Molecular and clinical profile of VWD in a large cohort of Chinese population: application of next generation sequencing and CNVplex® technique

SummaryVon Willebrand disease (VWD), the most common inherited bleeding disorder, is characterised by a variable bleeding tendency, heterogeneous laboratory phenotype and race specific distribution of mutations. The present study aimed to determine the correlation of genotype and phenotype in 200 Chinese individuals from 90 unrelated families with VWD. Next generation sequencing (NGS) of the whole coding VWF, copy number analysis of VWF by CNVplex® technique as well as a comprehensive phenotypic assessment were carried out in all index patients (IPs). We identified putative mutations in all IPs except five mild type 1 (85/90, 94.4%). In total, 98 different mutations were detected, 62 (63.3% of which were reported for the first time (23 missense mutations, 1 regulatory mutation, 12 splice site mutations and 26 null mutations). Mutations p.Ser1506Leu and p.Arg1374His/Cys/ Ser were the most frequent mutations in 2A (33% of cases) and 2M VWD (67% of cases), respectively. In addition, mutation p.Arg816Trp was detected repeatedly in type 2N patients, while mutation p.Arg854Gln, extremely common in Caucasians, was not found in our cohort. Thirty-three patients had two or more putative mutations. Unlike most cases of type 1 and type 2 VWD, which were transmitted dominantly, we presented seven severe type 1, two type 2A and one type 2M with autosomal recessive inheritance. Here the phenotypic data of patients with novel mutations will certainly contribute to the better understanding of the molecular genetics of VWF-related phenotypes.Supplementary Material to this article is available online at www.thrombosis-online.com.

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Next-Generation Sequencing Reveals ThatHLA-DRB3,-DRB4, and-DRB5May Be Associated With Islet Autoantibodies and Risk for Childhood Type 1 Diabetes

Diabetes ◽

10.2337/db15-1115 ◽

2016 ◽

Vol 65 (3) ◽

pp. 710-718 ◽

Cited By ~ 32

Author(s):

Lue Ping Zhao ◽

Shehab Alshiekh ◽

Michael Zhao ◽

Annelie Carlsson ◽

Helena Elding Larsson ◽

...

Keyword(s):

Type 1 Diabetes ◽

Next Generation Sequencing ◽

Islet Autoantibodies ◽

Next Generation ◽

Childhood Type 1 Diabetes ◽

Childhood Type ◽

Generation Sequencing

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Resolving Differential Diagnostic Problems in von Willebrand Disease, in Fibrinogen Disorders, in Prekallikrein Deficiency and in Hereditary Hemorrhagic Telangiectasia by Next-Generation Sequencing

Life ◽

10.3390/life11030202 ◽

2021 ◽

Vol 11 (3) ◽

pp. 202

Author(s):

Réka Gindele ◽

Adrienne Kerényi ◽

Judit Kállai ◽

György Pfliegler ◽

Ágota Schlammadinger ◽

...

Keyword(s):

