학술논문

Ultradeep Sequencing Analysis of Paroxysmal Nocturnal Hemoglobinuria Clones Detected by Flow Cytometry: PIG Mutation in Small PNH Clones
Original Article
Document Type
Academic Journal
Source
American Journal of Clinical Pathology. July 2021, Vol. 156 Issue 1, p72, 14 p.
Subject
Soliris (Medication)
Analysis
Genetic aspects
Eculizumab -- Analysis
Cloning -- Genetic aspects -- Analysis
Animal genetic engineering -- Genetic aspects -- Analysis
Gene mutation -- Analysis -- Genetic aspects
Swine -- Analysis -- Genetic aspects
Gene mutations -- Analysis -- Genetic aspects
Language
English
ISSN
0002-9173
Abstract
Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired hematopoietic stem cell disorder caused by somatic mutation of the PIGA gene. It is characterized by chronic intravascular hemolysis, thrombosis, and bone marrow [...]
Objectives: We aimed to determine whether small paroxysmal nocturnal hemoglobinuria (PNH) clones detected by flow cytometry (FCM) harbor PIG gene mutations with quantitative correlation. Methods: We analyzed 89 specimens from 63 patients whose PNH clone size was >0.1% by FCM. We performed ultradeep sequencing for the PIGA, PIGM, PIGT, and PIGX genes in these specimens. Results: A strong positive correlation between PNH clone size by FCM and variant allele frequency (VAF) of PIG gene mutation was identified (RBCs: r = 0.77, P < .001; granulocytes: r = 0.68, P < .001). Granulocyte clone size of 2.5% or greater and RBCs 0.4% or greater by FCM always harbored PIG gene mutations. Meanwhile, in patients with clone sizes of less than 2.5% in granulocytes or less than 0.4% in RBCs, PIG gene mutations were present in only 15.9% and 12.2% of cases, respectively. In addition, there was not a statistically significant positive correlation between FCM clone size and VAF or the presence or absence of a PIG mutation. Conclusions: Our results showed that in small PNH clones PIG gene mutations were present in only a small portion without significant correlation to VAF or the presence or absence of a PIG mutation. Key Words: Paroxysmal nocturnal hemoglobinuria; PIGA; PIGM; Ultradeep sequencing; Flow cytometry