Impact of Methylglyoxal on Diabetes Induced Progenitor Cell Dysfunction
Alexander Y. Li, M.S., Melanie Rodrigues, Ph.D., Marcelina G. Perez, B.S., Geoffrey C. Gurtner, M.D..
Stanford University, Stanford, CA, USA.
Reactive oxygen species (ROS) generation represents a likely mechanism by which diabetes impairs stem- and progenitor-cell function. Here, we determine the specific contribution of methylglyoxal, a highly potent ROS that increases in the presence of hyperglycemia, on progenitor cell dysfunction and impaired wound healing in diabetes.
We address methylglyoxal’s impact within transgenic murine models of glyoxalase1 (Glo1) knockdown and overexpression, effectively manipulating methylglyoxal breakdown in-vivo. Fluorescent cell sorting is employed to isolate progenitor cell populations from adipose tissue and bone marrow, which are further assayed utilizing single-cell microfluidic multiplexed RT-PCR. Unsupervised hierarchical clustering is used to highlight and quantify dysfunctional subpopulations of progenitor cells.
Glo1 knockdown mice demonstrate depletion of CD45-CD31-CD34+ ASC populations (2.74%) when compared to wild-type control mice (3.25%). However, they show greater number of ASCs compared to diabetic mice (1.14%). Similarly, Glo1 knockdown mice demonstrate fewer numbers of Lin-Sca+ckit+ hematopoietic stem cells (0.36%) when compared to wild-type mice (0.53%), and a greater number of these stem cells compared to diabetic mice (0.30%). We are currently testing both progenitor cell dysfunction in Glo1-depleted mice at a single cell level, and the reversal of such diabetic perturbations with Glo1 overexpression.
Although only partly representing the full oxidative load within a diabetic animal, methylglyoxal remains an extremely potent glycation agent capable of exerting significant negative impact on progenitor cells despite low concentrations. Our findings help address the utility of directed intervention in methylglyoxal catabolism, outlining avenues by which to explore new therapeutic targets for diabetic complications.
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