PS1528 INCREASED T CELLS GLYCOLYSIS IN PATIENTS WITH ACUTE GRAFT VERSUS HOST DISEASE AFTER ALLOGENEIC HEMATOPOIETIC STEM CELL
Authors: Song Y. , Lyu Z.‐S. , Wen Q. , Yao W.‐L. , Chen Y.‐H. , Han T.‐T. , Wang Y. , Xu L.‐P. , Zhang X.‐H. , Huang X.‐J. , Kong Y. 
Published: June 2019

Background:

Acute graft‐versus‐host disease (aGVHD) is a major complication limiting successful allogeneic hematopoietic stem cell transplantation (allo‐HSCT), the pathogenesis of aGVHD is commonly considered to be T cell‐mediated. Cell metabolism determines T cell fate and function, the metabolic profile of T cells varies in different immunological disorders such as lupus, rheumatoid arthritis. Previous murine transplantation study found that alloreactive donor T cells increased both oxidative phosphorylation (OXPHOS) and aerobic glycolysis and appeared bioenergetically similar to the lymphocytes that mediate lupus. Recently, another murine study suggested that alloreactive donor T cells use glycolysis as the predominant metabolic process. However, the metabolic profile of T cells in aGVHD patients still remains unclear.

Aims:

To determine the metabolic adaptations that occur in activated T cells in patients with aGVHD and approach for modulating their metabolism.

Methods:

In this prospective case‐control study, a total of 20 patients with aGVHD after allo‐HSCT were enrolled, for each case, 1 matched transplant recipients without aGVHD (non‐aGVHD; N = 20) were selected at random. CD3+ T cells were isolated by microbeads, the metabolic profile of sorted T cells was determined using a Seahorse XF96 Analyze. RNA sequencing (RNA‐Seq) was performed to analyze the gene expression profiles of the T cells. Glucose consumption and lactate production rates were detected by glucose assay kit and lactate assay kit. The levels of intracellular cytokines, such as IFN‐r, IL‐4 and IL‐17, were detected by flow cytometry. Protein expressions for CTP1a and PFKFB3 were measured by flow cytometry. 3‐PO, as a glycolysis inhibitor at PFKFB3, was administrated to T cells from aGVHD patients.

Results:

Firstly, we detected the intracellular levels of proinflammatory cytokines (IFN‐γ, and IL‐17) in the peripheral T cells. There were significantly elevated proportions of Tc1(CD8+IFN‐γ+), Th1 (CD4+IFN‐γ+) and Th17(CD4+IL17+) in patients with aGVHD. Switching from OXPHOS to glycolysis is the hallmark of in vitro–activated T cell metabolism. Hence, we determined the rates of glycolysis and OXPHOS in T cells by measuring extracellular acidification rate (ECAR) and oxygen consumption rate (OCR). T cells isolated from aGVHD patients exhibited comparable OCR values and higher ECAR values. In addition, glucose consumption rate and lactate production rate were both higher in aGVHD group, implying higher glycolytic activity of T cells in aGVHD patients. Moreover, RNA‐seq were performed to further understand the metabolic profile of T cells in aGVHD patients. Consistent with the elevated glycolytic activity observed in vitro, the levels of the mRNAs encoding metabolic enzymes involved in the glycolytic pathway were substantially elevated in aGVHD group. Protein expression of PFKFB3, which is a glycolytic activator, was significantly higher in T cells, especially in naïve T cells of aGVHD patients. Furthermore, 3‐PO significantly diminished PFKFB3 expression, glycolytic activity of T cells, and reduced the expression of proinflammatory cytokines in T cells from aGVHD patients.

Summary/Conclusion:

In the current study, we found that T cells in patients with aGVHD preferentially depend on glycolysis to meet bioenergetic demands. Furthermore, inhibition of glycolysis through targeting PFKFB3 ameliorates the activity of T cells from aGVHD patients. Therefore, the current work indicated that glycolysis promises to be a novel therapeutic target for aGVHD patients after allo‐HSCT.