Next Generation Sequencing ◽

Hereditary Hemorrhagic Telangiectasia ◽

Von Willebrand Disease ◽

Differential Diagnostics ◽

Bleeding Disorders ◽

Next Generation ◽

Illumina Platform ◽

Clinical Patient ◽

Von Willebrand ◽

Generation Sequencing

Diagnosis of rare bleeding disorders is challenging and there are several differential diagnostics issues. Next-generation sequencing (NGS) is a useful tool to overcome these problems. The aim of this study was to demonstrate the usefulness of molecular genetic investigations by summarizing the diagnostic work on cases with certain bleeding disorders. Here we report only those, in whom NGS was indicated due to uncertainty of diagnosis or if genetic confirmation of initial diagnosis was required. Based on clinical and/or laboratory suspicion of von Willebrand disease (vWD, n = 63), hypo-or dysfibrinogenemia (n = 27), hereditary hemorrhagic telangiectasia (HHT, n = 10) and unexplained activated partial thromboplastin time (APTT) prolongation (n = 1), NGS using Illumina platform was performed. Gene panel covered 14 genes (ACVRL1, ENG, MADH4, GDF2, RASA1, F5, F8, FGA, FGB, FGG, KLKB1, ADAMTS13, GP1BA and VWF) selected on the basis of laboratory results. We identified forty-seven mutations, n = 29 (6 novel) in vWD, n = 4 mutations leading to hemophilia A, n = 10 (2 novel) in fibrinogen disorders, n = 2 novel mutations in HHT phenotype and two mutations (1 novel) leading to prekallikrein deficiency. By reporting well-characterized cases using standardized, advanced laboratory methods we add new pieces of data to the continuously developing “bleeding disorders databases”, which are excellent supports for clinical patient management.

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Discovery of mutations in Chenopodium quinoa Willd through EMS mutagenesis and mutation screening using pre-selection phenotypic data and next-generation sequencing

The Journal of Agricultural Science ◽

10.1017/s0021859619000182 ◽

2018 ◽

Vol 156 (10) ◽

pp. 1196-1204 ◽

Cited By ~ 1

Author(s):

Camilo Mestanza ◽

Ricardo Riegel ◽

Santiago C. Vásquez ◽

Diana Veliz ◽

Nicolás Cruz-Rosero ◽

...

Keyword(s):

Next Generation Sequencing ◽

Mutation Screening ◽

Chenopodium Quinoa ◽

Acetolactate Synthase ◽

Screening Strategy ◽

Next Generation ◽

Induced Mutations ◽

Phenotypic Data ◽

Next Generation Sequencing Ngs ◽

Generation Sequencing

AbstractQuinoa (Chenopodium quinoaWilld) is a dicotyledonous annual species belonging to the family Amaranthaceae, which is nutritionally well balanced in terms of its oil, protein and carbohydrate content. Targeting-induced local lesions in genomes (the TILLING strategy) was employed to find mutations in acetolactate synthase (AHAS) genes in a mutant quinoa population. TheAHASgenes were targeted because they are common enzyme target sites for five herbicide groups. Ethyl methane sulfonate (EMS) was used to induce mutations in theAHASgenes; it was found that 2% EMS allowed a mutation frequency of one mutation every 203 kilobases to be established. In the mutant population created, a screening strategy using pre-selection phenotypic data and next-generation sequencing (NGS) allowed identification of a mutation that alters the amino acid composition of this species (nucleotide 1231 codon GTT→ATT, Val→Ile); however, this mutation did not result in herbicide resistance. The current work shows that TILLING combined with the high-throughput of NGS technologies and an overlapping pool design provides an efficient and economical method for detecting induced mutations in pools of individuals.

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Prenatal Diagnosis of Tyrosinemia Type 1 Using Next Generation Sequencing

Fetal and Pediatric Pathology ◽

10.3109/15513815.2016.1167149 ◽

2016 ◽

Vol 35 (4) ◽

pp. 282-285 ◽

Cited By ~ 3

Author(s):

Maryam Rafati ◽

Faezeh Mohamadhashem ◽

Azadeh Hoseini ◽

Somayeh Darzi Ramandi ◽

Saeed Reza Ghaffari

Keyword(s):

Prenatal Diagnosis ◽

Next Generation Sequencing ◽

Next Generation ◽

Tyrosinemia Type 1 ◽

Generation Sequencing

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Targeted next generation sequencing identified a novel mutation in MYO7A causing Usher syndrome type 1 in an Iranian consanguineous pedigree

International Journal of Pediatric Otorhinolaryngology ◽

10.1016/j.ijporl.2017.10.022 ◽

2018 ◽

Vol 104 ◽

pp. 10-13 ◽

Cited By ~ 4

Author(s):

Daniz Kooshavar ◽

Masoumeh Razipour ◽

Morteza Movasat ◽

Mohammad Keramatipour

Keyword(s):

