Cell cycle control of adipogenesis

脂肪生成的细胞周期控制

基本信息

批准号：
10476647
负责人：
Mary N Teruel
金额：
$ 21.12万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
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：
2021-09-16 至 2022-09-15
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10476647
关键词：
3-Dimensional Adipocytes Adipose tissue Adolescent Biological Assay Body Weight decreased Brown Fat CDK2 gene CDK4 gene Cell Cycle Cell Cycle Progression Cell Cycle Regulation Cell Density Cell Differentiation process Cell division Cell model Cells Child Clonal Expansion Cyclins Data Ensure Extracellular Matrix Fatty acid glycerol esters Functional disorder G1 Phase Genetic Goals Health Human Hypertrophy Image Analysis In Vitro Knock-out Knockout Mice Link Maintenance Metabolic Metabolic Diseases Methods Mitotic Molecular Mus Muscle Natural regeneration Neurons Obesity Obesity Epidemic Outcome Pancreas Pathway interactions Phase Process Proliferating Publications Publishing RHOA gene Reporter Research Risk Role SKP2 gene Signal Pathway Signal Transduction Skeletal Muscle Testing Therapeutic Time Tissue Differentiation Tissues Wild Type Mouse Work adipocyte differentiation adult obesity cellular imaging experimental study imaging approach in vivo inhibitor/antagonist interest knockout gene lipid biosynthesis live cell imaging mouse model progenitor programs regenerative tissue stem cells synergism

3-Dimensional Adipocytes Adipose tissue Adolescent Biological Assay Body Weight decreased Brown Fat CDK2 gene CDK4 gene Cell Cycle Cell Cycle Progression Cell Cycle Regulation Cell Density Cell Differentiation process Cell division Cell model Cells Child Clonal Expansion Cyclins Data Ensure Extracellular Matrix Fatty acid glycerol esters Functional disorder G1 Phase Genetic Goals Health Human Hypertrophy Image Analysis In Vitro Knock-out Knockout Mice Link Maintenance Metabolic Metabolic Diseases Methods Mitotic Molecular Mus Muscle Natural regeneration Neurons Obesity Obesity Epidemic Outcome Pancreas Pathway interactions Phase Process Proliferating Publications Publishing RHOA gene Reporter Research Risk Role SKP2 gene Signal Pathway Signal Transduction Skeletal Muscle Testing Therapeutic Time Tissue Differentiation Tissues Wild Type Mouse Work adipocyte differentiation adult obesity cellular imaging experimental study imaging approach in vivo inhibitor/antagonist interest knockout gene lipid biosynthesis live cell imaging mouse model progenitor programs regenerative tissue stem cells synergism

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项目摘要

Recent evidence shows that creating new fat cells (adipogenesis) can counteract the harmful metabolic effects of obesity, which have been shown to originate primarily from abnormal enlargement (hypertrophy) and resulting dysfunction of existing fat cells. Thus, how to increase adipogenesis over hypertrophy is of great interest. Previous work showed that preadipocytes in vivo and in vitro must complete one or more cell cycles, a process referred to as clonal expansion, before they can differentiate into adipocytes. One of the most striking genetic examples of increased adipogenesis was demonstrated by manipulating this clonal expansion period by knocking out the cell cycle inhibitors p21 and p27. Knockout of either p21 or p27 in mice results in a 2-fold increase in adipogenesis and fat mass, but knockout of both results in a dramatic 6-fold increase. Previous work suggests that p21 and p27 are regulated by very different mechanisms and are active at different times during adipogenesis. In recent published work, we identified the molecular mechanisms that could explain the increased adipogenesis observed in the p21- knockout mouse. However, how p27 regulates adipogenesis and tissue mass and how p21 and p27 synergize are poorly understood. We hypothesize that the synergistic expression of p21 and p27 during adipogenesis is the primary mechanism that controls the number of cell divisions before differentiation, and thus controls the number the adipocytes produced per preadipocyte. We will test this hypothesis by using live-cell imaging approaches. We will first use methods to inducibly express and rapidly degrade p27 to determine when and how p27 regulates cell cycle progression during adipogenesis. We will then use live cell reporters for CDK4/6 and CDK2 activity to understand when and how p27 and p21 synergize to regulate CDK4/6 and CDK2 activity and control the number of adipocytes produced. Finally, we will use live-cell reporters and p27 knockout cell and mouse models to understand how p27 is regulated by upstream cell density and extracellular matrix stiffness signals to open and close a proliferative window during adipogenesis. The outcome of this work will be a framework how the clonal expansion period can be controlled to significantly increase adipogenesis over hypertrophy while also ensuring that the progenitor pool is maintained. Our results will likely have broad applicability not only to the maintenance of adipose tissue, but also more generally for the maintenance and regeneration of neuronal, muscle, and other terminally differentiated tissues.

最近的证据表明，创建新的脂肪细胞（脂肪形成）可以抵消肥胖的有害代谢作用，已显示出主要起源于异常现有脂肪细胞的扩大（肥大）和产生的功能障碍。因此，如何增加对肥大的脂肪形成引起了极大的兴趣。先前的工作表明，前脂肪细胞体内和体外必须完成一个或多个细胞周期，这一过程称为克隆膨胀，在它们可以区分成脂肪细胞之前。最引人注目的遗传例子之一通过通过操纵克隆扩张期来证明脂肪形成的增加淘汰细胞周期抑制剂p21和p27。小鼠的p21或p27的敲除在脂肪生成和脂肪质量的增长2倍中，但两种敲除导致剧烈的6倍增加。先前的工作表明，p21和p27受到截然不同的机制的调节在成生期间的不同时间处于活跃状态。在最近发表的工作中，我们确定了可以解释p21-中观察到的脂肪形成增加的分子机制淘汰鼠标。但是，p27如何调节脂肪形成和组织质量以及p21和p27 协同作用知之甚少。我们假设p21和p27的协同表达在成生期间是控制细胞分裂数量之前的主要机制分化，从而控制每个脂肪细胞产生的脂肪细胞的数量。我们将通过使用活细胞成像方法检验该假设。我们将首先使用方法来诱导表达和快速降解p27，以确定p27何时以及如何调节细胞周期进程在脂肪生成期间。然后，我们将使用活细胞记者进行CDK4/6和CDK2活动了解P27和P21何时以及如何协同调节CDK4/6和CDK2活性以及控制产生的脂肪细胞数量。最后，我们将使用活细胞记者和P27 敲除细胞和小鼠模型，以了解p27如何受到上游细胞密度和细胞外基质刚度信号在成脂过程中打开和关闭增殖窗口。这项工作的结果将是一个框架，如何控制克隆扩展期为了显着增加肥大的脂肪形成，同时确保祖先维护池。我们的结果可能不仅适用于维护脂肪组织，但更普遍地用于维持和再生神经元，肌肉，和其他终极分化的组织。