Stratification of nucleus pulposus cell sub-populations during intervertebral disc development, ageing and degeneration

椎间盘发育、老化和退变过程中髓核细胞亚群的分层

• 批准号: 2283720
• 金额: --
• 依托单位: University of Manchester
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2283720
• 关键词: Stratification; nucleus; pulposus; cell; sub

Back pain is a leading cause of disability in the UK and intervertebral disc (IVD) degeneration has been identified as the cause in a high proportion of patients. As current therapies for chronic back pain demonstrate poor long-term efficacy, there is an urgent clinical need for advanced cell-based regenerative therapies. However, an essential step in development of such therapies is a detailed understanding of native cell biology and how this changes with development, ageing and degeneration. The central nucleus pulposus (NP) region of the IVD is formed from the embryonic notochord as it segments during foetal development, while the surrounding ligamentous annulus fibrosus (AF) region is formed from the sclerotome/mesoderm. At birth, the NP is populated by morphologically distinct, large vacuolated notochordal cells (NCs), but these cells gradually disappear during skeletal maturation eventually becoming undetectable and replaced by a population of smaller round cells - i.e. the NP cells. This change in cell population correlates with initiation of degenerate changes within the matrix suggesting that loss of NC cells may be responsible for degeneration and ultimately back pain. However, it is currently unclear whether there is persistence of NCs or other progenitor cells in the adult human IVD, or how the cellular composition of the NP changes with development, ageing and degeneration. Elucidation of such changes will allow identification of anabolic cell sub-populations which may be critical to IVD homeostasis and as such may have potential in regenerative therapies.The lab has recently undertaken phenotypic profiling of both human foetal NCs and adult NP cells, which have identified potential markers of NC and NP populations and have preliminary evidence that subpopulations of cells exist with the adult IVD. They have also identified changes in phenotype of adult NP cells during degeneration, including evidence of cellular senescence and adoption of a catabolic phenotype. The overall aims of this study are: (i) to use a panel of novel phenotypic markers to identify and isolate cell sub-populations from the human IVD during different stages of foetal development, adolescent skeletal maturation, and ageing/degeneration; and (ii) to compare the phenotype and function of isolated sub-populations to establish whether a population exists which has regenerative potential. These aims will be achieved through a range of state-of-the-art techniques, including CyTOF mass cytometry, RNAseq transcriptomic profiling and functional in vitro human cell studies to elucidate the phenotype and behaviour of human NP cells across development, skeletal maturation, ageing and degeneration.

在英国，背痛是导致残疾的主要原因，椎间盘 (IVD) 退变已被确定为大部分患者的病因。由于目前治疗慢性背痛的长期疗效较差，因此临床迫切需要先进的基于细胞的再生疗法。然而，开发此类疗法的一个重要步骤是详细了解天然细胞生物学以及其如何随着发育、衰老和退化而变化。 IVD 的中央髓核 (NP) 区域由胚胎脊索在胎儿发育过程中分段形成，而周围的韧带纤维环 (AF) 区域由巩膜/中胚层形成。出生时，NP 中充满了形态独特的大型空泡脊索细胞 (NC)，但这些细胞在骨骼成熟过程中逐渐消失，最终变得无法检测到，并被一群较小的圆形细胞（即 NP 细胞）所取代。细胞群的这种变化与基质内退化变化的开始相关，表明 NC 细胞的损失可能是退化并最终导致背痛的原因。然而，目前尚不清楚成人 IVD 中是否存在 NC 或其他祖细胞的持续存在，或者 NP 的细胞组成如何随着发育、衰老和退化而变化。阐明这些变化将有助于识别合成代谢细胞亚群，这些细胞亚群可能对 IVD 稳态至关重要，因此可能在再生疗法中具有潜力。该实验室最近对人类胎儿 NC 和成人 NP 细胞进行了表型分析，确定了 NC 和 NP 群体的潜在标记，并初步证据表明成人 IVD 中存在细胞亚群。他们还发现了成年 NP 细胞在变性过程中表型的变化，包括细胞衰老和分解代谢表型的证据。本研究的总体目标是：（i）使用一组新型表型标记物来识别和分离胎儿发育、青少年骨骼成熟和衰老/退化不同阶段的人类 IVD 细胞亚群； (ii) 比较分离的亚群体的表型和功能，以确定是否存在具有再生潜力的群体。这些目标将通过一系列最先进的技术来实现，包括 CyTOF 质谱流式分析、RNAseq 转录组分析和功能性体外人类细胞研究，以阐明人类 NP 细胞在发育、骨骼成熟、衰老和退化过程中的表型和行为。