Formation of the intervertebral disk: a molecular approach

椎间盘的形成：分子方法

• 批准号: 7777107
• 负责人: BRIAN David HARFE
• 金额: $ 16.38万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2010-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7777107
• 关键词: Adult; Affect; American; Animal Model; Back Pain; Cell Therapy; Cells; Collaborations; Development; Developmental Biology; Elderly; Embryo; Florida; Future; Genetic; Goals; Healed; Injury; Intervertebral disc structure; Laboratories; LacZ Genes; Mechanics; Modeling; Molecular; Mus; Parents; Patients; Pennsylvania; Population; Proliferating; Proteins; Publishing; Role; Stem cells; Testing; Therapeutic; Tissue Engineering; Translating; United States; Universities; Work; base; cell type; cost; experience; healing; intervertebral disk degeneration; molecular marker; mouse model; notochord; nucleus pulposus; parent grant; precursor cell; research study; spine bone structure

DESCRIPTION (provided by applicant): It has been estimated that two-thirds of Americans will experience at least one episode of back pain in their lifetime. Back pain affects more than half of people over the age of 65 and the treatment of back pain costs 50- 100 billion dollars per year in the United States. Most back pain is thought to result from damage to the intervertebral disc, which are located between each vertebra. Presently there is no cure for intervertebral discs that have been damaged either by physical injury or have degenerated. The goal of the parent R01 proposal is to determine the cellular and molecular mechanism(s) responsible for the formation of the intervertebral discs with a long-term goal to develop cell or protein-based therapies for disc degeneration. The Harfe laboratory has extensive expertise in developmental biology and genetics, which allowed us to quickly identify the embryonic notochord as the cell population that forms the entire middle part of the mature disc called the nucleus pulposus (this was one of the aims of the parent grant and this work has now been published). It is this region of the disc that is usually damaged in patients that have severe back pain. In this proposal, we describe pilot experiments to develop a cell-based therapy to heal damaged or degrading discs using purified notochord cells and a model of disc degeneration. This project is a new collaboration between the Harfe laboratory at the University of Florida and the Elliott laboratory at the University of Pennsylvania. The Elliott laboratory has extensive expertise in disc mechanical function and degeneration, animal models of disc degeneration, and disc tissue engineering. Together, the two groups are uniquely equipped to translate our understanding of nucleus pulposus development into cell-based therapies for disc degeneration. The therapeutic opportunity of using cells that normally form the nucleus pulposus to heal damaged discs was identified as a future goal, beyond the 5-year scope, of the parent grant. Due to our recent identification of nucleus pulposus precursor cells it is now possible to begin experiments to directly test possible therapeutic roles for these cells. The interdisciplinary partnership with Dr. Elliott significantly enhances the scope of the parent grant by immediately translating the basic understanding of the developmental mechanism responsible for disc formation discovered by Dr. Harfe to treating disc degeneration. In this proposal we will begin to test the therapeutic potential of notochord cells by first injecting purified notochord cells into the adult nucleus pulposus of normal mice (Aim 1). Our recent exciting discovery that all cells present in the mature mouse nucleus pulposus are derived from the embryonic notochord suggests that notochord cells may be able to function as disc stem cells. To test if purified notochord cells can differentiate into all cell types found in the mature nucleus pulposus we propose to inject these cells, marked with LacZ, into the adult mouse nucleus pulposus. Their ability to survive, proliferates, and form all nucleus pulposus cell types will be determined using molecular markers.

描述（由申请人提供）：据估计，三分之二的美国人一生中至少会经历一次背痛。超过一半的65岁以上的人患有背痛，在美国，每年治疗背痛的费用为500 - 1000亿美元。大多数背痛被认为是由位于每根椎骨之间的椎间盘损伤引起的。目前，对于因物理损伤或退变而受损的椎间盘，还没有治愈方法。母体R01提案的目标是确定负责椎间盘形成的细胞和分子机制，并以开发基于细胞或蛋白质的椎间盘退变治疗为长期目标。Harfe实验室在发育生物学和遗传学方面拥有广泛的专业知识，这使我们能够迅速确定胚胎脊索是形成称为髓核的成熟椎间盘整个中间部分的细胞群（这是父母资助的目标之一，这项工作现已发表）。有严重背痛的患者通常是椎间盘的这个区域受损。在这一建议中，我们描述了利用纯化脊索细胞和椎间盘退变模型开发细胞治疗损伤或退化椎间盘的试点实验。这个项目是佛罗里达大学的Harfe实验室和宾夕法尼亚大学的Elliott实验室的一个新的合作项目。埃利奥特实验室在椎间盘力学功能和退变、椎间盘退变动物模型和椎间盘组织工程方面拥有广泛的专业知识。总之，这两个小组有独特的装备，将我们对髓核发育的理解转化为基于细胞的椎间盘退变治疗。利用通常形成髓核的细胞来治愈受损椎间盘的治疗机会被确定为未来的目标，超出了父母资助的5年范围。由于我们最近鉴定出髓核前体细胞，现在有可能开始实验，直接测试这些细胞可能的治疗作用。与埃利奥特博士的跨学科合作，通过立即将Harfe博士发现的负责椎间盘形成的发育机制的基本理解转化为治疗椎间盘退变，显着提高了父母资助的范围。在这个提议中，我们将开始测试脊索细胞的治疗潜力，首先将纯化的脊索细胞注射到正常小鼠的成年髓核中（目的1）。我们最近令人兴奋的发现，成熟小鼠髓核中存在的所有细胞都来自胚胎脊索，这表明脊索细胞可能具有椎间盘干细胞的功能。为了检验纯化的脊索细胞是否能分化为成熟髓核中发现的所有细胞类型，我们建议将这些标记有LacZ的细胞注射到成年小鼠髓核中。它们的生存、增殖和形成所有髓核细胞类型的能力将通过分子标记来确定。