Notochordal Cell Derived Therapies for Painful Disc Degeneration

描述(申请人提供)：腰椎间盘(IVD)退行性变是一种衰弱的疾病，与腰痛的发病机制有关，相关的医疗费用每年可超过1000亿美元。这项拟议研究的总体目标是引入新的治疗药物和策略，以微创的方式使用，以限制退变，恢复IVD结构，并减少退行性间盘疾病的痛苦状况。目前的治疗方法未能将结构修复和止痛结合起来。此外，几种镇痛剂具有细胞毒性，因此开发新的治疗药物和策略是主要的研究重点。发育中的动物体内的大液泡化脊索细胞(NCs)协调着IVD、椎骨和周围脊柱结构的图案。人类和其他成年后不保留NC的物种表现出与年龄相关的IVD退化，研究人员长期以来一直试图回答为什么NCS在人类年轻时就消失了。现在的文献表明，NCs是祖细胞，并提示它们在人类中的早期消失与它们向小软骨细胞髓核细胞(SNPC)的分化有关。我们第一次拥有了生物反应器和培养方法，能够将NCs分化为SNPC，这样我们就可以从NCs中获得治疗方法，并探索它们分化的机制。这些研究为设计一种从NCS分泌的营养物质衍生的综合治疗干预提供了新的范式，以创造能够恢复IVD功能并通过抑制进入IVD的神经血管生长来预防间盘源性疼痛的结构和症状修改疗法。目的1确定保持NC表型的微环境条件，鉴定NCS产生的生物活性分子的重要基因和蛋白质的表型稳定性，并分离参与NC分化的途径。目的2是一系列描述性和机械性研究，用因变量评估NCS分泌的蛋白质的治疗潜力，这些变量集中在通过限制神经血管的侵袭和促进结构恢复来抑制疼痛。目的在人体外器官培养模型和大鼠在体盘源性疼痛模型中，评价设计的“鸡尾酒”疗法促进合成代谢和抑制盘源性疼痛或疼痛预测因子的作用。这个项目意义重大，因为它有可能转化为具有高度临床意义的间盘性背痛问题。这种方法是创新的，因为它调查了发育生物学中的重要因素，并通过描述性和机械性研究介绍了它们的治疗效果。我们使用新的人体器官培养和大鼠关节盘源性疼痛模型，通过机械测试和筛选研究，专注于治疗潜力。创新和意义也很高，因为确定NCs的最佳微环境培养条件将有助于加速对这些未被探索的细胞的研究。

DESCRIPTION (provided by applicant): Intervertebral disc (IVD) degeneration is a debilitating disorder implicated in the pathogenesis of low back pain with associated medical costs that can exceed $100 billion annually. The overall goal of the proposed research is to introduce novel therapeutic agents and strategies for use in a minimally invasive manner to limit degeneration, restore IVD structure, and reduce painful conditions of degenerative disc disease. Current therapies fail to integrate both structural repair and analgesia. Further, several analgesics are cytotoxic so that developing new therapeutic agents and strategies are a major research priority. The large vacuolated notochordal cells (NCs) in developing animals orchestrate patterning of the IVDs, vertebrae and surrounding spinal structures. Humans and other species that do not retain NCs into adulthood exhibit age related IVD degeneration and researchers have long sought to answer why NCs are lost in humans at young ages. The literature now indicates NCs are progenitor cells and suggests their early disappearance in humans is associated with their differentiation to small chondrocytic nucleus pulposus cells (SNPCs). For the first time, we have a bioreactor and culture methods capable of differentiating NCs into SNPCs so that we can derive therapies from NCs and explore mechanisms for their differentiation. The proposed studies provide a new paradigm for designing an integrated therapeutic intervention derived from trophic agents secreted by NCs to create structure and symptom modifying therapies capable of restoring IVD function and preventing discogenic pain by inhibition of neurovascular growth into the IVD. Aim 1 will determine microenvironment conditions capable of retaining NC phenotype, characterize the phenotypic stability of important genes and proteins of bioactive molecules produced by NCs, and isolate pathways involved in NC differentiation. Aim 2 is a series of descriptive and mechanistic studies that assess therapeutic potential of proteins secreted by NCs with dependent variables that focus on pain inhibition by limiting neurovascular invasion and promoting structural restoration. Aim 3 evaluates designed 'cocktail' treatments for their effects promoting anabolism and inhibiting discogenic pain or predictors of pain in human ex vivo organ culture models and rat in vivo discogenic pain models. This project is significant because of the translational potential to the highly clinically significant problem of discogenic back pain. The approach is innovative because it investigates factors important in developmental biology and introduces them for therapeutic effect using descriptive and mechanistic studies. We focus on therapeutic potential with mechanistic testing and screening studies using novel human organ culture and rat discogenic pain models. Innovation and significance are also high because determining optimal microenvironmental culturing conditions of NCs will help accelerate the growing body of research on these underexplored cells.