Preimplantation genetic diagnosis of hemophilia A

Chen, Ming; Chang, Shun‐Ping; Ma, Gwo‐Chin; Lin, Wen‐Hsiang; Chen, Shee‐Uan; Tsai, Horng‐Der; Tsai, Feng-Po; Shen, Ming‐Ching

doi:10.1186/s12959-016-0098-9

Cited by 13 publications

(10 citation statements)

References 41 publications

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“…Therefore, this new panel improves the odds of identifying markers that are informative for use in standalone indirect linkage analysis, or as a complement to direct mutation detection, in couples wishing to undergo PGD for HEMA. Several reports have demonstrated that multiplex marker PCR is a reliable indirect linkage-based PGD method for cases involving inversion mutations [19][20][21][22]. Hence, this tetradecaplex marker panel, which contains more markers than previously reported panels, is likely to benefit carriers of the intron-1 or intron-22-mediated inversion mutations the most, because direct single-cell mutation detection is not possible and indirect linkage analysis is the only option [19].…”

Section: Discussionmentioning

confidence: 99%

“…Microsatellite markers, also referred to as short tandem repeats (STRs), have been widely used in linkage analysis as they are highly polymorphic and widespread within the human genome. A list of STRs located in and around the F8 gene has been reported previously [10], some of which have been multiplexed in small panels [9,[11][12][13][14][15][16][17][18][19][20][21][22]. However, a majority of these STRs have not been tested experimentally and their heterozygosity and polymorphism indices are unknown.…”

Section: Introductionmentioning

confidence: 99%

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Single‐tube tetradecaplex panel of highly polymorphic microsatellite markers < 1 Mb from F8 for simplified preimplantation genetic diagnosis of hemophilia A

Zhao

Chen

Tan

et al. 2017

Journal of Thrombosis and Haemostasis

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Background It is currently not possible to perform single-cell preimplantation genetic diagnosis (PGD) to directly detect the common inversion mutations of the factor VIII (F8) gene responsible for severe hemophilia A (HEMA). As such, PGD for such inversion carriers relies on indirect analysis of linked polymorphic markers. Objectives To simplify linkage-based PGD of HEMA, we aimed to develop a panel of highly polymorphic microsatellite markers located near the F8 gene that could be simultaneously genotyped in a multiplex-PCR reaction. Methods We assessed the polymorphism of various microsatellite markers located ≤ 1 Mb from F8 in 177 female subjects. Highly polymorphic markers were selected for co-amplification with the AMELX/Y indel dimorphism in a single-tube reaction. Results Thirteen microsatellite markers located within 0.6 Mb of F8 were successfully co-amplified with AMELX/Y in a single-tube reaction. Observed heterozygosities of component markers ranged from 0.43 to 0.84, and ∼70-80% of individuals were heterozygous for ≥ 5 markers. The tetradecaplex panel successfully identified fully informative markers in a couple interested in PGD for HEMA because of an intragenic F8 point mutation, with haplotype phasing established through a carrier daughter. In-vitro fertilization (IVF)-PGD involved single-tube co-amplification of fully informative markers with AMELX/Y and the mutation-containing F8 amplicon, followed by microsatellite analysis and amplicon mutation-site minisequencing analysis. Conclusions The single-tube multiplex-PCR format of this highly polymorphic microsatellite marker panel simplifies identification and selection of informative markers for linkage-based PGD of HEMA. Informative markers can also be easily co-amplified with mutation-containing F8 amplicons for combined mutation detection and linkage analysis.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Single‐tube tetradecaplex panel of highly polymorphic microsatellite markers < 1 Mb from F8 for simplified preimplantation genetic diagnosis of hemophilia A

Zhao

Chen

Tan

et al. 2017

Journal of Thrombosis and Haemostasis

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“…Mutations c.5587-93C>T and c.5999-277G>A were identified by in-vitro assays using pcDNA 3.1/V5-His-TOPO vector [ 28 ]. To our knowledge, this was the first report using pSPL3 plasmids as the vectors in identifying splice mutations of F8 gene, and it provided another further verification method in genetic, prenatal diagnosis and preimplantation genetic diagnosis of hemophilia A [ 29 ]. The pSPL3 exon trapping vector contains two exons SD and SA, and a functional intron, with transcription beginning following the SV40 promoter and ending at the late poly (A) signal [ 30 ].…”

