B-RAF mutant alleles associated with Langerhans cell histiocytosis, a granulomatous pediatric disease.
Langerhans cell histiocytosis (LCH) features inflammatory granuloma characterised by the presence of CD1a+ dendritic cells or 'LCH cells'. Badalian-Very et al. recently reported the presence of a canonical (V600E)B-RAF mutation in 57% of paraffin-embedded biopsies from LCH granuloma. Here...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2012-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3323620?pdf=render |
| _version_ | 1818323723300634624 |
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| author | Takeshi Satoh Alexander Smith Aurelien Sarde Hui-chun Lu Syed Mian Celine Trouillet Ghulam Mufti Jean-Francois Emile Franca Fraternali Jean Donadieu Frederic Geissmann |
| author_facet | Takeshi Satoh Alexander Smith Aurelien Sarde Hui-chun Lu Syed Mian Celine Trouillet Ghulam Mufti Jean-Francois Emile Franca Fraternali Jean Donadieu Frederic Geissmann |
| author_sort | Takeshi Satoh |
| collection | DOAJ |
| description | Langerhans cell histiocytosis (LCH) features inflammatory granuloma characterised by the presence of CD1a+ dendritic cells or 'LCH cells'. Badalian-Very et al. recently reported the presence of a canonical (V600E)B-RAF mutation in 57% of paraffin-embedded biopsies from LCH granuloma. Here we confirm their findings and report the identification of two novel B-RAF mutations detected in LCH patients.Mutations of B-RAF were observed in granuloma samples from 11 out of 16 patients using 'next generation' pyrosequencing. In 9 cases the mutation identified was (V600E)B-RAF. In 2 cases novel polymorphisms were identified. A somatic (600DLAT)B-RAF insertion mimicked the structural and functional consequences of the (V600E)B-RAF mutant. It destabilized the inactive conformation of the B-RAF kinase and resulted in increased ERK activation in 293 T cells. The (600DLAT)B-RAF and (V600E)B-RAF mutations were found enriched in DNA and mRNA from the CD1a+ fraction of granuloma. They were absent from the blood and monocytes of 58 LCH patients, with a lower threshold of sequencing sensitivity of 1%-2% relative mutation abundance. A novel germ line (T599A)B-RAF mutant allele was detected in one patient, at a relative mutation abundance close to 50% in the LCH granuloma, blood monocytes and lymphocytes. However, (T599A)B-RAF did not destabilize the inactive conformation of the B-RAF kinase, and did not induce increased ERK phosphorylation or C-RAF transactivation.Our data confirmed presence of the (V600E)B-RAF mutation in LCH granuloma of some patients, and identify two novel B-RAF mutations. They indicate that (V600E)B-RAF and (600DLAT)B-RAF mutations are somatic mutants enriched in LCH CD1a(+) cells and absent from the patient blood. Further studies are needed to assess the functional consequences of the germ-line (T599A)B-RAF allele. |
| first_indexed | 2024-12-13T11:17:13Z |
| format | Article |
| id | doaj.art-1906c4b7fd534969a6c38a88f95943be |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-13T11:17:13Z |
| publishDate | 2012-01-01 |
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| spelling | doaj.art-1906c4b7fd534969a6c38a88f95943be2022-12-21T23:48:36ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0174e3389110.1371/journal.pone.0033891B-RAF mutant alleles associated with Langerhans cell histiocytosis, a granulomatous pediatric disease.Takeshi SatohAlexander SmithAurelien SardeHui-chun LuSyed MianCeline TrouilletGhulam MuftiJean-Francois EmileFranca FraternaliJean DonadieuFrederic GeissmannLangerhans cell histiocytosis (LCH) features inflammatory granuloma characterised by the presence of CD1a+ dendritic cells or 'LCH cells'. Badalian-Very et al. recently reported the presence of a canonical (V600E)B-RAF mutation in 57% of paraffin-embedded biopsies from LCH granuloma. Here we confirm their findings and report the identification of two novel B-RAF mutations detected in LCH patients.Mutations of B-RAF were observed in granuloma samples from 11 out of 16 patients using 'next generation' pyrosequencing. In 9 cases the mutation identified was (V600E)B-RAF. In 2 cases novel polymorphisms were identified. A somatic (600DLAT)B-RAF insertion mimicked the structural and functional consequences of the (V600E)B-RAF mutant. It destabilized the inactive conformation of the B-RAF kinase and resulted in increased ERK activation in 293 T cells. The (600DLAT)B-RAF and (V600E)B-RAF mutations were found enriched in DNA and mRNA from the CD1a+ fraction of granuloma. They were absent from the blood and monocytes of 58 LCH patients, with a lower threshold of sequencing sensitivity of 1%-2% relative mutation abundance. A novel germ line (T599A)B-RAF mutant allele was detected in one patient, at a relative mutation abundance close to 50% in the LCH granuloma, blood monocytes and lymphocytes. However, (T599A)B-RAF did not destabilize the inactive conformation of the B-RAF kinase, and did not induce increased ERK phosphorylation or C-RAF transactivation.Our data confirmed presence of the (V600E)B-RAF mutation in LCH granuloma of some patients, and identify two novel B-RAF mutations. They indicate that (V600E)B-RAF and (600DLAT)B-RAF mutations are somatic mutants enriched in LCH CD1a(+) cells and absent from the patient blood. Further studies are needed to assess the functional consequences of the germ-line (T599A)B-RAF allele.http://europepmc.org/articles/PMC3323620?pdf=render |
| spellingShingle | Takeshi Satoh Alexander Smith Aurelien Sarde Hui-chun Lu Syed Mian Celine Trouillet Ghulam Mufti Jean-Francois Emile Franca Fraternali Jean Donadieu Frederic Geissmann B-RAF mutant alleles associated with Langerhans cell histiocytosis, a granulomatous pediatric disease. PLoS ONE |
| title | B-RAF mutant alleles associated with Langerhans cell histiocytosis, a granulomatous pediatric disease. |
| title_full | B-RAF mutant alleles associated with Langerhans cell histiocytosis, a granulomatous pediatric disease. |
| title_fullStr | B-RAF mutant alleles associated with Langerhans cell histiocytosis, a granulomatous pediatric disease. |
| title_full_unstemmed | B-RAF mutant alleles associated with Langerhans cell histiocytosis, a granulomatous pediatric disease. |
| title_short | B-RAF mutant alleles associated with Langerhans cell histiocytosis, a granulomatous pediatric disease. |
| title_sort | b raf mutant alleles associated with langerhans cell histiocytosis a granulomatous pediatric disease |
| url | http://europepmc.org/articles/PMC3323620?pdf=render |
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