Notochordal Cell Derived Therapies for Painful Disc Degeneration

描述（由申请人提供）：椎间盘（IVD）退变是一种衰弱性疾病，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。拟议研究的总体目标是引入新的治疗药物和策略，以微创方式限制退变性，恢复IVD结构，减少退变性椎间盘疾病的疼痛状况。目前的治疗方法未能将结构修复和镇痛结合起来。此外，一些镇痛药具有细胞毒性，因此开发新的治疗药物和策略是主要的研究重点。发育动物的大空泡脊索细胞（NCs）协调IVDs，椎骨和周围脊柱结构的模式。人类和其他物种在成年后没有保留nc，表现出与年龄相关的IVD退化，研究人员长期以来一直在寻找nc在人类年轻时丢失的原因。目前的文献表明，nc是一种祖细胞，它们在人体内的早期消失与其向小软骨细胞髓核细胞（SNPCs）分化有关。我们首次拥有了能够将NCs分化为SNPCs的生物反应器和培养方法，因此我们可以从NCs中获得治疗方法并探索其分化机制。这些研究为设计一种基于nc分泌的营养因子的综合治疗干预提供了一种新的范式，以创造一种结构和症状改变疗法，能够恢复IVD功能，并通过抑制神经血管生长到IVD来预防椎间盘源性疼痛。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，这些蛋白质具有因变量，重点是通过限制神经血管侵入和促进结构恢复来抑制疼痛。目的3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评估设计的“鸡尾酒”治疗方法促进合成代谢和抑制椎间盘源性疼痛或疼痛预测因子的效果。这个项目很重要，因为它对椎间盘源性背痛的临床意义重大的问题有潜在的转化作用。该方法具有创新性，因为它研究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这些因素的治疗效果。我们专注于利用新的人体器官培养和大鼠椎间盘源性疼痛模型进行机制测试和筛选研究的治疗潜力。创新和意义也很高，因为确定nc的最佳微环境培养条件将有助于加速对这些未被开发的细胞的研究。

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.