Intervertebral Disc Degeneration and Cross-Talk with the Nervous System

椎间盘退变和与神经系统的交互作用

• 批准号: 10897489
• 负责人: Lori A. Setton
• 金额: $ 6.13万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10897489
• 关键词: Action Potentials; Acute; Affect; Afferent Neurons; Algorithms; Anatomy; Animal Model; Animals; Behavior; Behavior monitoring; Behavioral; Brain imaging; Calcium; Cells; Central Nervous System; Chronic; Confocal Microscopy; Dissociation; Distant; Exposure to; Fluorescence Microscopy; Generations; Genetic Transcription; Goals; Harvest; Image; Impairment; In Vitro; Incubated; Infiltration; Inflammation Mediators; Injury; Intervention; Intervertebral disc structure; Ion Channel; Knowledge; Low Back Pain; Measures; Methods; Modeling; Molecular; Molecular Target; Monitor; Morphology; Motion; Mus; Nerve; Nerve Fibers; Nervous System; Neurons; Operative Surgical Procedures; Organ; Organ Culture Techniques; Pain; Pathology; Pattern; Phenotype; Population; Pre-Clinical Model; Puncture procedure; Rattus; Role; Site; Sodium Channel Blockers; Spinal Cord; Spinal Ganglia; TRPV1 gene; Testing; Tetrodotoxin; Tissues; Transgenes; Visualization; Work; afferent nerve; arm; cohort; contrast enhanced; disability; discogenic pain; electric field; fluorophore; in vivo; in vivo fluorescence; injured; intervertebral disk degeneration; microCT; mouse model; neurotransmission; neurotrophic factor; novel; novel strategies; promoter; protein expression; response; second harmonic; sensory stimulus; spinal disk injury; transmission process; two-photon; voltage

Intervertebral disc (IVD) degeneration is one of the greatest contributors to low back pain, yet how the IVD can generate pain remains poorly understood. To date, our knowledge of “cross-talk” between degenerating IVD and sensory nerves involved in transmitting pain is limited to findings of altered protein and RNA expression in tissues of the IVD, the spinal cord or dorsal root ganglia (DRG). Recent advances in the imaging of sensory neuron activation via recording of Ca2+ sensitive fluorescent indicators, together with pre-clinical models of IVD degeneration, now enable the study of temporal and spatial changes in neuronal function and their “cross-talk” to changes in the degenerating IVD. We propose to evaluate action potential-driven Ca2+ transients and molecular changes in sensory neurons in a mouse model of injury-induced IVD degeneration. In Specific Aim 1, we will document temporal changes to pain-related behaviors and sensitivity in Thy1-GCaMP6s mice following puncture of a lumbar IVD to induce IVD degeneration. These mice carry a transgene for the calcium-sensitive fluorophore, GCaMP6s, that is expressed in sensory nerves of the DRG. We will also evaluate the presence of neuronal markers and key ion channels in innervating DRGs, and anatomic changes and nerve fiber infiltration in IVDs, to test for changes with IVD degeneration as compared to sham controls. This work will document molecular changes to IVD and DRGs for this model from 6 to 52 weeks of IVD degeneration, and test for relationships between injured IVD and the innervating lumbar DRGs as a first measure of “cross-talk.” In Specific Aim 2, we will evaluate action potential-driven Ca2+ transients in lumbar DRG neurons in the IVD degeneration model of Specific Aim 1. We will record Ca2+ transients in intact DRG of Thy1-GCaMP6s mice in vitro following electric field stimulation, and measure threshold voltage, 50% maximum (IC50), and numbers of responding DRG neurons and their nearest neighbor response. DRGs will also be tested before and after incubation with sodium channel blockers to screen for remodeling of specific ion channel function with periods of IVD degeneration. Our goal is to identify temporal and spatial changes in DRG function and “cross-talk” with changes of IVD degeneration. Finally, in Specific Aim 3, we will evaluate sensory stimuli-induced responses in the DRG of living mice using in vivo fluorescence microscopy. Working with 2-photon confocal microscopy and motion correction algorithms developed for brain imaging, we will identify the threshold response of DRG neurons in Thy1-GCaMP6s mice with and without IVD degeneration, following in vivo stimulation of brush, pinch, heat and cold. Our goal is to test for relationships between in vivo activation of lumbar DRG neurons with behavioral and sensitivity changes following onset of IVD degeneration. Completion of this study would identify functional changes to sensory neurons at sites distant to the degenerated IVD and reveal new information about IVD- nervous system “cross-talk” that may suggest novel interventions for treatment of discogenic pain.

