Mechanisms for Regenerative Healing in Intervertebral Discs

椎间盘再生愈合机制

• 批准号: 10344363
• 负责人: James C. Iatridis
• 金额: $ 56.65万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-15 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10344363
• 关键词: Adult; Affect; Age; Aging; Anatomy; Atomic Force Microscopy; Back Pain; Biomechanics; Caliber; Cell Maturation; Cell Proliferation; Cell Therapy; Cells; Chronic; Clinical Trials; Collagen; Data; Defect; Deposition; Engineering; Environment; Extracellular Matrix; Fibrosis; Functional disorder; Genetic; Gold; Growth; Height; Human; Inflammation; Injections; Injury; Integrins; Intervertebral disc structure; Ligands; Mediating; Microscopy; Mitosis; Mitotic; Modeling; Modulus; Mus; Natural regeneration; Neonatal; Nerve; Outcome; Pain; Phenotype; Play; Population; Population Heterogeneity; Process; Property; Protein-Lysine 6-Oxidase; Puncture procedure; Recurrence; Regenerative Medicine; Regenerative capacity; Role; Thinking; Time; Tissue Engineering; Tissues; beta Aminopropionitrile; cell regeneration; crosslink; density; design; disability; disability impact; discogenic pain; effective therapy; ethylene glycol; healing; improved; inhibitor; innovation; intervertebral disk degeneration; mouse model; nanoindentation; neonate; neurovascular; novel; nucleus pulposus; postnatal; progenitor; recruit; regeneration potential; regenerative; regenerative repair; repair strategy; restoration; scleraxis; skeletal; standard care; stem cells; tool; transcriptome sequencing; treatment strategy

Summary Intervertebral disc (IVD) degeneration contributes to ~40% of back pain cases. Structural IVD defects distinguish degeneration from aging and play a role pain and disability. There is a critical unmet need for improved annulus fibrosus (AF) repair strategies since discectomy, the gold standard treatment for removing herniated nucleus pulposus (NP) tissue from AF defects, leaves AF defects unrepaired and complications include reherniation and recurrent degeneration-related pain. While clinical trials of IVD cell therapy show promise to reduce discogenic pain and disability they do not involve optimized delivery strategies, and are not informed by natural IVD healing processes since remarkably little is known about the diversity of AF cell populations and their roles in healing. We believe an IVD regenerative healing model is required to identify successful AF healing strategies and to identify cellular and micromechanical factors critical in successful healing to serve as a roadmap for regenerative medicine treatments. We've developed a successful regenerative AF healing model in mice and show neonatal IVDs with severe AF puncture heal with complete restoration of IVD height and biomechanical properties while adults heal fibrotic deposition and loss of IVD height and biomechanical function. The premise of this project is that neonates regeneratively heal while skeletally mature mice do not due to increased extracellular matrix (ECM) stiffness and altered ligand presentation resulting in terminal differentiation of AF progenitors. Aim 1 determines effects of growth, maturation, and matrix stiffness on IVD healing and determines when the regenerative window closes. We apply mouse models to determines the postnatal age that the AF regenerative healing window closes, if complete AF structural regeneration is possible, and if altering ECM stiffness can extend the regenerative healing window and prolong the age when AF cells are in mitosis. Aim 2 identifies distinct AF progenitor populations, their loss with maturation, and roles of these progenitors in healing. We use single cell and spatial sequencing in mouse IVDs to identify distinct AF cell populations and their localization in mice of regenerative healing, post-regenerative healing, and regenerative restoration groups. Aim 3 engineers a soft-synthetic substrate that promotes immature AF cell phenotypes. We identify design criteria in mouse and human ECM and cells and control substrate stiffness, ligand type, and density using functionalized poly(ethylene glycol) substrates. Outcomes of this project include determining when the IVD regenerative repair window closes and if full regeneration is possible; identifying disperse AF progenitor populations and their roles in regenerative healing; and determining critical design factors that promote immature AF progenitor phenotypes and inform cell delivery strategies.

概括 椎间盘（IVD）变性约为背痛病例的40％。结构IVD缺陷 区分退化与衰老并发挥作用疼痛和残疾。有关键的未满足需求 自盘切除术以来，改进的环纤维（AF）维修策略是去除的金标准处理 椎间盘突出症核（NP）组织因AF缺陷，AF缺陷未经修复和并发症 包括重新连接和复发性变性有关的疼痛。 IVD细胞疗法的临床试验显示 承诺减轻椎间盘源性疼痛和残疾，它们不涉及优化的交付策略，而不是 由天然IVD愈合过程所告知，因为对AF细胞的多样性知之甚少 人群及其在康复中的作用。我们认为需要一个IVD再生愈合模型来识别 成功的AF治疗策略并确定成功的细胞和微机械因素 作为再生医学治疗方法的路线图。我们已经开发了成功 小鼠的再生AF愈合模型，并显示出严重的AF穿刺治疗的新生儿IVD 恢复IVD高度和生物力学特性，而成年人治愈纤维化沉积和IVD的损失 高度和生物力学功能。该项目的前提是新生儿再生治愈 骨骼成熟的小鼠并不是由于细胞外基质（ECM）刚度增加和配体改变 演示导致AF祖细胞的终末分化。 AIM 1决定增长的影响， IVD愈合上的成熟和基质刚度，并确定再生窗口何时关闭。我们 应用鼠标模型来确定AF再生愈合窗口关闭的产后年龄，如果 完全AF结构再生是可能的，如果改变ECM刚度可以扩展再生 治愈窗口并延长AF细胞处于有丝分裂的年龄。 AIM 2标识不同的AF祖先 人口，成熟的损失以及这些祖细胞在康复中的作用。我们使用单元和空间 在小鼠IVD中进行测序以鉴定不同的AF细胞群体及其在再生小鼠中的定位 愈合，后再生愈合和再生恢复组。 AIM 3工程师软合伙 促进未成熟AF细胞表型的底物。我们确定鼠标和人类ECM中的设计标准 细胞和控制底物刚度，配体类型和密度使用官能化的聚乙二醇） 基材。该项目的结果包括确定IVD再生维修窗口何时关闭以及 如果可以完全再生；确定分散的AF祖细胞及其在再生中的作用 康复;并确定促进未成熟AF祖细胞表型并告知的关键设计因素 细胞输送策略。