Notochordal Cell Derived Therapies for Painful Disc Degeneration

描述（由申请人提供）：椎间盘（IVD）变性是一种与腰痛发病机制相关的衰弱性疾病，相关医疗费用每年可能超过1000亿美元。拟议研究的总体目标是引入新型治疗药物和策略，以微创方式限制退变，恢复IVD结构，并减少退行性椎间盘疾病的疼痛状况。目前的治疗方法未能整合结构修复和镇痛。此外，几种镇痛剂是细胞毒性的，因此开发新的治疗剂和策略是主要的研究重点。发育中动物的大空泡脊索细胞（NC）协调IVD、椎骨和周围脊柱结构的图案化。人类和其他不保留NC到成年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NC在年轻时在人类中丢失。现在的文献表明，NC是祖细胞，并表明它们在人类中的早期消失与它们分化为小的软骨细胞髓核细胞（SNPC）有关。我们第一次拥有了能够将NCs分化为SNPC的生物反应器和培养方法，以便我们可以从NCs中获得治疗方法并探索其分化机制。拟定的研究提供了一种新的范式，用于设计一种源自NC分泌的营养剂的综合治疗干预，以创建能够恢复IVD功能并通过抑制神经血管生长到IVD中来预防椎间盘源性疼痛的结构和症状改善疗法。目的1将确定能够保持NC表型的微环境条件，表征NC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制性研究，评估NC分泌的蛋白质的治疗潜力，其因变量侧重于通过限制神经血管侵袭和促进结构恢复来抑制疼痛。目的3评价设计的“鸡尾酒”治疗方法在人离体器官培养模型和大鼠体内椎间盘源性疼痛模型中促进和抑制椎间盘源性疼痛或疼痛预测因子的作用。这个项目是重要的，因为翻译潜力的高度临床意义的问题，椎间盘源性背痛。该方法是创新的，因为它调查了发育生物学中的重要因素，并使用描述性和机制研究将其引入治疗效果。我们专注于治疗潜力的机制测试和筛选研究，使用新的人体器官培养和大鼠椎间盘源性疼痛模型。创新性和重要性也很高，因为确定NC的最佳微环境培养条件将有助于加速对这些探索不足的细胞的研究。

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.