Identification of two complementation groups in Fanconi anemia

1985 ◽  
Vol 11 (1) ◽  
pp. 35-41 ◽  
Author(s):  
G. Duckworth-Rysiecki ◽  
K. Cornish ◽  
C. A. Clarke ◽  
M. Buchwald
Keyword(s):  
Fanconi Anemia ◽  
Complementation Groups

The IVS4 + 4 A to T mutation of the Fanconi anemia geneFANCC is not associated with a severe phenotype in Japanese patients

Blood ◽  
2000 ◽  
Vol 95 (4) ◽  
pp. 1493-1498 ◽  
Author(s):  
Makoto Futaki ◽  
Takayuki Yamashita ◽  
Hiroshi Yagasaki ◽  
Tatsushi Toda ◽  
Miharu Yabe ◽  
...  
Keyword(s):  
Fanconi Anemia ◽  
Clinical Phenotype ◽  
Japanese Patients ◽  
Single Strand Conformation Polymorphism ◽  
Common Mutation ◽  
Ashkenazi Jewish ◽  
Severe Phenotype ◽  
Significant Difference ◽  
Complementation Groups ◽  
Intron 4

Fanconi anemia (FA) is an autosomal recessive disease characterized by congenital anomalies, aplastic anemia, and a susceptibility to leukemia. There are at least 8 complementation groups (A through H). Extensive analyses of the FA group C gene FANCC in Western countries revealed that 10% to 15% of FA patients have mutations of this gene. The most common mutation is IVS4 + 4 A to T (IVS4), a splice mutation in intron 4, which has been found only in patients of Ashkenazi Jewish ancestry. When we screened 29 Japanese patients (20 unrelated patients and 4 families) using polymerase chain reaction–single strand conformation polymorphism, we found 8 unrelated patients homozygous for IVS4. This is apparently the first non–Ashkenazi-Jewish population for whom this mutation has been detected. The Ashkenazi Jewish patients homozygous for IVS4 have a severe phenotype, in comparison with other FA patients. Our analyses of Japanese patients indicate no significant difference between IVS4 homozygotes and other patients with regard to severity of a clinical phenotype. Thus, ethnic background may have a significant effect on a clinical phenotype in FA patients carrying the same mutation.

Protein Replacement by Receptor-Mediated Endocytosis Corrects the Sensitivity of Fanconi Anemia Group C Cells to Mitomycin C

Blood ◽  
1999 ◽  
Vol 93 (1) ◽  
pp. 363-369 ◽  
Author(s):  
Hagop Youssoufian ◽  
Frank A.E. Kruyt ◽  
Xiaotong Li
Keyword(s):  
Mitomycin C ◽  
Fanconi Anemia ◽  
Hematopoietic Cells ◽  
Chimeric Protein ◽  
Direct Gene Transfer ◽  
C Cells ◽  
Receptor Mediated Endocytosis ◽  
Culture Model ◽  
Complementation Groups ◽  
Anemia Group

Abstract Current methods for direct gene transfer into hematopoietic cells are inefficient. Here we show that functional complementation of Fanconi anemia (FA) group C cells by protein replacement can be as efficacious as by transfection with wild-type FAC cDNA. We expressed a chimeric protein (called His-ILFAC) consisting of the mature coding portion of gibbon interleukin-3 (IL-3) and full-length FAC inEscherichia coli. The purified bacterial protein is internalized by hematopoietic cells via IL-3 receptors. The intracellular half-life of His-ILFAC is approximately 60 minutes, which is comparable to that of the transgene-encoded FAC protein. In this cell-culture model His-ILFAC completely corrects the sensitivity of FA group C cells to mitomycin C, but it has no effect on FA cells that belong to complementation groups A and B. We suggest that receptor-mediated endocytosis of cytokine-fusion proteins may be of general use to deliver macromolecules into hematopoietic progenitor cells.

scholarly journals Characterization of Pathogenic Variants and Clinical Phenotypes in 117 Japanese Fanconi Anemia Patients

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 3860-3860
Author(s):  
Minako Mori ◽  
Asuka Hira ◽  
Kenichi Yoshida ◽  
Hideki Muramatsu ◽  
Yusuke Okuno ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Exome Sequencing ◽  
Molecular Diagnosis ◽  
Fanconi Anemia ◽  
Direct Sequencing ◽  
Bone Marrow Failure ◽  
Whole Genome ◽  
Aberrant Splicing ◽  
Large Deletions ◽  
Complementation Groups

