PB1422 - Differential expression of proteins from cultured endothelial cells exposed to serum from patients with acute graft versus host disease sensitive or refractory to corticosteroid treatment

Tuesday, June 27, 2023

18:30 – 19:30 ET

Room: Exhibition

Presenting Author(s)

Julia Martinez-Sanchez, PhD

PhD student
Hemostasis and Erythropathology Laboratory, Hematopathology, Department of Pathology, Hospital Clínic de Barcelona, Universitat de Barcelona, Barcelona, Spain.
Barcelona, Catalonia, Spain

Co-Author(s)

MP

Marta Palomo

PhD
Hematology External Quality Assessment Laboratory, Biomedical Diagnostic Center, Hospital Clinic of Barcelona, Barcelona, Spain.
Barcelona, Catalonia, Spain
AP

Alexandra Pedraza

Medical Doctor
Blood Bank Department, Hematopoietic Transplantation Unit, Banc de Sang i Teixits, Hospital Clínic, Barcelona, Spain.
Barcelona, Catalonia, Spain
Ana Belén Moreno-Castaño, MD

Medical Doctor
Hemostasis and Erythropathology Laboratory, Hematopathology, Department of Pathology, Centre de Diagnòstic Biomèdic (CDB), Hospital Clínic de Barcelona, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Universitat de Barcelona, Barcelona, Spain.
Barcelona, Catalonia, Spain
ST

Sergi Torramade-Moix

PhD
Hemostasis and Erythropathology Laboratory, Hematopathology, Department of Pathology, Centre de Diagnòstic Biomèdic (CDB), Hospital Clínic de Barcelona, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Universitat de Barcelona, Barcelona, Spain.
Barcelona, Catalonia, Spain
MR

Montserrat Rovira

Medical Doctor
Hematology Department, Bone Marrow Transplantation Unit, Institut Clínic de Malalties Hemato-Oncològiques (ICMHO), Hospital Clínic, Barcelona, Spain.
Barcelona, Catalonia, Spain
MS

María Queralt Salas

Medical Doctor
Hematology Department, Bone Marrow Transplantation Unit, Institut Clínic de Malalties Hemato-Oncològiques (ICMHO), Hospital Clínic, Barcelona, Spain.
Barcelona, Catalonia, Spain
JC

Joan Cid

Medical Doctor
Apheresis & Cellular Therapy Unit, Department of Hemotherapy and Hemostasis, ICMHO, Hospital Clínic de Barcelona, IDIBAPS, Universitat de Barcelona, Barcelona, Spain.
Barcelona, Catalonia, Spain
GE

Gines Escolar

MD PhD Senior Consultant
Hemostasis and Erythropathology Laboratory, Hematopathology, Department of Pathology, Centre de Diagnòstic Biomèdic (CDB), Hospital Clínic de Barcelona, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Universitat de Barcelona, Barcelona, Spain.
Barcelona, Catalonia, Spain
OP

Olaf Penack

Medical Doctor
Hematology, Oncology and Tumorimmunology Department, Charité-Universitätsmedizin Berlin, Freie Universität Berlin and Humboldt-Universität zu Berlin, Berlin, Germany.
Berlin, Berlin, Germany
EC

Enric Carreras

Medical Doctor
Josep Carreras Leukaemia Research Institute, Hospital Clínic de Barcelona, Universitat de Barcelona, Barcelona, Spain.
Barcelona, Catalonia, Spain
MD

Maribel Diaz-Ricart

PhD Senior Consultant
Hemostasis and Erythropathology Laboratory, Hematopathology, Department of Pathology, Centre de Diagnòstic Biomèdic (CDB), Hospital Clínic de Barcelona, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Universitat de Barcelona, Barcelona, Spain.
Barcelona, Catalonia, Spain

Background: Graft-versus-host disease (GVHD) is a complication with high morbidity and mortality that occurs after allogeneic hematopoietic stem cell transplantation. Endothelial injury plays a crucial role as a pathophysiological substrate for GVHD. The first-line treatment is high dose corticosteroids, although some patients are steroid refractory.

Aims: We performed a comparative analysis of the endothelial proteomic profile in response to serum from patients with acute GVHD (aGVHD) sensitive and refractory to steroids.

Methods: Blood samples from aGVHD patients, sensitive (SS-GVHD, n=8) and refractory (SR-GVHD, n=4) to steroids, were collected at aGVHD diagnosis and before treatment initiation. Endothelial cell cultures (HMEC-1) were exposed to patients serum, for 48h. Cells were lysed and samples were analyzed by nanoliquid chromatography coupled to mass spectrometry, followed by isobaric labeling for protein identification and quantification. Comparisons were performed (Mass Profiler Professional) considering a p< 0.05 and 1.15-fold, and differences were statistically evaluated by multivariate analysis.

Results: Forty-four proteins contributed to separate all samples into the two study groups, according to the partial least squares discriminant analysis scores plot. Fifteen proteins participated more significantly (p < 0.05) in separating both groups, and 10 of them exhibited a fold change >1.2. Proteins differentially expressed were mainly associated with oxidative phosphorylation, such as Cytochrome C oxidase subunit 6B1 (CX6B1), and inflammation and angiogenesis, such as Apolipoprotein D (APOD), both downregulated in SR-GVHD.

Conclusion(s): This pilot study used a novel approach to distinguish the aGVHD response to steroid treatment. The results obtained represent a first step to investigate the endothelium contribution in the treatment response of those patients who develop aGVHD. The proteins differentially expressed could constitute potential biomarkers for steroid treatment response. These findings signify a step forward to identify the mechanisms of response to steroids, of high clinical relevance considering the SR-GVHD elevated mortality.