A cross‐linker‐sensitive myeloid leukemia cell line from a 2‐year‐old boy with severe Fanconi anemia and biallelic <i>FANCD1</i>/<i>BRCA2</i> mutations

Meyer, Stefan; Fergusson, William; Oostra, Anneke B.; Medhurst, Annette L.; Waisfisz, Quinten; Winter, Johan P. de; Chen, Fei; Carr, T; Clayton‐Smith, Jill; Clancy, Tara; Green, Mike; Barber, Lisa M.; Eden, O. B.; Will, Andrew; Joenje, Hans; Taylor, G. Malcolm

doi:10.1002/gcc.20153

Cited by 27 publications

(36 citation statements)

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“…12,18 Table 1 gives a review of the 30 patients from 24 families with biallelic BRCA2 mutations and whose phenotype had previously been described in details family. 12,[19][20][21][22][23][24][25][26][27][28] Among these patients, the females and males were equally affected and the mean age of FA diagnosis was 1.9 years (range, 0.1-5.2 years). Most of the patients were diagnosed as FA because of their typical physical features (68%).…”

Section: Discussionmentioning

confidence: 99%

Development of primary early-onset colorectal cancers due to biallelic mutations of the FANCD1/BRCA2 gene

et al. 2013

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Fanconi anaemia (FA) is characterized by progressive bone marrow failure, congenital anomalies, and predisposition to malignancy. In a minority of cases, FA results from biallelic FANCD1/BRCA2 mutations that are associated with early-onset leukaemia and solid tumours. Here, we describe the clinical and molecular features of a remarkable family presenting with multiple primary colorectal cancers (CRCs) without detectable mutations in genes involved in the Mendelian predisposition to CRCs. We unexpectedly identified, despite the absence of clinical cardinal features of FA, a biallelic mutation of the FANCD1/BRCA2 corresponding to a frameshift alteration (c.1845_1846delCT, p.Asn615Lysfs*6) and a missense mutation (c.7802A4G, p.Tyr2601Cys). The diagnosis of FA was confirmed by the chromosomal analysis of lymphocytes. Reverse transcriptase (RT)-PCR analysis revealed that the c.7802A4G BRCA2 variation was in fact a splicing mutation that creates an aberrant splicing donor site and results partly into an aberrant transcript encoding a truncated protein (p.Tyr2601Trpfs*46). The atypical FA phenotype observed within this family was probably explained by the residual amount of BRCA2 with the point mutation c.7802A4G in the patients harbouring the biallelic FANCD1/BRCA2 mutations. Although this report is based in a single family, it suggests that CRCs may be part of the tumour spectrum associated with FANCD1/BRCA2 biallelic mutations and that the presence of such mutations should be considered in families with CRCs, even in the absence of cardinal features of FA.

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Section: Discussionmentioning

confidence: 99%

Development of primary early-onset colorectal cancers due to biallelic mutations of the FANCD1/BRCA2 gene

et al. 2013

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“…We found this splice variant to be expressed in an EBV-transformed lymphoblastoid cell line (AVO35) established from a healthy IVS7 ϩ 2T Ͼ G carrier 21 ( Figure 4A lane 3). We then tested the expression level of this alternatively spliced transcript For personal use only.…”

Section: Expression Of ⌬Exons 4-7 Splice Variant In Human Cellsmentioning

confidence: 93%

“…Human EBV-lymphoblastoid cell lines (AVO35), leukemia cell line (SB1685CB), and nonimmortilized fibroblasts (AC389) were maintained as described previously. 21 All oligonucleotides were obtained from Invitrogen, and their sequences are listed in supplemental Table 3. Antibodies used are: c-myc tag (ab18185, Abcam), actin (Ab-5, NeoMarkers), Rad51 (H92, Santa Cruz Biotechnology, sc8349), and ␥-H2AX (clone JBW103, Upstate Biotechnology).…”

Section: Reagentsmentioning

confidence: 99%

“…by guest www.bloodjournal.org From in the normal fibroblasts (AC389) and the leukemia cells (SB1685CB) of this carrier's son who was affected by FA and developed acute myeloid leukemia (AML) at the age of 2 years. 21 The patient was a BRCA2 compound heterozygote, inheriting the IVS7 ϩ 2T Ͼ G mutation from the father and a 2-bp deletion in exon 11 (c.3827delGT) from the mother. Interestingly, although the same transcripts were present in the child's normal fibroblast and leukemia cells as those detected in the father's cells, there was a marked quantitative difference in the expression of 2 transcripts.…”

Section: Expression Of ⌬Exons 4-7 Splice Variant In Human Cellsmentioning

confidence: 99%

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A comprehensive functional characterization of BRCA2 variants associated with Fanconi anemia using mouse ES cell–based assay

