Weigand Maria, Degroote Roxane L, Amann Barbara, Renner Simone, Wolf Eckhard, Hauck Stefanie M, Deeg Cornelia A
Department of Veterinary Sciences, LMU, Munich, Germany.
Gene Center and Department of Veterinary Sciences, LMU, Munich, Germany; Center for Innovative Medical Models (CiMM), Department of Veterinary Sciences, LMU, Munich, Germany; German Center for Diabetes Research (DZD), Neuherberg, Germany.
J Proteomics. 2020 Jul 30;224:103843. doi: 10.1016/j.jprot.2020.103843. Epub 2020 May 27.
INS transgenic pigs develop a stable diabetic phenotype early after birth and therefore allow studying the influence of hyperglycemia on primary immune cells in an early stage of diabetes mellitus in vivo. Since immune response is altered in diabetes mellitus, with deviant neutrophil function discussed as one of the possible causes in humans and mouse models, we investigated these immune cells in INS transgenic pigs and wild type controls at protein level. A total of 2371 proteins were quantified by label-free LC-MS/MS. Subsequent differential proteome analysis of transgenic animals and controls revealed clear differences in protein abundances, indicating a deviant behavior of granulocytes in the diabetic state. Interestingly, abundance of myosin regulatory light chain 9 (MLC-2C) was increased 5-fold in cells of diabetic pigs. MLC-2C directly affects cell contractility by regulating myosin ATPase activity, can act as transcription factor and was also associated with inflammation. It might contribute to impaired neutrophil cell adhesion, migration and phagocytosis. Our study provides novel insights into proteome changes in neutrophils from a large animal model for permanent neonatal diabetes mellitus and points to dysregulation of neutrophil function even in an early stage of this disease. Data are available via ProteomeXchange with identifier PXD017274. SIGNIFICANCE: Our studies provide novel basic information about the neutrophil proteome of pigs and contribute to a better understanding of molecular mechanisms involved in altered immune cell function in an early stage diabetes. We demonstrate proteins that are dysregulated in neutrophils from a transgenic diabetic pig and have not been described in this context so far. The data presented here are highly relevant for veterinary medicine and have translational quality for diabetes in humans.
胰岛素转基因猪在出生后早期就会出现稳定的糖尿病表型，因此能够在体内糖尿病早期阶段研究高血糖对原代免疫细胞的影响。由于糖尿病患者的免疫反应会发生改变，在人类和小鼠模型中，中性粒细胞功能异常被认为是可能的原因之一，我们在蛋白质水平上研究了胰岛素转基因猪和野生型对照中的这些免疫细胞。通过无标记液相色谱-串联质谱法对总共2371种蛋白质进行了定量分析。随后对转基因动物和对照进行的差异蛋白质组分析揭示了蛋白质丰度的明显差异，表明糖尿病状态下粒细胞的行为异常。有趣的是，糖尿病猪细胞中肌球蛋白调节轻链9（MLC-2C）的丰度增加了5倍。MLC-2C通过调节肌球蛋白ATP酶活性直接影响细胞收缩性，可作为转录因子，并且还与炎症相关。它可能导致中性粒细胞的细胞黏附、迁移和吞噬功能受损。我们的研究为永久性新生儿糖尿病大型动物模型中性粒细胞的蛋白质组变化提供了新的见解，并指出即使在该疾病的早期阶段中性粒细胞功能也存在失调。数据可通过ProteomeXchange获得，标识符为PXD017274。意义：我们的研究提供了有关猪中性粒细胞蛋白质组的新的基础信息，并有助于更好地理解糖尿病早期免疫细胞功能改变所涉及的分子机制。我们展示了转基因糖尿病猪中性粒细胞中失调的蛋白质，这些蛋白质在此背景下迄今尚未被描述。此处呈现的数据与兽医学高度相关，并且对人类糖尿病具有转化价值。
