Function of Fibro-Adipogenic Progenitors in Heterotopic Ossification of Skeletal Muscle

纤维脂肪祖细胞在骨骼肌异位骨化中的功能

基本信息

批准号：
10218059
负责人：
DAVID J GOLDHAMER
金额：
$ 33.5万
依托单位：
UNIVERSITY OF CONNECTICUT STORRS
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10218059
关键词：
ACVR1 gene Ablation Activins Address Affect Alleles Amino Acids Apoptosis Area BMP2 gene Blocking Antibodies Bone Formation Inhibition Bone Growth Candidate Disease Gene Cartilage Cell Communication Cell Proliferation Cell Therapy Cells Coculture Techniques Cre driver Development Diphtheria Toxin Disease Dominant Genetic Conditions Enhancers Environment Exhibits Fiber Fibrosis Functional disorder Gene Expression Gene Expression Profile Gene Targeting General Population Genetic Diseases Genetic Models Genetic Transcription Glycine Goals Health Heterotopic Ossification Hypersensitivity Immunofluorescence Immunologic Impairment Individual Infiltration Injections Injury Intramuscular Injections Kinetics Lead Life Life Expectancy Ligands Mesenchymal Stem Cells Messenger RNA Modeling Molecular Morbidity - disease rate Mus Muscle Muscle satellite cell Muscular Atrophy Natural regeneration Osteogenesis Pathogenesis Pathologic Pathway interactions Patients Physiologic Ossification Physiological Polymerase Population Quality of life RNA analysis Regenerative capacity Research Running Serine Signal Transduction Skeletal Muscle Source Specificity Therapeutic Time Traumatic injury Wild Type Mouse activin A alpha Toxin base bone bone morphogenetic protein receptors experimental study genome-wide global run on sequencing muscle regeneration mutant osteogenic prevent progenitor progressive myositis ossificans promoter response satellite cell soft tissue tibialis anterior muscle transcriptome transcriptome sequencing

项目摘要

Project Summary Heterotopic ossification (HO), the formation of bone in skeletal muscle and associated soft tissues, can result from traumatic injury or disease. The most extreme form of HO is manifested in the rare, autosomal- dominant genetic disorder, Fibrodysplasia ossificans progressiva (FOP), in which HO continues progressively throughout life, resulting in devastating effects on health and life expectancy. We developed a new genetic model of FOP based on conditional expression of the disease-causing BMP receptor, Acvr1(R206H). Using this model, we identified fibro-adipogenic progenitors (FAPs), a multipotent mesenchymal progenitor in muscle tissue, as the disease-causing cell population. Notably, we have shown that intramuscular injection of BMP2 into wild type mice also leads to FAP-derived HO, suggesting mechanistic similarities of HO pathogenesis in FOP patients and the general population. Studies of HO have focused almost entirely on mechanisms of bone growth, yet severe muscle loss can be a significant contributing factor to patient morbidity. The overarching objectives of this research are to 1) understand how FAP programming becomes subverted for pathological bone formation; 2) understand how regeneration is inhibited in skeletal muscle susceptible to HO; 3) determine whether HO of skeletal muscle is entirely dependent on FAPs. By targeting Acvr1R206H expression to FAPs using PdgrfαCreER and Tie2-Cre drivers, Aim 1 will quantify regeneration impairment, and will determine whether muscle stem cell (satellite cell; SC) dysfunction is responsible for regeneration deficits. These studies will quantify SC proliferation and apoptosis after muscle injury and will define the SC mRNA transcriptome. Aim 1 will also determine whether Acvr1(R206H) expression in FAPs disrupts FAP-SC interactions. Aim 2 will use RNA-Seq to define the FAP transcriptome at early, critical, times after injury to identify gene targets of Acvr1(R206H) signaling. Gro-Seq (Genome-wide Run-On) analyses will quantify changes in active gene transcription, will identify candidate genes regulated by promoter-proximal polymerase pausing, and will identify potential enhancer targets of Acvr1(R206H) signaling. Lineage tracing will determine whether the abnormal muscle environment differentially affects the fate of normal and mutant FAPs. In Aim 3, DTA ablation approaches will determine whether FAPs are the sole source of osteogenic cells in muscle, information that is essential for evaluating the possible efficacy of cell-specific therapies. Activin inhibition has recently emerged as a powerful potential therapy for FOP. Aim 4 will use lineage tracing to address how Activin blockade affects proliferation, survival and developmental capacity of FAPs and SCs. RNA-Seq will define the extent to which Activin inhibition “normalizes” FAP and SC transcriptomes. The proposed research will contribute significantly to an understanding of the cells and cellular interactions responsible for HO and associated inhibition of muscle regeneration, and may lead to the development of strategies for cell-based therapies.

