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.

椎间盘变性是与慢性颈和下背痛有关的主要危险因素，无处不在的健康状况会影响全球数百万人。有一个不完整的理解椎间盘变性的发病机理部分由于缺乏适当的动物模型。目前的动物模型主要使用创伤性损伤以促进退化与大多数人类椎间盘病例的区别退化。虽然报道了一些赞助椎间盘变性的动物模型，但有几个限制防止其广泛使用。这是对更具代表性的小动物模型的巨大需求人类椎间盘病理学。先前的研究研究了包括SM/J和LG/J在内的小鼠的近交菌株因为它们的再生能力。与“超级治疗师”菌株LG/J相反，SM/J被发现是“可怜的治疗师”。但是，没有研究调查这些小鼠菌株中的椎间盘健康状况。我们展示第一个 SM/J小鼠表现出赞成的圆盘变性的时间，捕获了人盘的重要特征病理。根据我们新颖的试点数据，我们假设这种退化表型具有很强的遗传基础和由细胞表型，代谢和感应的早期变化驱动。我们将调查三个目标中SM/J小鼠的椎间盘变性的细胞和遗传机制。在目标1中，我们将检验以下假设，即SM/J小鼠中的椎间盘变性的特征是细胞存活受损，改变了细胞表型，代谢和感应。我们将对NP组织的RNA-SEQ和CHIP-SEQ分析 SM/J和LG/J小鼠。我们还将执行代谢组学以确定细胞的代谢状态。这些研究将提供对转录组和新陈代谢暂时改变的无偏见 SM/J小鼠椎间盘变性的发病机理的途径和机制。在目标2中，我们将检验以下假设：从椎间盘中去除感觉细胞会减慢 SM/J小鼠的椎间盘变性。试点研究表明，在 SM/J光盘。我们将在早期和确定的体内进行体内组织的培养碟和SM/J小鼠椎间盘变性的阶段与特征良好的鼻溶液药物并分析代谢，组织病理学和遗传变化。最后，在AIM 3中，我们将描述有助于细胞的遗传机制 SM/J小鼠的感应和椎间盘变性。我们将进行全基因组协会研究（GWAS）使用LG/J X SM/J高级间交叉线（AILS）来发现小鼠基因组的区域椎间盘变性的差异。为此，将AIL小鼠以143,000 Gigamuga芯片进行基因分型来自几只小鼠的单核苷酸多态性（SNP）和椎间盘将用于RNA-Seq分析。共同使用这些数据来识别椎间盘特征〜5基因的高分辨率定量性状基因座（QTL）每个支持间隔。将确定负责这些QTL的基因。这个目标将产生候选人基因可能导致LG/J和SM/J菌株之间的感应和椎间盘变性变化。