Identification of acquired PIGA mutations and additional variants by next‐generation sequencing in paroxysmal nocturnal hemoglobinuria

Abstract

IntroductionParoxysmal Nocturnal Hemoglobinuria (PNH) is an acquired clonal disease of hematopoietic stem cells. It is caused by somatic mutation of the X‐linked PIGA gene, resulting in a deficient expression of glycosylphosphatidylinositol‐anchored proteins (GPI‐APs). In this study, we aimed to explore the diagnostic value of next‐generation sequencing (NGS) and potential molecular basis in PNH patients.MethodsGenomic DNA of 85 PNH patients was analyzed by a 114‐gene NGS panel.ResultsMutational analysis of PIGA identified 124 mutations in 92% PNH patients, including 101 distinct mutations and 23 recurrent mutations. Among them, 102 mutations were newly reported. Most mutations were located in exon 2 of PIGA gene, and truncated mutation was the most common one. Other mutations were detected in 26 out of 85 cases, including five cases of DNMT3A variants, four cases of ASXL1 variants, and four cases of U2AF1 variants. Clonal analysis was performed in one case and outlined a linear evolution pattern in classic PNH. There was a positive correlation between number of PIGA mutations and fraction of GPI‐APs deficient granulocytes.ConclusionThe detection of PIGA mutations and additional variants by targeted NGS not only shed light on the genetic characteristics of PNH, but also provided an important reference value in the diagnosis of PNH at molecular level.

GPI-AP: Unraveling a New Class of Malignancy Mediators and Potential Immunotherapy Targets

“…Furthermore, two subsets of mutations were associated with early infantile epileptic-dyskinetic encephalopathy; first is the homozygous c.384del variant of PIG-P gene (25), which led to the frame shift of 6 codons before the expected stop signal (25), and second are PIG-Q mutations, particularly PIG-Q novel variants, which included two missense mutations (p.G17R; p.G449R), a canonical splice site substitution (c.942+1G>A), an in-frame deletion (p.A377_S389del) and three frameshifts (p.Q527Afs*75, p.R538Afs*24 and p.G557Dfs*) (26). Finally, mutational analysis of PIGA identified 124 PIG-A mutations in 92% of paroxymal nocturnal hemoglobinuria (PNH) patients, of which 101 were distinct mutations and 23 were recurrent (27).…”

“…Some studies tested the impact of GPI pathway manipulation on dependent GPI-anchored complement regulatory protein, CD-59, which was found to be under expressed in congenital and neurological disorders, as well as PNH (21)(22)(23)(24)(25)(26)(27). CD-59 was, however, overexpressed in most solid tumors (28).…”

Transcriptome analysis reveals sexual disparities in gene expression in rat brain microvessels

“…Directly relating to brain MVs, GPI-APs have been reported to regulate leukocyte adhesion and thereby protect endothelium from damage. 23 Defects in the GPI-anchor synthesis occur in paroxysmal nocturnal hemohemoglobinuria 24 and congenital diseases. 25 Reck, a GPI-AP, plays an important role in neurovascular development.…”

“…Directly relating to brain MVs, GPI-APs have been reported to regulate leukocyte adhesion and thereby protect endothelium from damage. 23 Defects in the GPI-anchor synthesis occur in paroxysmal nocturnal hemohemoglobinuria 24 and congenital diseases. 25 Reck, a GPI-AP, plays an The results for statistical significance of (e) using merged western blot data were identical whether we used the Student's t-test or two-way analysis of variance.…”

Coexistence of Primary Myelofibrosis and Paroxysmal Nocturnal Hemoglobinuria Clone with JAK2 V617F, U2AF1 and SETBP1 Mutations: A Case Report and Brief Review of Literature

“…Furthermore, the application of NGS in PNH patients demonstrated that the mutations of myeloid neoplasm-related genes (i.e., TET2 , SUZ12 , U2AF1 , JAK2 , ASXL1 , DNMT3A , etc.) could be acquired in addition to the PIGA gene mutation [ 8 , 9 , 10 ].…”