Development of an ex vivo system to assess murine adipocyte differentiation

在健康的情况下，我们身体的脂肪细胞是储存能量的重要储存库，它们还可以释放激素，影响我们吃多少，以及我们身体中不同细胞如何储存营养。然而，在肥胖的发展过程中，这些脂肪细胞储存多余脂肪的能力受到损害，脂肪细胞释放的因子类型也受到损害。这导致我们正常的血糖水平调节出现严重问题，并增加了患其他健康问题的风险，如2型糖尿病和心血管疾病。因此，需要有效的长期疗法来促进减肥。已经确定了影响脂肪细胞生长和发育的各种蛋白质。其中一个因子是14-3-3 -3zeta，这是一种独特的蛋白质，可以帮助指导蛋白质在细胞中的去向。宿主实验室之前发现，当14-3-3 -3zeta被删除时，这会阻碍脂肪细胞的生长，但出乎意料的是，他们最近发现，减少脂肪细胞中的14-3-3 -3zeta水平会导致它们失去身份，恢复到不成熟状态。虽然最初认为14-3-3 -3zeta在每个细胞中都表达，但事实并非如此，尤其是在小鼠的脂肪组织中。这表明，如果14-3-3 -3zeta水平降低，一组特定的脂肪细胞将失去其成熟度，而这是否会对整体代谢健康产生影响尚不清楚。这将在当前的项目中得到解决，该项目将使用表达荧光蛋白的转基因小鼠，该荧光蛋白允许从脂肪组织中纯化14-3-3 - 3泽塔阳性和14-3-3 - 3泽塔阴性的脂肪细胞，并将确定这些不同组的脂肪细胞是否表现不同。我们将测量各种基因的表达，并探索细胞是否在功能上有所不同。当脂肪细胞从脂肪组织中移除时，它们不能长时间存活，所以我们也将开发一种新方法来促进这些不同脂肪细胞群的长期培养，以提高我们正确研究它们的能力。这项研究的好处是双重的：从短期来看，它将提高我们对脂肪细胞生物学复杂性的理解，从长远来看，这一知识的进步可能会导致治疗肥胖症的新方法的发展，肥胖症是一种在全球范围内持续增加的慢性疾病。

MRC : Tanyel Ashik : MR/N013700/1Under healthy conditions, our body's fat cells act as important reservoirs to store energy, and they can also release hormones that influence how much we eat and how different cells in our body store nutrients. However, during the development of obesity, the capacity for these fat cells to store excess fat becomes impaired, as does the types of factors released by fat cells. This leads to profound problems with our normal regulations of blood sugar levels and increases the risk for developing other health conditions such as type 2 diabetes and cardiovascular diseases. There is therefore a need for effective long-term therapies to promote weight loss.Various proteins that influence the growth and development of fat cells have been identified. One such factor is 14-3-3zeta, a unique protein that can help to direct where proteins go in a cell. The host lab previously found that when 14-3-3zeta is deleted, this blocked the growth of fat cells, but unexpectedly, they have recently found that reducing 14-3-3zeta levels in fat cells causes them to lose their identity and revert to an immature state. While it was initially thought that 14-3-3zeta is expressed in every cell, this is not the case, especially in adipose tissue of mice. This suggests that a specific group of fat cells will lose their maturity if 14-3-3zeta levels are reduced, and whether this will have implications on overall metabolic health is not known. This will be addressed in the current project, which will use genetically modified mice expressing fluorescent proteins that allow purification of 14-3-3zeta-positive and 14-3-3zeta-negative fat cells from adipose tissues, and it will be determined if these different groups of fat cells behave differently. We will measure the expression of various genes and explore if the cells are functionally different. When fat cells are removed from adipose tissue, they cannot survive for extended periods of time so we will also develop a new method to promote the long-term culture of these different populations of fat cells to increase our ability to properly study them.The benefits of this research are two-fold: in the short term it will improve our understanding of the complexity of fat cell biology and in the longer term this advancement in knowledge may lead to the development of new approaches to treat obesity, a chronic disease that continues to increase in worldwide prevalence.