项目摘要异位骨化（HO），骨骼肌和相关软组织中骨骼的形成，可以受伤或疾病引起的。 HO的最极端形式表现为稀有的常染色体 - 主要的遗传疾病，纤维浮肿osscificans Progenkhiva（FOP），其中HO逐渐持续通过生活，会对健康和预期寿命产生毁灭性影响。我们开发了一种新的遗传模型 FOP基于引起疾病的BMP受体ACVR1（R206H）的条件表达。使用此模型，我们确定了纤维辅助祖细胞（FAPS），一种肌肉组织中的多能间充质祖细胞，为引起疾病的细胞种群。值得注意的是，我们已经表明，肌内注射BMP2为野生型小鼠还导致FAP衍生的HO，表明FOP患者和HO发病机理的机理相似性和普通人群。 HO的研究几乎完全集中在骨骼生长的机制上，但严重肌肉损失可能是导致患者发病率的重要因素。这个的总体目标研究是1）了解如何颠覆FAP编程以颠覆病理骨的形成； 2）了解在容易受到HO的骨骼肌中抑制再生的方式； 3）确定是否骨骼肌完全取决于FAP。通过将ACVR1R206H表达靶向使用PDGRFαCreer 和TIE2-CRE驱动程序，AIM 1将量化再生障碍，并确定肌肉干细胞是否（卫星细胞; SC）功能障碍负责再生缺陷。这些研究将量化SC增殖肌肉损伤后的凋亡，并将定义SC mRNA转录组。 AIM 1还将确定是否 FAP中的ACVR1（R206H）表达破坏了FAP-SC的相互作用。 AIM 2将使用RNA-Seq定义FAP 损伤后早期，临界时间的转录组，以鉴定ACVR1（R206H）信号传导的基因靶标。 gro-seq （全基因组跑步）分析将量化活性基因转录的变化，将识别候选基因受启动子 - 羟基聚合酶暂停的调节，并将确定 ACVR1（R206H）信号传导。谱系追踪将决定异常的肌肉环境是否有不同影响正常和突变体FAP的命运。在AIM 3中，DTA消融方法将确定FAPS是否是肌肉中成骨细胞的唯一来源，这对于评估可能的效率至关重要细胞特异性疗法。激活素抑制作用最近已成为FOP的强大潜在疗法。目标4 将使用谱系跟踪来解决激活蛋白阻滞如何影响增殖，生存和发育能力 FAPS和SC。 RNA-seq将定义激活素抑制“归一化” FAP和SC的程度转录组。拟议的研究将对细胞和细胞的理解产生重大贡献负责HO和相关抑制肌肉再生的相互作用，可能导致制定基于细胞的疗法的策略。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Overexpression of Wild-Type ACVR1 in Fibrodysplasia Ossificans Progressiva Mice Rescues Perinatal Lethality and Inhibits Heterotopic Ossification.

DOI：
10.1002/jbmr.4617
发表时间：
2022-11
期刊：
JOURNAL OF BONE AND MINERAL RESEARCH
影响因子：
6.2
作者：
Yamamoto, Masakazu;Stoessel, Sean J.;Yamamoto, Shoko;Goldhamer, David J.
通讯作者：
Goldhamer, David J.

An anti-ACVR1 antibody exacerbates heterotopic ossification by fibro-adipogenic progenitors in fibrodysplasia ossificans progressiva mice.

DOI：
10.1172/jci153795
发表时间：
2022-06-15
期刊：
JOURNAL OF CLINICAL INVESTIGATION
影响因子：
15.9
作者：
Lees-Shepard, John B.;Stoessel, Sean J.;Chandler, Julian T.;Bouchard, Keith;Bento, Patricia;Apuzzo, Lorraine N.;Devarakonda, Parvathi M.;Hunter, Jeffrey W.;Goldhamer, David J.
通讯作者：
Goldhamer, David J.