Ely E V, Kapinski A T, Paradi S G, Tang R, Guilak F, Collins K H
Department of Orthopedic Surgery, Washington University in Saint Louis, Saint Louis, MO 63110, USA.
Shriners Hospitals for Children-St. Louis, St. Louis, MO 63110, USA.
Eur Cell Mater. 2023 Jul-Dec;46:171-194. doi: 10.22203/eCM.v046a09. Epub 2023 Dec 29.
Adipose tissue is an active endocrine organ that can signal bidirectionally to many tissues and organ systems in the body. With obesity, adipose tissue can serve as a source of low-level inflammation that contributes to various co-morbidities and damage to downstream effector tissues. The ability to synthesize genetically engineered adipose tissue could have critical applications in studying adipokine signaling and the use of adipose tissue for novel therapeutic strategies. This study aimed to develop a method for non-viral adipogenic differentiation of genome-edited murine induced pluripotent stem cells (iPSCs) and to test the ability of such cells to engraft in mice . Designer adipocytes were created from iPSCs, which can be readily genetically engineered using CRISPR-Cas9 to knock out or insert individual genes of interest. As a model system for adipocyte-based drug delivery, an existing iPSC cell line that transcribes interleukin 1 receptor antagonist under the endogenous macrophage chemoattractant protein-1 promoter was tested for adipogenic capabilities under these same differentiation conditions. To understand the role of various adipocyte subtypes and their impact on health and disease, an efficient method was devised for inducing browning and whitening of Ipsc-derived adipocytes in culture. Finally, to study the downstream effects of designer adipocytes , we transplanted the designer adipocytes into fat-free lipodystrophic mice as a model system for studying adipose signaling in different models of disease or repair. This novel translational tissue engineering and regenerative medicine platform provides an innovative approach to studying the role of adipose interorgan communication in various conditions.
脂肪组织是一个活跃的内分泌器官，能够与体内许多组织和器官系统进行双向信号传递。在肥胖状态下，脂肪组织可作为低水平炎症的来源，导致各种合并症并损害下游效应组织。合成基因工程化脂肪组织的能力在研究脂肪因子信号传导以及将脂肪组织用于新型治疗策略方面可能具有关键应用。本研究旨在开发一种用于对经基因组编辑的小鼠诱导多能干细胞（iPSC）进行非病毒成脂分化的方法，并测试此类细胞在小鼠体内的植入能力。从iPSC中创建了定制脂肪细胞，利用CRISPR-Cas9可以很容易地对其进行基因工程改造，以敲除或插入感兴趣的单个基因。作为基于脂肪细胞的药物递送的模型系统，测试了一种在内源性巨噬细胞趋化蛋白-1启动子控制下转录白细胞介素1受体拮抗剂的现有iPSC细胞系在相同分化条件下的成脂能力。为了了解各种脂肪细胞亚型的作用及其对健康和疾病的影响，设计了一种在培养中诱导iPSC来源的脂肪细胞褐变和白化的有效方法。最后，为了研究定制脂肪细胞的下游效应，我们将定制脂肪细胞移植到无脂肪的脂肪营养不良小鼠体内，作为在不同疾病或修复模型中研究脂肪信号传导的模型系统。这个新颖的转化组织工程和再生医学平台为研究脂肪组织间通讯在各种情况下的作用提供了一种创新方法。
