Background: Pulmonary Langerhans cell histiocytosis is a localized proliferation of Langerhans cells in the lung that presents without systemic manifestations as bilateral nodular lung disease in adult cigarette smokers. The molecular basis for this proliferation is unknown.
Methods: Twenty-two concurrent nodules in five patients were microdissected from formalin-fixed paraffin-embedded tissue and analyzed by next-generation sequencing for mutations in 46 cancer genes with the Ion AmpliSeq Cancer Panel on an Ion PGM (Personal Genome Machine) Sequencer (Life Technologies Corporation). Mutation confirmation was performed by conventional Sanger sequencing or by sensitive coamplification at lower denaturation polymerase chain reaction/fluorescence melting curve analysis.
Results: Small amounts of DNA (10 ng) isolated from nodules were sufficient for successful interrogation of 740 mutational hot spots in 46 cancer genes by the Ion PGM Sequencer, with an average depth of coverage of 2,783 reads per hot spot and with uniformity of coverage of 92%. BRAF V600E mutation was detected in all concurrent nodules studied in two of the five patients, whereas in three of the five patients, no oncogene mutations were found.
Conclusions: Pulmonary Langerhans cell histiocytosis appears to be a clonal proliferation that may or may not have BRAF V600E mutations. For those with BRAF V600E mutations, new targeted therapies, such as vemurafenib, may be used in progressive cases.