Targeting cell senescence in a novel model of spontaneous disc degeneration

在自发性椎间盘退变的新模型中靶向细胞衰老

基本信息

批准号：
10839574
负责人：
Makarand V Risbud
金额：
$ 7.97万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2023-11-10
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10839574
关键词：
Acute Address Affect American Animal Model Back Biological Candidate Disease Gene Canis familiaris Cell Aging Cell Survival Cell physiology Cells ChIP-seq Chronic Dasatinib Data Disease Etiology Excision Exhibits Gene Expression Genes Genetic Genetic Variation Genotype Gerbils Goals Health Health Status Human Inbred Strains Mice Inflammatory Injury Intervention Intervertebral disc structure LG/J Mouse Link Low Back Pain Measures Metabolic Metabolism Modeling Modification Molecular Mouse Strains Mus Mutation Neck Neck Pain Opioid Organ Culture Techniques Pathogenesis Pathology Pathway interactions Persons Pharmaceutical Preparations Phenotype Pilot Projects Population Prevalence Process Publishing Quantitative Trait Loci Quercetin Reporting Resolution Risk Factors SM/J Mouse SNP array Sand Rats Structure Testing Time Tissue Preservation Tissues Traumatic injury Variant Vertebral column burden of illness early onset experience genetic manipulation genome wide association study in vivo insight insurance claims intervertebral disk degeneration metabolic profile metabolomics mouse genome mouse model novel nucleus pulposus opioid abuse opioid mortality prevent regenerative regenerative tissue senescence therapeutic development trait transcriptome transcriptome sequencing transcriptomics years lived with disability

项目摘要

Intervertebral disc degeneration is the major risk factor associated with chronic neck and low back pain, ubiquitous health conditions that affects millions of people world-wide. There is an incomplete understanding of the pathogenesis of disc degeneration partially due to lack of an appropriate animal model. Current animal models primarily use traumatic insults to promote degeneration differing from most human cases of disc degeneration. While, a few animal models of spontaneous disc degeneration are reported, several limitations prevent their wide-spread use. Thus, there is a great need for small animal models that are more representative of human disc pathology. Previous studies have investigated inbred strains of mice including SM/J and LG/J for their regenerative ability. In contrast to LG/J, a “super healer” strain, SM/J was found to be a “poor healer”. However, there no studies have investigated the disc health status in these strains of mice. We show for the first time that SM/J mice exhibit spontaneous disc degeneration that captures important features of human disc pathology. Based on our novel pilot data, we hypothesize that this degenerative phenotype has a strong genetic basis and driven by early changes in cell phenotype, metabolism and senescence. We will investigate the cellular and genetic mechanisms that underlie disc degeneration in SM/J mice in three Aims. In Aim 1 we will test the hypothesis that disc degeneration in SM/J mice is characterized by compromised cell survival, altered cell phenotype, metabolism and senescence. We will perform RNA-Seq and ChIP-Seq analysis of NP tissue of SM/J and LG/J mice. We will also perform metabolomics to determine metabolic status of cells. These studies will provide unbiased insights into temporal alterations in transcriptome and metabolism to delineate pathways and mechanisms central to the pathogenesis of disc degeneration in SM/J mice. In Aim 2 we will test the hypothesis that removal of senescent cells from the intervertebral disc slows down the progression of disc degeneration in SM/J mice. Pilot studies have shown that there is an accumulation of senescent cells in SM/J discs. We will treat ex vivo organ cultured discs and SM/J mice in vivo during early and established stages of disc degeneration with well-characterized senolytic drugs and analyze metabolic, histopathological and genetic changes. Lastly in Aim 3 we will delineate the genetic mechanisms that contribute to cell senescence and disc degeneration in SM/J mice. We will perform a genome-wide association study (GWAS) using LG/J x SM/J Advanced Intercross Lines (AILs) to discover regions of the mouse genome contributing to differences in disc degeneration. For this purpose, AIL mice will be genotyped at 143,000 GIGAmuga chip single nucleotide polymorphisms (SNPs) and discs from several mice will be used for RNA-Seq analysis. Together, these data will be used to identify high-resolution quantitative trait loci (QTL) for disc traits ~5 genes per support interval. The genes responsible for these QTL will be identified. This aim will generate candidate genes likely contributing to variation in the senescence and disc degeneration between LG/J and SM/J strains.

椎间盘退变是慢性颈痛和腰痛的主要危险因素，影响全世界数百万人的普遍健康状况目前尚不完全了解。椎间盘退变发病机制的部分原因是缺乏合适的动物模型。与大多数人类椎间盘病例不同，模型主要使用创伤性损伤来促进退化虽然有一些自发性椎间盘退变的动物模型被报道，但也存在一些局限性。因此，非常需要更具代表性的小动物模型。先前的研究已经调查了小鼠的近交系，包括 SM/J 和 LG/J。与“超级治疗者”菌株 LG/J 相比，SM/J 被认为是“较差的治疗者”。然而，我们首次展示了没有研究调查这些小鼠品系的椎间盘健康状况。 SM/J 小鼠表现出自发性椎间盘退变的时间，捕捉了人类椎间盘的重要特征根据我们的新试验数据，我们发现这种退行性表型具有很强的遗传性。基础并由细胞表型、代谢和衰老的早期变化驱动。在目标 1 中，我们将在三个目标中研究 SM/J 小鼠椎间盘退变的细胞和遗传机制。检验 SM/J 小鼠椎间盘退变的特征是细胞存活受损的假设，我们将对 NP 组织进行 RNA-Seq 和 ChIP-Seq 分析。我们还将进行 SM/J 和 LG/J 小鼠的代谢组学来确定细胞的代谢状态。将为转录组和代谢的时间变化提供公正的见解，以描绘在目标 2 中，我们将探讨 SM/J 小鼠椎间盘退变发病机制的核心途径和机制。检验以下假设：从椎间盘中去除衰老细胞会减慢衰老的进展初步研究表明，SM/J 小鼠的椎间盘退化存在衰老细胞的积累。我们将在早期和建立阶段治疗离体器官培养的椎间盘和体内 SM/J 小鼠。使用特征良好的 senolytic 药物分析椎间盘退变的阶段，并分析代谢、组织病理学最后，在目标 3 中，我们将描述影响细胞的遗传机制。 SM/J 小鼠的衰老和椎间盘退变我们将进行全基因组关联研究 (GWAS)。使用 LG/J x SM/J 高级交叉系 (AIL) 发现小鼠基因组中有助于为此，将在 143,000 GIGAmuga 芯片上对 AIL 小鼠进行基因分型。来自几只小鼠的单核苷酸多态性 (SNP) 和圆盘将用于 RNA 测序分析。这些数据将共同用于识别圆盘性状约 5 个基因的高分辨率数量性状基因座 (QTL) 每个支持间隔将确定负责这些 QTL 的基因。可能导致 LG/J 和 SM/J 菌株之间衰老和椎间盘退变变异的基因。