The role of notochord derived signaling, mechanical force generation and AF derived Tgfβ signaling on intervertebral disc formation

脊索衍生信号、机械力产生和 AF 衍生 Tgfβ 信号对椎间盘形成的作用

• 批准号: 10414098
• 负责人: Rose Long
• 金额: $ 7.17万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10414098
• 关键词: Actins; Back; Biological Assay; Cell Proliferation; Cells; Cessation of life; Chondrocytes; Collagen; Collagen Fiber; Complex; Cues; Deposition; Development; Diphtheria Toxin; Enterobacteria phage P1 Cre recombinase; Extracellular Matrix; Fiber; Future; Generations; Goals; Grant; Growth Factor; Immunohistochemistry; In Situ Hybridization; In Vitro; Intervertebral disc structure; Low Back Pain; LoxP-flanked allele; Measures; Mechanical Stress; Mechanics; Microfilaments; Molecular; Movement; Mus; Myosin ATPase; Myosin Type II; Outcome; Pain; Pathway interactions; Pharmacology; Process; Production; Proteins; Proteoglycan; Role; SHH gene; Sclerotome; Signal Pathway; Signal Transduction; Somites; Sonication; Source; Stimulus; Stress; Stress Fibers; Structure; Testing; Time; Tissue Engineering; Tissues; Transcript; Transforming Growth Factor beta; Work; cartilaginous; cytotoxic; directed differentiation; disability; extracellular; fiber cell; functional restoration; improved; insight; intervertebral disk degeneration; mechanical force; mechanical load; molecular phenotype; morphogens; mouse genetics; notochord; nucleus pulposus; postnatal; receptor; regenerative; regenerative approach; regenerative biology; regenerative repair; repair strategy; repaired; rho; small molecule; spinal disk injury; stress state; therapeutic target; transcriptome sequencing; vertebra body

PROJECT SUMMARY / ABSTRACT Intervertebral disc (IVD) injury such as herniation and degeneration can result in low back pain which is a leading cause of global disability. Tissue engineering strategies which successfully recapitulate IVD structure could restore function and reduce the burden of IVD injury. However, intervertebral disc development is not well understood, hindering the therapeutic targeting of key molecular factors. Initially, the IVD center, the nucleus pulposus, arises from the segmentation of the continuous notochord through the movement of notochordal cells out of the future vertebra body and into the intervertebral disc. The IVD fibrous exterior, the annulus fibrosus, arises from sclerotomal cells which surround the notochord and differentiatiate through the deposition of aligned collagen fibers production of extracellular matrix. Molecularly, the notochord is a source of the secreted morphogen Sonic Hedghehog (Shh), and annulus fibrosus expresses the growth factor Tgfβ. This grant will test the necessity of the notochord in two different stages of IVD development: proliferation, and actin fiber formation in the AF. The second aim will determine the role of the mechanical force generation, myosin, actin filament formation and Rho pathway activation on AF formation. The last aim determines the quantitiative expression and necessity of Tgfβ signaling in AF differentiation including cell alignment and extracellular matrix deposition. This project utilizes mouse genetics to manipulate molecular cues, and assays the molecular and phenotypic effects with in situ hybridization, immunohistochemistry and RNA sequencing. The successful completion of the project will provide insights on molecular and mechanical drivers of intervertebral disc formation and yield insights which can be used for developing repairs for painful IVD injury.

项目摘要/摘要 椎间盘(IVD)损伤，如突出和退变，可导致下腰痛，这是一种 全球残疾的主要原因。成功重现IVD结构的组织工程策略 可恢复功能，减轻IVD损伤负担。然而，椎间盘的发育并不是 众所周知，阻碍了关键分子因子的治疗靶向。最初，IVD中心， 髓核是由连续脊索的运动引起的 脊索细胞从未来的椎体进入椎间盘。IVD纤维外观， 纤维环起源于脊索周围的硬膜细胞，并通过 排列整齐的胶原纤维沉积，产生细胞外基质。从分子上讲，脊索是一种来源 分泌形态的Sonic Hedghehog(Shh)和纤维环表达生长因子转化生长因子β。 这笔赠款将在IVD发展的两个不同阶段测试脊索的必要性：增殖，和 房颤时肌动蛋白纤维的形成。第二个目标将决定机械力产生的作用， 肌球蛋白、肌动蛋白细丝形成和Rho通路激活对房颤形成的影响。最后一个目标决定了 转化生长因子β信号在房颤分化中的定量表达及必要性 细胞外基质沉积。这个项目利用老鼠的遗传学来操纵分子线索，并分析 用原位杂交、免疫组织化学和RNA测序研究分子和表型效应。 该项目的成功完成将提供关于分子和机械驱动因素的见解 这是一项重要的研究成果，可用于对疼痛的IVD损伤进行修复。