Lavoie Jessie R, Creskey Marybeth M, Muradia Gauri, Bell Gillian I, Sherman Stephen E, Gao Jun, Stewart Duncan J, Cyr Terry D, Hess David A, Rosu-Myles Michael
Regulatory Research Division, Centre for Biologics Evaluation, Biologics and Genetic Therapies Directorate, Health Products and Food Branch, Health Canada, Ottawa, Ontario, Canada.
Department of Physiology and Pharmacology, Molecular Medicine Research Group, Krembil Centre for Stem Cell Biology, Robarts Research Institute, University of Western Ontario, London, Ontario, Canada.
Stem Cells. 2016 Aug;34(8):2249-55. doi: 10.1002/stem.2385. Epub 2016 May 19.
Multipotent mesenchymal stromal cell (MSC) transplantation is proposed as a novel therapy for treating diabetes by promoting the regeneration of damaged islets. The clinical promise of such treatments may be hampered by a high degree of donor-related variability in MSC function and a lack of standards for comparing potency. Here, we set out to identify markers of cultured human MSCs directly associated with islet regenerative function. Stromal cultures from nine separate bone marrow donors were demonstrated to have differing capacities to reduce hyperglycemia in the NOD/SCID streptozotocin-induced diabetic model. Regenerative (R) and non-regenerative (NR) MSC cultures were directly compared using isobaric tags for relative and absolute quantitation (iTRAQ)-based quantitative proteomics. A total of 1,410 proteins were quantified resulting in the identification of 612 upregulated proteins and 275 downregulated proteins by ± 1.2-fold in R-MSC cultures. Elastin microfibril interface 1 (EMILIN-1), integrin-linked protein kinase (ILK), and hepatoma-derived growth factor (HDGF) were differentially expressed in R-MSCs, and Ingenuity Pathway Analyses revealed each candidate as known regulators of integrin signaling. Western blot validation of EMILIN-1, ILK, and HDGF not only showed significantly higher abundance levels in R-MSCs, as compared with NR-MSCs, but also correlated with passage-induced loss of islet-regenerative potential. Generalized estimating equation modeling was applied to examine the association between each marker and blood glucose reduction. Both EMILIN-1 and ILK were significantly associated with blood glucose lowering function in vivo. Our study is the first to identify EMILIN-1 and ILK as prospective markers of islet regenerative function in human MSCs. Stem Cells 2016;34:2249-2255.
多能间充质基质细胞（MSC）移植被认为是一种通过促进受损胰岛再生来治疗糖尿病的新疗法。此类治疗的临床前景可能会受到MSC功能中高度的供体相关变异性以及缺乏效力比较标准的阻碍。在此，我们着手鉴定与胰岛再生功能直接相关的培养人MSC的标志物。在NOD/SCID链脲佐菌素诱导的糖尿病模型中，来自9个不同骨髓供体的基质培养物显示出不同的降低高血糖的能力。使用基于等压标签相对和绝对定量（iTRAQ）的定量蛋白质组学直接比较再生（R）和非再生（NR）MSC培养物。共定量了1410种蛋白质，结果在R-MSC培养物中鉴定出612种上调蛋白质和275种下调蛋白质，其变化幅度为±1.2倍。弹性蛋白微原纤维界面1（EMILIN-1）、整合素连接蛋白激酶（ILK）和肝癌衍生生长因子（HDGF）在R-MSC中差异表达， Ingenuity通路分析显示每个候选物都是整合素信号的已知调节因子。EMILIN-1、ILK和HDGF的蛋白质印迹验证不仅显示与NR-MSC相比，R-MSC中的丰度水平显著更高，而且与传代诱导的胰岛再生潜力丧失相关。应用广义估计方程模型来检验每个标志物与血糖降低之间的关联。EMILIN-1和ILK在体内均与血糖降低功能显著相关。我们的研究首次将EMILIN-1和ILK鉴定为人MSC中胰岛再生功能的前瞻性标志物。《干细胞》2016年；34：2249 - 2255。
