Regeneration of Nucleus Pulposus by Tissue Engineering

组织工程髓核再生

• 批准号: 8468642
• 负责人: Makarand V Risbud
• 金额: $ 31.64万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-02-01 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8468642
• 关键词: Address; Adult; Animal Model; Back Pain; Biological; Cell Differentiation process; Cell Proliferation; Cells; Cellularity; Development; Disease; Etiology; Funding; Future; Gene Targeting; Genes; Genetic Engineering; Goals; Health; Human; Hypoxia; In Vitro; Intervertebral disc structure; Investigation; Knockout Mice; Learning; Ligands; Link; Low Back Pain; Maintenance; Measures; Mechanics; Molecular Genetics; Morbidity - disease rate; Mus; Natural regeneration; Oryctolagus cuniculus; Pathology; Patients; Phenotype; Population; Rattus; Regulation; Role; Signal Transduction; Societies; Source; Spatial Distribution; Stem cells; Subfamily lentivirinae; System; Techniques; Testing; Time; Tissue Engineering; Tissues; Vertebral column; authority; cell type; functional restoration; gain of function; in vivo; insight; intervertebral disk degeneration; loss of function; notch protein; notochord; novel strategies; nucleus pulposus; progenitor; promoter; repaired; selective expression; tissue repair

DESCRIPTION (provided by applicant): The goal of the competitive renewal is to test the hypothesis that in the hypoxic niche of the intervertebral disc, notch signaling is required for maintenance of proliferation and differentiation of cell populations. The objective of the first Specific Aim is to characterize the expression of notch and notch ligands and to investigate the importance of notch signaling in proliferation and differentiation of disc cells; to assess notch and notch ligand expression in tissue isolated from patients with degenerative disc disease. Using loss and gain of function studies we will determine the importance of the notch system in terms of cell proliferation and acquisition of the mature phenotype. By evaluating the localization of notch receptors and ligands we will gain insight into the interaction between different cell types within the disc. The goal of the second Specific Aim is to ascertain if in the hypoxic microenvironment notch and HIF signaling co-regulate expression of selective notch receptors, ligands and notch target genes in disc cells; in addition, we will determine if notch target genes Hey1 and Hey2 influence HIF function. In parallel, the interaction between notch and HIF will be assessed to evaluate the involvement of notch-ICD in the regulation of HIF-1 target genes. A final goal is to examine proliferative cells isolated from degenerate human discs for the expression and activities of notch signaling components. In the third Aim we will use a notochord specific knockout mice to investigate the importance of notch signaling in governing cell proliferation, survival and function during disc development and to delineate the role of notch in modulating HIF-1 function in the disc. Since we will also delete HIF-1, these studies will reveal the importance of this gene in regulating notch activity. The goal of the last Specific Aim is to develop an animal model to learn if inactivation of notch activity promotes a loss of cellularity, a decrease in number of differentiated cells and eventual degeneration of the intervertebral disc. We will inject rabbit intervertebral discs with lentivirus encoding psen1/psen2-SiRNA to suppress expression of endogenous notch activity. We will measure the activities of components of the notch signaling system, HIF function, the activities and numbers of proliferative cells and assess the degenerative status of the disc itself. These studies will validate and further extend the investigations described above, and provide new information on the critical role of notch signaling in terms of proliferation, survival, and differentiation of disc cells. Results of the investigation should provide a new understanding of how cell proliferation is regulated in the disc and provide a strategy for future use of pharmacological agents, and molecular genetic or tissue engineering approaches for effecting tissue repair utilizing endogenous disc cells.

描述（由申请人提供）：竞争性更新的目的是检验以下假设：在椎间盘的低氧小境中，保持缺口信号来维持增殖和细胞种群的分化。第一个特定目的的目的是表征缺口和缺口配体的表达，并研究Notch信号在盘细胞增殖和分化中的重要性。评估从退行性椎间盘突出的患者中分离的组织中的Notch和Notch配体表达。使用功能研究的丧失和增益，我们将根据细胞增殖和成熟表型的获取来确定Notch系统的重要性。通过评估Notch受体和配体的定位，我们将深入了解椎间盘中不同细胞类型之间的相互作用。第二个特定目的的目的是确定在低氧微环境缺口和HIF信号传导中是否共同调节了椎间盘细胞中选择性缺口受体，配体和凹口靶基因的表达；此外，我们将确定Notch靶基因HEY1和HEY2是否影响HIF功能。同时，将评估Notch和HIF之间的相互作用，以评估Notch-ICD参与HIF-1靶基因的调节。最终目标是检查从退化的人盘分离出的增生细胞，以表达和活性。在第三个目的中，我们将使用脊索特定的敲除小鼠来研究Notch信号在椎间盘发育过程中控制细胞增殖，存活和功能中的重要性，并描述Notch在调节光盘中HIF-1功能中的作用。由于我们还将删除HIF-1，因此这些研究将揭示该基因在调节Notch活性中的重要性。最后一个特定目的的目的是开发动物模型，以了解Notch活性的失活是否促进了细胞的丧失，分化细胞数量的减少以及椎间盘的最终变性。我们将以编码PSEN1/PSEN2-SIRNA抑制内源性缺口活性的慢病毒注入兔椎间盘。我们将测量Notch信号系统，HIF功能，增殖细胞的活性和数量的组件活性，并评估光盘本身的退化状态。这些研究将验证并进一步扩展上述研究，并提供有关Notch信号在椎间盘细胞增殖，存活和分化方面的关键作用的新信息。研究的结果应提供对椎间盘中细胞增殖如何受到调节的新理解，并为未来使用药理学剂以及分子遗传或组织工程方法提供了利用内源性盘细胞的组织修复的策略。

项目成果

Stem cells in craniofacial and dental tissue engineering.

• DOI: 10.1111/j.1601-6343.2005.00324.x
• 发表时间: 2005-05
• 期刊: Orthodontics & craniofacial research
• 影响因子: 3.1
• 作者: MV Risbud;IM Shapiro

Reply.

回复。

• DOI: 10.1002/art.40923
• 发表时间: 2019
• 期刊: Arthritis & rheumatology (Hoboken, N.J.)
• 作者: Kim,AlfredHJ;Strand,Vibeke;Atkinson,JohnP
• 通讯作者: Atkinson,JohnP

An organ culture system to model early degenerative changes of the intervertebral disc.

• DOI: 10.1186/ar3494
• 发表时间: 2011
• 期刊: Arthritis research & therapy
• 影响因子: 4.9
• 作者: Ponnappan RK;Markova DZ;Antonio PJ;Murray HB;Vaccaro AR;Shapiro IM;Anderson DG;Albert TJ;Risbud MV
• 通讯作者: Risbud MV