Aberrant splicing contributes to severe α-spectrin–linked congenital hemolytic anemia

Abstract

with the NMD inhibitor cycloheximide (100 μg/ml, MilliporeSigma, C1988) for 4 hours, or with a combination of both. The amount of total α-spectrin mRNA transcripts and α LEPRA mRNA transcripts were determined by real-time RT-PCR with fluorescent probes as described above. Analyses included 3 biologic and 3 technical replicates. RNA-seq data. RNA-seq data sets accessed from the Gene Expression Omnibus database were GSE61566 and GSE53983 for human erythroid cells and GSM958729 for K562 cells. Statistics. GraphPad Prism 8 software (Graph Pad Software) was used for statistical analyses. All data were reported as mean ± SD in tables or as error bars. Comparisons between 2 groups were performed using the 2-tailed Student's t test. Multiple test correction was performed using the Benjamini and Hochberg method (51). Significance was set at P less than 0.05. Human subjects. All human studies were approved by the Yale University Human Investigation Committee review board. Written informed consent was received from participants or their parents, as appropriate, prior to inclusion in the study.

Genotype‐phenotype correlation and molecular heterogeneity in pyruvate kinase deficiency

“…An example of such a case was published in which a deep intronic mutation (c.283+109C>T, intron 2) was coupled with missense mutation p.G332S . Since an increasing number of pathogenic deep intronic mutations are being described across different disease conditions (α spectrin LEPRA (SPTA1 c.4339‐99C>T) and Androgen Insensitivity Syndrome (c2450‐118A>G)), clinical molecular testing algorithms may have to be revised to include extended intronic regions, especially when only one missense mutation is identified in the first pass.…”

A family affair—Severe fetal and neonatal hemolytic anemia due to novel alpha‐spectrin mutations in two siblings

“…Truncated variants typically result in decreased alpha‐spectrin levels. The c.1677G>T VOUS may alter alpha‐spectrin splicing; altered mRNA splicing can result in defective or deficient protein products (Gallagher et al, ). It can be assumed that each mutation separately does not cause symptomatic disease, as both parents are asymptomatic.…”

Exome sequencing for diagnosis of congenital hemolytic anemia

“…Different NGS strategies have been developed over the last few years for CHA (Additional file 1, Supplementary Table S1). They included targeted gene panels (comprising 28 to 76 genes) with a success rate for diagnosis varying from 38 to 90%, depending on proband numbers (ranging from 2 to 62) and subtypes of anemia [15][16][17][18][19][20][21][22][23], and whole-exome sequencing (WES) in a few studies (limited to 1 to 7 probands, except two studies focusing on 38 Chinese cases of HS [24] and 24 cases of autosomal recessive HS [25]), with success rates between 29 and 100% [14,[24][25][26][27][28][29][30][31][32]. We used WES to explore 40 CHA patients: 20 with suspected HS and 20 with unexplained hemolysis (UH) despite available phenotype exploration.…”