Next Generation Sequencing ◽

Novel Mutation ◽

Usher Syndrome ◽

Syndrome Type ◽

Next Generation ◽

Targeted Next Generation Sequencing ◽

Generation Sequencing ◽

Usher Syndrome Type

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Dehydrated hereditary stomatocytosis with new missense mutations in PIEZO1 through the use of next-generation sequencing panel

10.22541/au.163955457.78063581/v1 ◽

2021 ◽

Author(s):

Sultan Aydin Koker ◽

Tuba Karapınar ◽

Paola BIANCHI ◽

Yeşim Oymak ◽

Elisa Fermo ◽

...

Keyword(s):

Next Generation Sequencing ◽

Missense Mutation ◽

Hemolytic Anemia ◽

Male Patient ◽

Autoimmune Hemolytic Anemia ◽

Missense Mutations ◽

Next Generation ◽

Targeted Next Generation Sequencing ◽

Generation Sequencing

In this case study, we report an 11-year-old male patient who had jaundice, hepatosplenomegaly, and chronic mild congenital non-autoimmune hemolytic anemia. In our patient, a novel homozygous missense mutation in the PIEZO1 gene was detected using a gene-targeted Next-Generation Sequencing panel: c.3364G>A (p.Glu1122Lys), confirming the diagnosis of DHS.

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Novel SCNN1A Gene Splicing-site Mutation Causing Autosomal Recessive Pseudohypoaldosteronism type 1 (PHA1) in two Italian Patients Belonging to the Same Small Town

10.21203/rs.3.rs-432720/v1 ◽

2021 ◽

Author(s):

Gregorio Serra ◽

Vincenzo Antona ◽

Maria Michela D’Alessandro ◽

Maria Cristina Maggio ◽

Vincenzo Verde ◽

...

Keyword(s):

Metabolic Acidosis ◽

Next Generation Sequencing ◽

Family History ◽

Gene Mutation ◽

Small Town ◽

Next Generation ◽

Next Generation Sequencing Ngs ◽

Generation Sequencing ◽

Pseudohypoaldosteronism Type

Abstract IntroductionPseudohypoaldosteronism type 1 (PHA1) is a rare genetic disease due to the peripheral resistance to aldosterone. Its clinical spectrum includes neonatal salt loss syndrome with hyponatremia and hypochloraemia, hyperkalemia, metabolic acidosis and increased plasmatic levels of aldosterone. Two genetically distinct forms of disease, renal and systemic, have been described, showing a wide clinical expressivity. Mutations in the genes encoding for the subunits of the epithelial sodium channels (ENaC) are responsible for generalized PHA1. Patients’ presentationWe hereby report on two Italian patients with generalized PHA1, coming from the same small town in the center of Sicily. The first patient is a male child, born from the first pregnancy of healthy consanguineous Sicilian parents. A novel SCNN1A (sodium channel epithelial subunit alpha) gene mutation, inherited from both heterozygous parents, was identified by next generation sequencing (NGS) in the homozygous child (and later, also in the heterozygous maternal aunt). A more detailed family history disclosed a possible related twenty-year-old girl, belonging to the same Sicilian small town, with referred neonatal salt loss syndrome associated to hyperkalemia, and subsequent normal growth and neurodevelopment. This second patient had a PHA1 clinical diagnosis when she was about one year old. The genetic investigation was, then, extended to her and to her family, revealing the same mutation in the homozygous girl and in the heterozygous parents.ConclusionsThe neonatologist should consider PHA1 diagnosis in newborns showing hyponatremia, hyperkalemia and metabolic acidosis, after the exclusion of a salting-loss form of adrenogenital syndrome. The increased plasmatic levels of aldosterone and aldosterone/renin ratio, associated to a poor response to steroid administration, confirmed the diagnosis in the first present patient. An accurate family history may be decisive to identify the clinical picture. A multidisciplinary approach and close follow-up evaluations are requested, in view of optimal management, adequate growth and development of patients. Next generation sequencing (NGS) techniques allowed the identification of the SCNN1A gene mutation either in both patients or in other heterozygous family members, enabling also primary prevention of disease. Our report may broaden the knowledge of the genetic and molecular bases of PHA1, improving its clinical characterization and providing useful indications for the treatment of patients. Clinical approach must be personalized, also in relation to long-term survival and potential multiorgan complications.

