Utilizing glycogen synthase kinase-3β as a marker for the diagnosis of graft-versus-host disease

Graft-versus-host disease (GVHD) is a deadly complication of allogeneic hematopoietic stem cell transplantation. Timely diagnosis is critical, because mortality rates for GVHD are high, increasing with disease severity. A diagnostic tool to predict GVHD before the onset of clinical symptoms could save many lives. On the cellular level, GVHD occurs when T cells from the transplant attack the tissues of the host, after perceiving them to be foreign. T-cell proliferation occurs even before clinical symptoms appear. Glycogen synthase kinase (GSK)-3β is a protein which regulates proliferation in many cell types including T-cells. GSK-3β has never been directly connected with GVHD and we applied GSK-3β as a novel marker for GVHD prediction, seeking herein to determine whether GSK-3β can be utilized as a marker for the early diagnosis of GVHD.

For the mouse model of acute GVHD, irradiated mice underwent allogeneic splenocyte transplantation and GSK-3β expression levels and phosphorylation states were monitored in harvested spleens by western blot. FACS analysis was used to measure the number of T cells within the harvested spleens.

Mice developed observable GVHD symptoms by day 5 post-transplantation, with severe symptoms on day 6 requiring mice to be killed for humane reasons. A significantly increased number of T cells in the allogeneic mice correlated with GVHD development. GSK-3β protein expression levels and phosphorylation levels were significantly lower in allogeneic (GVHD) mice compared with negative (untreated) and positive (syngeneic transplant; non-GVHD) controls over time.

GSK-3β was directly connected with the onset and progression of GVHD. Therefore, it can be utilized as a marker for GVHD diagnosis in animals and potentially in humans.