Seven novel and four recurrent point mutations in the factor VIII (F8C) gene

10560 Background: Activating insertion mutations in exon 20 of EGFR are reported in a small subset of lung adenocarcinomas (ADC). In contrast to the classic EGFR mutations, they appear to confer primary resistance to currently approved EGFR tyrosine kinase inhibitors. Their incidence and clinicopathologic features are not well established. Methods: Lung ADCs (n=1500) were screened for major activating mutations in EGFR (exons 19 and 21) and KRAS (exon 2). Negative cases were tested for EGFR exon 20 insertions by a PCR-based sizing assay. Extended testing for additional recurrent point mutations in EGFR, KRAS, BRAF, NRAS, PIK3CA, MEK1 and AKT was performed in all cases by Sequenom mass spectrometry. A subset of cases was also tested for ALK rearrangements by FISH. Results: We identified 32cases withEGFRexon 20 insertions, accounting for 11% of all EGFR mutations. EGFRexon 20 insertions were mutually exclusive with the other genetic alterations tested except for PIK3CA mutations. The incidence was higher among never-smokers (p<0.0001) but there was no association with sex, ethnic origin or stage at diagnosis. Insertions were 3, 6, 9 or 12bp; 9bp insertions were most common (50%, 16/32). Morphologically, 90% of tumors were moderate to poorly differentiated with a predominant mixed ADC phenotype. Conclusions: EGFR exon 20 testing may identify a unique subset of EGFR mutant lung ADCs which is significantly larger than previously reported, making this the third most common type of EGFR mutation after exon 19 deletions and L858R. This population could potentially benefit from alternate targeted therapies, many of which are currently in clinical development.

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Use of denaturing gradient gel electrophoresis to detect point mutations in the factor VIII gene

Genomics ◽

10.1016/0888-7543(90)90569-g ◽

1990 ◽

Vol 6 (2) ◽

pp. 293-301 ◽

Cited By ~ 45

Author(s):

Monica D. Traytman ◽

Miyoko Higuchi ◽

Carol K. Kasper ◽

Stylianos E. Antonarakis ◽

Haig H. Kazazian

Keyword(s):

Factor Viii ◽

Gel Electrophoresis ◽

Denaturing Gradient Gel Electrophoresis ◽

Point Mutations ◽

Factor Viii Gene ◽

Gradient Gel Electrophoresis

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Eleven novel mutations in the factor VIII gene from Brazilian hemophilia A patients

Blood ◽

10.1182/blood.v86.8.3015.bloodjournal8683015 ◽

1995 ◽

Vol 86 (8) ◽

pp. 3015-3020

Author(s):

VR Arruda ◽

WC Pieneman ◽

PH Reitsma ◽

PP Deutz-Terlouw ◽

JM Annichino-Bizzacchi ◽

...

Keyword(s):

Factor Viii ◽

Southern Blotting ◽

Hemophilia A ◽

Severe Disease ◽

Point Mutations ◽

Electrophoretic Pattern ◽

Telomeric Region ◽

Factor Viii Gene ◽

Single Strand Conformational Polymorphism ◽

Almost All

The molecular characterization of the mutations in hemophilia A patients is hampered by the large size of the factor VIII gene and the great heterogeneity of mutations. In this study, we have performed a protocol involving multiplex polymerase chain reaction in which 19 exons were amplified in four different combinations followed by nonradioactive single-strand conformational polymorphism (SSCP) to screen for mutations. Southern blotting was used to detect inversion of the factor VIII gene resulting from recombination between copies of the gene A (F8A) located in intron 22 of the factor VIII gene and two copies close telomeric region of X chromosome. Forty-two hemophilia A patients (21 with severe and 21 with mild-to-moderate disease) were studied. The inversion of factor VIII occurred in 13 of 21 patients affected by severe hemophilia A. One patient showed a large extra band in addition to the three bands observed after Southern blotting with the F8A probe. An abnormal electrophoretic pattern of SSCP was detected in 85% and 50% of the patients affected by mild-to-moderate and severe disease, respectively. Sixteen different mutations were identified. Eleven mutations were novel and comprised 9 point mutations and 2 small deletions. This study shows that the methodology used is safe and rapid and has potential for detecting almost all of the genetic defects of the studied hemophilia A patients.

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A CATH domain functional family based approach to identify putative cancer driver genes and driver mutations

10.1101/399014 ◽

2018 ◽

Author(s):

Paul Ashford ◽

Camilla S.M. Pang ◽

Aurelio A. Moya-García ◽

Tolulope Adeyelu ◽

Christine A. Orengo

Keyword(s):

Zinc Finger Protein ◽

Point Mutations ◽

Driver Mutations ◽

Cancer Genes ◽

Driver Genes ◽

Recurrent Point ◽

Cancer Driver ◽

Functional Sites ◽

Cancer Driver Genes ◽

Family Based

Tumour sequencing identifies highly recurrent point mutations in cancer driver genes, but rare functional mutations are hard to distinguish from large numbers of passengers. We developed a novel computational platform applying a multi-modal approach to filter out passengers and more robustly identify putative driver genes. The primary filter identifies enrichment of cancer mutations in CATH functional families (CATH-FunFams) – structurally and functionally coherent sets of evolutionary related domains. Using structural representatives from CATH-FunFams, we subsequently seek enrichment of mutations in 3D and show that these mutation clusters have a very significant tendency to lie close to known functional sites or conserved sites predicted using CATH-FunFams. Our third filter identifies enrichment of putative driver genes in functionally coherent protein network modules confirmed by literature analysis to be cancer associated.Our approach is complementary to other domain enrichment approaches exploiting Pfam families, but benefits from more functionally coherent groupings of domains. Using a set of mutations from 22 cancers we detect 151 putative cancer drivers, of which 79 are not listed in cancer resources and include recently validated cancer genes EPHA7, DCC netrin-1 receptor and zinc-finger protein ZNF479.

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