些生长板与未能形成它们的匹配骨骼部位进行比较，这些骨骼形成了理想的模型，以分离控制生长板形成方式和位置的基因表达和调控模式。豌豆状生长板特别容易受到关键发育基因的干扰，这进一步允许鉴定这些基因在生长板形成中的作用。该计划为本科生和研究生的多元化机构扩大了实验室研究机会，将发展生物学和比较基因组学的前沿方法应用于与人类发育和生长有关的基本问题。该研究项目支持科学家，学生和费城医师学院之间的合作，为缺乏此类机会的服务不足人群开展外展工作。一个科学研讨会的天才，但缺乏服务的费城高中学生将提供课堂和研究经验，有关哺乳动物发育生物学和基因regulation.This研究将采用一种新的方法，比较否则等同区域的软骨内骨骼，做和不形成生长板。通过成对生长板形成和非形成组织的全转录组（RNA-seq）和染色质状态（ATAC-seq）测序，豌豆骨、跟骨和跖骨将鉴定生长板形成的诱导因子。据推测，在可见的生长板形成之前，软骨细胞群中存在表达差异，并且这些因子也将与储备区和软骨膜细胞差异相关。此外，Hox基因在生长板模式中具有作用，并且据信大多数腕骨和跗骨的结节性质是由于Hox基因在其发育中的作用更有限。然而，Hoxa 11和Hoxd 11功能丧失导致近端腕骨和跗骨畸形，包括豌豆骨缩短。因此，豌豆骨和跟骨是确定后期Hox 11表达在生长板形成中所起作用的天然测试案例。据预测，Hox 11有一个直接的作用，豌豆骨和跟骨生长板诱导建立储备区和软骨膜软骨细胞群体。此外，据预测，生长板形成网站将偏向于含有Hox 11 -13响应元件的活性增强子，并且增强子活性模式将在不同的生长板之间共享。

项目成果

Ossification pattern of the unusual pisiform in two‐toed ( Choloepus ) and three‐toed sloths ( Bradypus )

二趾树懒 (Choloepus) 和三趾树懒 (Bradypus) 中不寻常的豌豆骨的骨化模式

• DOI: 10.1002/ar.24832
• 发表时间: 2021
• 期刊: The Anatomical Record
• 作者: Gavazzi, Lia M.;Kjosness, Kelsey M.;Reno, Philip L.
• 通讯作者: Reno, Philip L.

Identifying growth plate specific gene using the unusual ossification of the mammalian metatarsal and pisiform

利用哺乳动物跖骨和豌豆骨的异常骨化识别生长板特异性基因

• 发表时间: 2023
• 期刊: American journal of biological anthropology
• 作者: Reno Philip L.;Kjosness, Kelsey M.;Park, Sundae;Wilson, Lauren;Doelp, Sarah N.;Wallace, Sherrie A.;Menke, Douglas B.
• 通讯作者: Menke, Douglas B.

Influence of Hoxa11 and Hoxd11 on Calcaneus Growth and Ossification

Hoxa11和Hoxd11对跟骨生长和骨化的影响

• 发表时间: 2021
• 期刊: The FASEB journal
• 作者: Bhavsar, Avani;Goldman, Samuel;Kjosness, Kelsey M;Schuetz, Emily;Kistler, Sabrina;Thompson, Ryan;Reno, Philip L
• 通讯作者: Reno, Philip L

Differential Impact of Hoxa11 and Hoxd11 on Pisiform Ossification and Epiphysis Formation

Hoxa11 和 Hoxd11 对豌豆骨骨化和骨骺形成的不同影响

• 发表时间: 2021
• 期刊: The FASEB journal
• 作者: Graziano, Timothy;Sweeney, Brendan;Kjosness, Kelsey M;Kistler, Sabrina;Schuetz, Emily;Thompson, Ryan;Reno, Philip
• 通讯作者: Reno, Philip

Morphological Changes in Calcanei of Mice with Hoxa11 and Hoxd11 loss-of-function mutations

Hoxa11 和 Hoxd11 功能丧失突变小鼠跟骨的形态变化

• 发表时间: 2021
• 期刊: The FASEB journal
• 作者: Goldman, Samuel;Bhavsar, Avani;Kjosness, Kelsey M;Kistler, Sabrina;Schuetz, Emily;Stephens, Nicholas;Ryan, Timothy M;Reno, Philip L
• 通讯作者: Reno, Philip L