Biswas

Das

Alter

et al. 2011

Blood

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Biallelic mutations in the human breast cancer susceptibility gene, BRCA2, are associated with Fanconi anemia, implying that some persons who inherit 2 deleterious variants of BRCA2 are able to survive even though it is well established that BRCA2 is indispensable for viability in mice. One such variant, IVS7 ؉ 2T > G, results in premature protein truncation because of skipping of exon 7. Surprisingly, the persons who are either IVS7 ؉ 2T > G homozygous or compound heterozygous are born alive but die of malignancy associated with Fanconi anemia. Using a mouse embryonic stem cellbased functional assay, we found that the IVS7 ؉ 2T > G allele produces an alternatively spliced transcript lacking exons 4-7, encoding an in-frame BRCA2 protein with an internal deletion of 105 amino acids (BRCA2 ⌬105 ). We demonstrate that BRCA2 ⌬105 is proficient in homologous recombination-mediated DNA repair as measured by different functional assays.

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“…Chromatin immunoprecipitation (ChIP) was carried out on 3 AML cell lines, SB1690CB (EVI1 overexpressing), 22 Three independent experiments were carried out and according to manufacturer's protocol. In each experiment, more than 1% of the input from the positive control RPL30 (Histone H3 binding target) was precipitated (mean 2%, SD 0.78%).…”

Section: Chipmentioning

confidence: 99%

Aberrant DNA hypermethylation signature in acute myeloid leukemia directed by EVI1

Lugthart

Figueroa

Bindels

et al. 2011

Blood

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DNA methylation patterns are frequently dysregulated in cancer, although little is known of the mechanisms through which specific gene sets become aberrantly methylated. The ecotropic viral integration site 1 (EVI1) locus encodes a DNA binding zinc-finger transcription factor that is aberrantly expressed in a subset of acute myeloid leukemia (AML) patients with poor outcome. We find that the promoter DNA methylation signature of EVI1 AML blast cells differs from those of normal CD34 ؉ bone marrow cells and other AMLs. This signature contained 294 differentially methylated genes, of which 238 (81%) were coordinately hypermethylated. An unbiased motif analysis revealed an overrepresentation of EVI1 binding sites among these aberrantly hypermethylated loci. EVI1 was capable of binding to these promoters in 2 different EVI1-expressing cell lines, whereas no binding was observed in an EVI1-negative cell line. Furthermore, EVI1 was observed to interact with DNA methyl transferases 3A and 3B. Among the EVI1 AML cases, 2 subgroups were recognized, of which 1 contained AMLs with many more methylated genes, which was associated with significantly higher levels of EVI1 than in the cases of the other subgroup. Our data point to a role for EVI1 in directing aberrant promoter DNA methylation patterning in EVI1 AMLs. (Blood. 2011;117(1):234-241) IntroductionPatterning of DNA methylation plays a critical role in epigenetic gene regulation during normal development. 1 Aberrant cytosine methylation of gene promoters occurs frequently in many forms of cancer, including acute myeloid leukemias (AMLs). 2 Several tumor suppressor genes (eg, CDKN2B and CEBPA) are found to be abnormally methylated and silenced in AML patients. 3,4 Moreover, aberrant distribution of promoter DNA methylation occurring in specific and distinct patterns has been shown to be a universal feature occurring in all AML patients. 5 However, the mechanisms that mediate these aberrant methyl cytosine patterns have not been defined.Abnormal expression of the EVI1 (ecotropic viral integration site 1) gene, as the result of inv(3)(q21q26.2)/t(3,3)(q21;q26.2) or through other unknown mechanisms, is associated with unfavorable AML outcome. 6,7 EVI1 encodes a C2H2 zinc finger transcription factor, binding DNA in a sequence-specific manner and functions as a repressor. 8-10 Retroviral insertion mutagenesis studies suggest that EVI1 deregulation plays a role in leukemogenesis. 11 The mechanism through which EVI1 mediates these effects is unknown.Given the function of EVI1 as a transcriptional repressor, we wondered whether EVI1 might be associated with aberrant epigenetic programming in AML patients. To test this hypothesis, we conducted a large-scale DNA methylation profiling study in human EVI1 AMLs. A specific promoter DNA methylation signature was uncovered in EVI1 AMLs, and evidence was provided that EVI1 contributes to aberrant promoter DNA methylation patterning in those leukemias. Methods Patient samplesDiagnostic material from 26 AML patients overexpressing EVI1 ...

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A cross‐linker‐sensitive myeloid leukemia cell line from a 2‐year‐old boy with severe Fanconi anemia and biallelic FANCD1/BRCA2 mutations

Cited by 27 publications

References 27 publications

Development of primary early-onset colorectal cancers due to biallelic mutations of the FANCD1/BRCA2 gene

Development of primary early-onset colorectal cancers due to biallelic mutations of the FANCD1/BRCA2 gene

A comprehensive functional characterization of BRCA2 variants associated with Fanconi anemia using mouse ES cell–based assay

Aberrant DNA hypermethylation signature in acute myeloid leukemia directed by EVI1

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