Detection of NRAS mutation in cell-free DNA biological fluids from patients with kaposiform lymphangiomatosis

Orphanet Journal of Rare Diseases

  ^{1    1    2    2    3    4    4    4    4    4    5    6} 

[1] 0000 0001 2248 6943, grid.69566.3a, Department of Medical Genetics, Tohoku University School of Medicine, 980-8574, Sendai, Japan;0000 0001 2248 6943, grid.69566.3a, Division of Cell Proliferation, United Centers for Advanced Research and Translational Medicine, Tohoku University Graduate School of Medicine, 980-8575, Sendai, Japan;0000 0001 2248 6943, grid.69566.3a, Division of Interdisciplinary Medical Science, United Centers for Advanced Research and Translational Medicine, Tohoku University Graduate School of Medicine, 980-8575, Sendai, Japan;0000 0004 0370 4927, grid.256342.4, Department of Pediatrics, Graduate School of Medicine, Gifu University, Yanagido 1-1, 501-1194, Gifu, Japan;0000 0004 0373 3971, grid.136593.b, Department of Pathology, Graduate School of Medicine, Osaka University, 565-0871, Osaka, Japan;0000 0004 0467 0255, grid.415020.2, Department of Pathology, Dokkyo Medical University Saitama Medical Center, 343-8555, Saitama, Japan;

关键词: Vascular anomaly;    Kaposiform lymphangiomatosis;    Neuroblastoma RAS viral oncogene homolog;    Cell-free DNA;    Liquid biopsy;   

DOI  :  10.1186/s13023-019-1191-5

来源: publisher

【 摘 要 】

BackgroundKaposiform lymphangiomatosis (KLA) has recently been distinguished as a novel subtype of generalized lymphatic anomaly (GLA) with foci of spindle endothelial cells. All cases of KLA involve multiple organs and have an unfavorable prognosis. However, the molecular pathogenesis is unknown, and there are no useful biomarkers. In the present study, we performed genetic analysis to elucidate the cause of this disease and detect biomarkers for it.MethodsWe performed whole-exome sequencing of DNA samples from leukocytes and a biopsy specimen and analyzed cell-free DNA (cfDNA) from plasma and pleural effusion of patients to identify the NRAS c.182A > G (p.Q61R) mutation using the droplet digital polymerase chain reaction (ddPCR).ResultsAll KLA patients (patients 1–5) had invasive and aggressive features (hemorrhagic pleural effusions, coagulation disorder, and thrombocytopenia) and characteristic findings of KLA in their pathological examinations. In whole exome sequencing for patient 1, c.182A > G missense variant (p.Q61R) in NRAS was identified in fresh frozen samples of a mass on the left chest wall at a frequency of 5% of total alleles but not in his blood leukocytes. Furthermore, the same mutation was detected in cfDNA isolated from plasma and pleural effusion by using ddPCR. ddPCR analysis of plasma/pleural effusion samples from an additional four KLA patients showed that the same mutation was detected in isolated cfDNA in three of the four, as well as in a tissue sample from one of the three plasma/effusion-positive patients that had been obtained to confirm the mutation.ConclusionThese results provide the first evidence that NRAS oncogenic variant was identified in DNA samples from KLA patients from not only two affected lesions but also plasma and pleural effusion.

【 授权许可】

CC BY