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Next-generation sequencing reveals novel mutations in a collision tumor of glioblastoma and meningioma

CNS Oncology ◽

10.2217/cns-2020-0029 ◽

2021 ◽

pp. CNS70

Author(s):

Kelly Chamberlin ◽

Gregory Chamberlin ◽

Katherine Saunders ◽

Simon Khagi

Keyword(s):

Next Generation Sequencing ◽

Tumor Resection ◽

Altered Mental Status ◽

Collision Tumor ◽

Genetic Alterations ◽

Homozygous Deletion ◽

Missense Mutations ◽

Next Generation ◽

Collision Tumors ◽

Generation Sequencing

Primary intracranial collision tumors are rare in patients without predisposing factors. We report such a case in a 42-year-old female who presented with headaches and altered mental status. Imaging revealed a single heterogeneous, rim-enhancing lesion in the left parieto-occipital periventricular region, involving the corpus callosum. Stereotactic biopsy demonstrated glioblastoma. Subsequent tumor resection showed histologic evidence of glioblastoma and meningioma. Next-generation sequencing was performed on both tumor components. The glioblastoma exhibited a CDKN2A homozygous deletion and novel missense mutations in TAF1L and CSMD3, while no definitive genetic alterations were identified in the meningioma. Next-generation sequencing may yield insight into molecular drivers of intracranial collision tumors and aid in identifying future therapeutic targets.

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New Genotypes and Phenotypes in Patients with 3 Subtypes of Waardenburg Syndrome Identified by Diagnostic Next-Generation Sequencing

Neural Plasticity ◽

10.1155/2019/7143458 ◽

2019 ◽

Vol 2019 ◽

pp. 1-12 ◽

Cited By ~ 4

Author(s):

Wu Li ◽

Lingyun Mei ◽

Hongsheng Chen ◽

Xinzhang Cai ◽

Yalan Liu ◽

...

Keyword(s):

Next Generation Sequencing ◽

De Novo ◽

Waardenburg Syndrome ◽

Next Generation ◽

Single Nucleotide Variants ◽

Number Variation ◽

First Time ◽

Generation Sequencing

Background. Waardenburg syndrome (WS) is one of the most common forms of syndromic deafness with heterogeneity of loci and alleles and variable expressivity of clinical features. Methods. The technology of single-nucleotide variants (SNV) and copy number variation (CNV) detection was developed to investigate the genotype spectrum of WS in a Chinese population. Results. Ninety WS patients and 24 additional family members were recruited for the study. Fourteen mutations had not been previously reported, including c.808C>G, c.117C>A, c.152T>G, c.803G>T, c.793-3T >G, and c.801delT on PAX3; c.642_650delAAG on MITF; c.122G>T and c.127C>T on SOX10; c.230C>G and c.365C>T on SNAI2; and c.481A>G, c.1018C>G, and c.1015C>T on EDNRB. Three CNVs were de novo and first reported in our study. Five EDNRB variants were associated with WS type 1 in the heterozygous state for the first time, with a detection rate of 22.2%. Freckles occur only in WS type 2. Yellow hair, amblyopia, congenital ptosis, narrow palpebral fissures, and pigmentation spots are rare and unique symptoms in WS patients from China. Conclusions. EDNRB should be considered as another prevalent pathogenic gene in WS type 1. Our study expanded the genotype and phenotype spectrum of WS, and diagnostic next-generation sequencing is promising for WS.

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