Engineering a Human Skeletal Muscle Tissue Model of LGMD2B

设计 LGMD2B 的人体骨骼肌组织模型

基本信息

批准号：
10719721
负责人：
Nenad Bursac
金额：
$ 52.59万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-15 至 2028-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10719721
关键词：
3-Dimensional Adipose tissue Age Animal Model Beds Biochemical Biopsy Calcium Cell Culture Techniques Cell Separation Cell membrane Cells Cholesterol Cholesterol Homeostasis Clinical Clinical Trials Coculture Techniques Complement Complex DYSF gene Disease Disease Progression Disease model Engineering Environment Esterification Exercise Exhibits Fatty acid glycerol esters Functional disorder Generations Genes Goals Histologic Homeostasis Human Impairment In Vitro Infiltration Inflammation Inflammatory Injury Intramuscular Investigation Limb structure Limb-Girdle Muscular Dystrophies Lipids Macrophage Mediating Membrane Membrane Proteins Metabolic Mitochondria Modeling Mus Muscle Muscle Cells Muscle Fibers Muscle Weakness Muscle satellite cell Muscular Atrophy Muscular Dystrophies Mutation Myocardium Myopathy Organelles Pathogenicity Patients Phagocytosis Pharmaceutical Preparations Pharmacology Study Phenotype Play Process Proteins Regulation Reproducibility Role Severity of illness Signal Pathway Skeletal Muscle Source Study models System Testing Time Tissue Engineering Tissue Model Tissues Toxin acetyl-LDL cell motility cholesterol biosynthesis cytokine disease phenotype drug discovery dysferlinopathies experience human model immune cell infiltrate in vitro Model in vivo induced pluripotent stem cell inflammatory milieu interstitial cell lipid metabolism muscle engineering new therapeutic target novel organ growth organ on a chip pharmacologic pre-clinical repaired response response to injury therapeutic target translational study uptake

项目摘要

Limb girdle muscular dystrophy 2B (LGMD2B) is a late-onset progressive muscular dystrophy resulting from mutations in the dysferlin gene. Dysferlin is a membrane-associated protein, highly expressed in skeletal and cardiac muscle fibers where it orchestrates membrane repair in response to various injuries. Currently, there are no ongoing clinical trials or therapies to slow disease progression or cure LGMD2B. While useful for in vivo mechanistic studies, dysferlin-deficient (BLAJ) mice, a model of LGDM2B, exhibit a mild disease phenotype compared to humans, limiting mouse utility for translational studies. Developing a high-fidelity in vitro model of human LGMD2B muscle would complement mouse studies and allow patient-specific disease modeling and drug discovery. Thus, the overarching goal of this project is to engineer a novel 3D human skeletal muscle tissue model (“myobundle”) that replicates the main structural, functional, and metabolic features of LGMD2B. Specifically, we will utilize human iPSC lines from three healthy and three LGMD2B donors to engineer LGMD2B myobundles that exhibit reproducible deficits in muscle contractile function, calcium homeostasis, and lipid handling, while showing drug responses consistent with studies in BLAJ mice and LGMD2B patients. Importantly, a defining feature of LGMD2B muscle is the ectopic fat formation suggested to occur due to adipogenic differentiation of muscle interstitial cells (MICs). We will thus develop a novel tissue-engineered model of intramuscular adipose tissue (IMAT) accumulation in LGMD2B muscle by co-culturing MICs isolated from LGMD2B human muscle biopsies and iPSC-derived muscle progenitor cells. In this novel co-culture system, we will identify pro-adipogenic factors secreted from LGMD2B muscle and study their ability to induce ectopic fat formation. Since immune cell infiltration and biased macrophage polarization are additional defining features of LGMD2B muscle, we will engineer co- and tri-cultured muscle-macrophage myobundles to further characterize roles of heterocellular interactions and inflammatory milieu in injury response and fat accumulation in LGMD2B. Finally, our preliminary studies suggest that the cholesterol metabolism in LGMD2B muscle is impaired and contributes to the disease, which we will further study pharmacologically, biochemically, and histologically in LGMD2B myobundles and BLAJ mice. Overall, we expect that the novel tissue-engineered model of human LGMD2B muscle developed in this project will enable new mechanistic and pharmacological studies, eventually leading to first clinical trials for LGMD2B.

肢体肌肉营养不良2B（LGMD2B）是由疗程的晚期进行性肌营养不良症，由 Dysferlin基因中的突变。 Dysferlin是一种与膜相关的蛋白质，在骨骼和心脏肌肉纤维在响应各种损伤的情况下策划膜修复。目前，有没有正在进行的临床试验或疗法来减缓疾病进展或治愈LGMD2B。虽然对体内有用机械研究，Dysferlin缺乏症（BLAJ）小鼠，一种LGDM2B的模型，暴露了轻度疾病表型与人类相比，限制了小鼠效用进行翻译研究。开发高保真的体外模型人LGMD2B肌肉将补充小鼠研究，并允许患者特异性疾病建模和药物发现。那个项目的总体目标是设计一种新颖的3D人类骨骼肌肉组织复制LGMD2B的主要结构，功能和代谢特征的模型（“ myobundle”）。具体而言，我们将利用来自三个健康和三个LGMD2B捐赠者到工程师LGMD2B的人类IPSC线暴露可再现的肌收缩在肌肉收缩功能，钙稳态和脂质中定义处理，同时显示与BLAJ小鼠和LGMD2B患者的研究一致的药物反应。重要的是， LGMD2B肌肉的一个定义特征是建议由于脂肪形成而发生生态脂肪肌肉间质细胞（MIC）的分化。因此，我们将开发一个新型的组织工程模型通过从从中分离出来的MIC LGMD2B人类肌肉活检和IPSC衍生的肌肉祖细胞。在这个新颖的共培养系统中，我们将确定从LGMD2B肌肉分泌的促脂因素，并研究其诱导生态脂肪的能力形成。由于免疫球浸润和偏置巨噬细胞极化是其他定义特征 lgmd2b肌肉，我们将设计共同和三培养的肌肉巨噬细胞肌收获以进一步表征杂细胞相互作用和炎症环境在LGMD2B中的损伤反应和脂肪积累中的作用。最后，我们的初步研究表明，LGMD2B肌肉中的胆固醇代谢受损，并且为该疾病做出贡献，我们将在药学，生化和组织学上进一步研究 LGMD2B肌电收获和BLAJ小鼠。总体而言，我们期望人类的新型组织工程模型在该项目中开发的LGMD2B肌肉将使新的机械和药物研究最终能够导致LGMD2B的首次临床试验。