椎间盘（IVD）变性是导致腰痛的最大因素之一，但是IVD如何 产生的疼痛仍然很少理解。迄今为止，我们对IVD变性和 涉及传播疼痛的感觉神经仅限于组织中蛋白质改变和RNA表达的发现 IVD，脊髓或背根神经节（DRG）。感觉神经元成像的最新进展 通过记录Ca2+敏感荧光指标的激活，以及IVD的临床前模型 退化，现在可以研究神经元功能的暂时和空间变化及其“交叉对话” 变性IVD。 我们建议评估动作电位驱动的Ca2+瞬变和感觉的分子变化 损伤引起的IVD变性的小鼠模型中的神经元。在特定目标1中，我们将记录 对疼痛相关行为的暂时变化和thy1-Gcamp6s小鼠的敏感性刺穿后 腰部IVD诱导IVD变性。这些小鼠对钙敏感的荧光团具有转换， GCAMP6在DRG的感觉神经中表达。我们还将评估神经元的存在 神经DRG的标记和关键离子通道，以及IVD中的解剖学变化和神经纤维浸润， 与假对照相比，测试IVD变性的变化。这项工作将记录分子 该模型的IVD和DRG的更改从6周到52周的IVD变性，并测试关系 在受伤的IVD和神经支配的腰椎DRG之间，作为“串扰”的第一个量度。在特定的目标2中，我们 将评估在IVD变性模型中，腰椎DRG神经元中的动作电位驱动的Ca2+瞬变 特定目标1。我们将在电气后在体外记录Thy1-GCAMP6S小鼠的完整DRG中的Ca2+瞬变 现场仿真和测量阈值电压，最大50％（IC50）和响应DRG数量 神经元及其最近的邻居反应。在与钠孵育之前和之后还将测试DRG 通道阻滞剂以筛选使用IVD变性周期的特定离子通道函数进行重塑。我们的 目标是确定DRG功能的暂时和空间变化以及与IVD变化的“串扰” 退化。最后，在特定的目标3中，我们将评估感官刺激诱导的响应 使用体内荧光显微镜的活小鼠。使用2光子共聚焦显微镜和运动 为大脑成像开发的校正算法，我们将确定DRG神经元在 在体内刺激刷子，捏，热和 寒冷的。我们的目标是测试与行为和行为和 IVD变性发作后灵敏度变化。这项研究的完成将确定功能 变化的位点的感觉神经元变化，并揭示了有关IVD-的新信息 神经系统“串扰”可能暗示了治疗椎间盘痛的新干预措施。

项目成果

Contrast-enhanced microCT evaluation of degeneration following partial and full width injuries to the mouse lumbar intervertebral disc.

• DOI: 10.1038/s41598-022-19487-9
• 发表时间: 2022-09-16
• 期刊: SCIENTIFIC REPORTS
• 影响因子: 4.6
• 作者: Walk, Remy E.;Moon, Hong Joo;Tang, Simon Y.;Gupta, Munish C.
• 通讯作者: Gupta, Munish C.

Integrin-mediated interactions with a laminin-presenting substrate modulate biosynthesis and phenotypic expression for cells of the human nucleus pulposus.

• DOI: 10.22203/ecm.v041a50
• 发表时间: 2021-06-24
• 期刊: European cells & materials
• 影响因子: 3.1
• 作者: Speer J;Barcellona M;Jing L;Liu B;Lu M;Kelly M;Buchowski J;Zebala L;Luhmann S;Gupta M;Setton L
• 通讯作者: Setton L

Analysis of Infiltrating Immune Cells Following Intervertebral Disc Injury Reveals Recruitment of Gamma-Delta (γδ) T cells in Female Mice.

椎间盘损伤后浸润免疫细胞的分析揭示了雌性小鼠中 Gamma-Delta (γδ) T 细胞的募集。

• DOI: 10.1101/2024.03.01.582950
• 发表时间: 2024
• 期刊: bioRxiv : the preprint server for biology
• 作者: Clayton,SadeW;Walk,RemyE;Mpofu,Laura;Easson,GarrettWD;Tang,SimonY
• 通讯作者: Tang,SimonY

Bioactive in situ crosslinkable polymer-peptide hydrogel for cell delivery to the intervertebral disc in a rat model.

• DOI: 10.1016/j.actbio.2021.06.045
• 发表时间: 2021-09-01
• 期刊: ACTA BIOMATERIALIA
• 影响因子: 9.7
• 作者: Barcellona, Marcos N.;Speer, Julie E.;Jing, Liufang;Patil, Deepanjali S.;Gupta, Munish C.;Buchowski, Jacob M.;Setton, Lori A.
• 通讯作者: Setton, Lori A.

Assessment of bovine cortical bone fracture behavior using impact microindentation as a surrogate of fracture toughness.

使用冲击微压痕作为断裂韧性的替代物来评估牛皮质骨骨折行为。

• DOI: 10.1101/2023.08.07.552351
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Jahani,Babak;Vaidya,Rachana;Jin,JamesM;Aboytes,DonaldA;Broz,KaitlynS;Khrotapalli,Siva;Pujari,Bhanuteja;Baig,WaleeM;Tang,SimonY
• 通讯作者: Tang,SimonY