Section: Discussionmentioning

confidence: 99%

An intronic mutation c.6430-3C>G in the F8 gene causes splicing efficiency and premature termination in hemophilia A

Xia

Lin

et al. 2018

Blood Coagulation &Amp; Fibrinolysis

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Hemophilia A is a bleeding disorder caused by coagulation factor VIII protein deficiency or dysfunction, which is classified into severe, moderate, and mild according to factor clotting activity. An overwhelming majority of missense and nonsense mutations occur in exons of F8 gene, whereas mutations in introns can also be pathogenic. This study aimed to investigate the effect of an intronic mutation, c.6430-3C>G (IVS22-3C>G), on pre-mRNA splicing of the F8 gene. We applied DNA and cDNA sequencing in a Chinese boy with hemophilia A to search if any pathogenic mutation in the F8 gene. Functional analysis was performed to investigate the effect of an intronic mutation at the transcriptional level. Human Splicing Finder and PyMol were also used to predict its effect. We found the mutation c.6430-3C>G (IVS22-3C>G) in the F8 gene in the affected boy, with his mother being a carrier. cDNA from the mother and pSPL3 splicing assay showed that the mutation IVS22-3C>G results in a two-nucleotide AG inclusion at the 3′ end of intron 22 and leads to a truncated coagulation factor VIII protein, with partial loss of the C1 domain and complete loss of the C2 domain. The in-silico tool predicted that the mutation induces altered pre-mRNA splicing by using a cryptic acceptor site in intron 22. The IVS22-3C>G mutation was confirmed to affect pre-mRNA splicing and produce a truncated protein, which reduces the stability of binding between the F8 protein and von Willebrand factor carrier protein due to the loss of an interaction domain.

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“…15,16 If available, preimplantation genetic diagnosis (PGD) identifies unaffected embryos, yet has limitations. 17 Chorionic villus sampling and amniocentesis may be offered to interested carriers; these should be performed at knowledgeable centres due to a 1%-2% associated risk of miscarriage. 15,18 If diagnosis is not established before delivery, sex determination via ultrasound and cord blood testing at birth for male infants are recommended.…”

Section: Op Timal B Leed Pre Venti On: Defining the Op Timal Haemosmentioning

confidence: 99%

Optimizing bleed prevention throughout the lifespan: Womb to Tomb

Gupta

Shapiro

2018

Haemophilia

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The focus of care providers, patients and families is the ability to tailor care for persons with haemophilia (PWH) across the lifespan. Care requires knowledge of the bleeding disorder and age-related complications, risk of therapeutic interventions, and evaluation of individual characteristics that contribute to outcomes. The ultimate goal is to live a normal life without the burden of bleeding, for PWH and carriers. A wide range of therapeutic options is required to achieve personalized care. Over the last decade, substantial therapeutic advantages have been achieved in the treatment of haemophilia that include the development of a robust array of factor concentrates, novel haemostatic agents, and increased knowledge and awareness of disease associated outcomes and risk factors. Significant strides on the road to accessible gene therapy have been realized. This increased range of therapeutic modalities provides options for development and implementation of care plans for each patient at each stage of life that are more flexible compared to prior care regimens. Paradigms for management of haemophilia are changing. As a community, we must work together to use these resources wisely, to learn from outcomes with new therapies and diagnostic tools, to assure all patients can achieve improved care and outcomes regardless of disease state or country of origin.

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Preimplantation genetic diagnosis of hemophilia A

Cited by 13 publications

References 41 publications

Single‐tube tetradecaplex panel of highly polymorphic microsatellite markers < 1 Mb from F8 for simplified preimplantation genetic diagnosis of hemophilia A

Single‐tube tetradecaplex panel of highly polymorphic microsatellite markers < 1 Mb from F8 for simplified preimplantation genetic diagnosis of hemophilia A

An intronic mutation c.6430-3C>G in the F8 gene causes splicing efficiency and premature termination in hemophilia A

Optimizing bleed prevention throughout the lifespan: Womb to Tomb

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