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Molecular and Immunologic Techniques in a Genetically Engineered Mouse Model of Gastrointestinal Stromal Tumor
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BRAF mutations in KIT/PDGFRA positive gastrointestinal stromal tumours (GISTs): Is their frequency underestimated?

Karin Jašek1, Barbora Váňová1, Marián Grendár1

  • 1Comenius University in Bratislava, Jessenius Faculty of Medicine in Martin (JFM CU), Biomedical Center Martin JFM CU, 036 01 Martin, Slovakia.

Pathology, Research and Practice
|August 25, 2020
PubMed

Insights

BRAF V600E mutations are found in some gastrointestinal stromal tumors (GISTs), even those with KIT/PDGFRA mutations. Detecting these BRAF mutations is crucial for treatment decisions, as they impact imatinib response.

Area of Science:

  • Oncology
  • Molecular Biology
  • Genetics

Background:

  • BRAF V600E mutations represent a distinct molecular event in gastrointestinal stromal tumors (GISTs).
  • GISTs with BRAF mutations often exhibit resistance to imatinib therapy, posing a therapeutic challenge.
  • Phenotypic and morphologic similarities between BRAF-mutated and KIT/PDGFRA-mutated GISTs can obscure diagnosis.

Purpose of the Study:

  • To detect BRAF V600E mutations in KIT/PDGFRA-positive GISTs.
  • To verify BRAF mutation findings using orthogonal molecular techniques.
  • To assess the diagnostic utility of droplet digital PCR (ddPCR) for BRAF mutations in GIST.

Main Methods:

  • Sensitive droplet digital PCR (ddPCR) was employed to detect BRAF V600E mutations in 35 KIT/PDGFRA-positive GIST samples.
  • Method sensitivity was established through rigorous determination of false positive rate (FPR) and limit of detection (LoD).
  • ddPCR results were validated using allele-specific PCR, dideoxysequencing, and competitive allele-specific TaqMan PCR (castPCR).

Main Results:

  • The ddPCR method demonstrated high sensitivity, with an LoD of 3.4293 copies/μL (0.0162% sensitivity).
  • Eight KIT/PDGFRA-positive GIST patients with concomitant BRAF V600E mutations were identified.
  • Concomitant mutations included five cases with KIT and three cases with PDGFRA mutations.

Conclusions:

  • Droplet digital PCR is a highly sensitive method for detecting BRAF V600E mutations in GISTs.
  • The identification of concomitant BRAF mutations in KIT/PDGFRA-positive GISTs has significant therapeutic implications.
  • Accurate BRAF mutation detection is essential for optimizing treatment strategies in GIST patients.
Keywords:
BRAF inhibitorBRAF mutationBRAF/KIT/PDGFRA genesConcomitant mutationsDroplet digital PCRGastrointestinal stromal tumours

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