Improved diagnostic and treatment methods are needed for chronic graft-versus-host disease (cGVHD), the leading cause of late nonrelapse mortality (NRM) in long-term survivors of allogenic hematopoietic cell transplantation. Validated biomarkers that facilitate disease diagnosis and classification generally are lacking in cGVHD. Here, we conducted whole serum proteomics analysis of a well-established murine multiorgan system cGVHD model. We discovered 4 upregulated proteins during cGVHD that are targetable by genetic ablation or blocking antibodies, including the RAS and JUN kinase activator, CRKL, and CXCL7, CCL8, and CCL9 chemokines. Donor T cells lacking CRK/CRKL prevented the generation of cGVHD, germinal center reactions, and macrophage infiltration seen with wild-type T cells. Whereas antibody blockade of CCL8 or CXCL7 was ineffective in treating cGVHD, CCL9 blockade reversed cGVHD clinical manifestations, histopathological changes, and immunopathological hallmarks. Mechanistically, elevated CCL9 expression was present predominantly in vascular smooth muscle cells and uniquely seen in cGVHD mice. Plasma concentrations of CCL15, the human homolog of mouse CCL9, were elevated in a previously published cohort of 211 cGVHD patients compared with controls and associated with NRM. In a cohort of 792 patients, CCL15 measured at day 1100 could not predict cGVHD occurring within the next 3 months with clinically relevant sensitivity/specificity. Our findings demonstrate for the first time the utility of preclinical proteomics screening to identify potential new targets for cGVHD and specifically CCL15 as a diagnosis marker for cGVHD. These data warrant prospective biomarker validation studies.
Bibliographical noteFunding Information:
This work was supported by grants from the National Institutes of Health (NIH) National Cancer Institute (R01CA168814) (S.P.) (R01CA118953 and U54CA163438) (S.J.L.) (P01CA142106) (B.R.B.), NIH National Institute of Allergy and Infectious Diseases (P01 AI056299) (B.R.B.), and the NIH National Heart Lung and Blood Institute (R01HL128551) (J.K.B.); Leukemia & Lymphoma Society Scholar Award (1293-15) (S.P.); Translational Research Grant (6452-15) (B.R.B.); Lilly Physician Scientist Initiative Award (S.P.); and the Cancer Research Institute/Irvington Postdoctoral Fellowship (N.H.R.). The Chronic GVHD Consortium (U54 CA163438) is part of the National Center for Advancing Translational Science (NCATS) Rare Diseases Clinical Research Network (RDCRN). RDCRN is an initiative of the Office of Rare Disease Research, NCATS, funded through a collaboration between NCATS and the National Cancer Institute. The Fred Hutchinson Cancer Research Center Proteomics Facility is partially funded by a National Institutes of Health Cancer Center Support grant (P30 CA015704). The Fusion Orbitrap mass spectrometer was purchased with a grant from the M.J. Murdock Charitable Trust.
© 2018 by The American Society of Hematology.