Abstract Objective: Fanconi anemia (FA) is the most common inherited bone marrow failure syndrome associated with multiple congenital abnormalities and predisposition to malignancies, resulting from mutations in one of the 22 known FA genes (FANCA to W). The proteins encoded by these genes participate in DNA repair pathway (the FA pathway) for endogenous aldehyde damage. Compared to the situation in the US or Europe, the number of Japanese FA patients with genetic diagnosis was relatively limited. In this study, we reveal the genetic subtyping and the characteristics of mutated FA genes in Japanese population and clarify the genotype-phenotype correlations. Results: We studied 117 Japanese FA patients from 103 families (1996 to 2018). The diagnosis of FA was confirmed on the basis of chromosomal breakage tests and clinical features. Molecular diagnosis was obtained in 107 (91.5%) of the 117 patients through direct sequencing of FANCA and FANCG, MLPA analysis for FANCA, targeted exome sequencing (targeted-seq), and whole exome sequencing (WES) analysis (Figure 1). To provide genetic subtyping for the 10 unclassified cases, we tried to apply various technologies. Array CGH revealed large deletions in two FA-B and one FA-T cases. Whole genome sequencing and RNA-sequencing analysis identified splicing site or aberrant splicing mutations among three cases (one FA-B, one FA-C, and one FA-N). Collectively, 113 (97%) of Japanese 117 FA patients were successfully subtyped and a total of 219 mutated alleles were identified. FA-A and FA-G accounted for the disease in 58% and 25% of FA patients, respectively, whereas each of the other complementation groups accounted for less than 5% of FA cases. FANCB was the third most common complementation group (n=4) and only one FA-C case was identified in Japanese FA patients. In the 68 FA-A patients, we identified 130 mutant alleles that included 55 different FANCA variants (17 nucleotide substitutions, 16 small deletions/insertions, 12 large deletions, 1 large duplication and 9 splice site mutation). FANCA c.2546delC was the most prevalent (41/130 alleles; 32%). In the 29 FA-G patients, 57 mutant alleles were identified and seven different FANCG variants were detected. FANCG c.307+1G>C and 1066C>T accounted for most of FANCG mutant alleles (49/57; 88%) in the Japanese FA-G patients. The three hotspot mutations (FANCA c.2546delC, FANCG c.307+1G>C and c.1066C>T) existed at low prevalence (0.04-0.1%) in the whole-genome reference panel of 3554 Japanese individuals (3.5KJPN, Tohoku Megabank). Consistent with the paucity of the FA-C patients as opposed to the previous report (Blood 2000), the FANCC IVS4+4A mutation was absent in the 3.5KJPN database. We were able to examine the hematological outcomes in a subset of our cases (52 FA-A and 23 FA-G). Interestingly, the FA-G patients developed bone marrow failure (BMF) at a significantly younger age than FA-A patients (median age at onset of BMF: 3.1 years vs 5 years). Furthermore, the patients with the FANCA c.2546delC mutation had an increased risk of developing myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML), compared to FA-A patients without the mutation. In the rare complementation groups of FA, two FA-B cases with complete loss of FANCB gene and one FA-I patient with N-terminal premature termination codons revealed severe somatic abnormalities, consistent with VACTERL-H association. Two FANCD1 (BRCA2) patients and one FANCN (PALB2) patients did not experience bone marrow failure but developed early-onset malignancies (immature teratoma, T-lymphoblastic lymphoma, adenosquamous lung carcinoma, Wilms tumor). Conclusion: This is the largest series of subtyped Japanese FA patients to date and the results would be useful for future clinical management. To provide molecular diagnosis for FA in Japan, we suggest to start with PCR-direct sequencing of the three common mutations (FANCA c.2546delC, FANCG c.307+1G>C and FANCG c.1066C>T) along with MLPA assay for FANCA. These analyses would enable the identification of about 50% of the mutant alleles. For the rest of the cases, WES or targeted-seq analysis should be useful, however, large deletions and aberrant splicing need to be kept in mind. Disclosures Takaori-Kondo: Pfizer: Honoraria; Novartis: Honoraria; Celgene: Honoraria, Research Funding; Bristol-Myers Squibb: Honoraria; Janssen Pharmaceuticals: Honoraria.

scholarly journals Functional Activity of the Fanconi Anemia Protein FAA Requires FAC Binding and Nuclear Localization

1998 ◽  
Vol 18 (10) ◽  
pp. 5952-5960 ◽  
Author(s):  
Dieter Näf ◽  
Gary M. Kupfer ◽  
Ahmed Suliman ◽  
Kathleen Lambert ◽  
Alan D. D’Andrea
Keyword(s):  
Nuclear Localization ◽  
Fanconi Anemia ◽  
Functional Activity ◽  
Simian Virus 40 ◽  
T Antigen ◽  
Nuclear Accumulation ◽  
Wild Type ◽  
Amino Terminal ◽  
Complementation Groups ◽  
Mutant Forms

ABSTRACT Fanconi anemia (FA) is an autosomal recessive disease characterized by genomic instability, cancer susceptibility, and cellular hypersensitivity to DNA-cross-linking agents. Eight complementation groups of FA (FA-A through FA-H) have been identified. Two FA genes, corresponding to complementation groups FA-A and FA-C, have been cloned, but the functions of the encoded FAA and FAC proteins remain unknown. We have recently demonstrated that FAA and FAC interact to form a nuclear complex. In this study, we have analyzed a series of mutant forms of the FAA protein with respect to functional activity, FAC binding, and nuclear localization. Mutation or deletion of the amino-terminal nuclear localization signal (NLS) of FAA results in loss of functional activity, loss of FAC binding, and cytoplasmic retention of FAA. Replacement of the NLS sequence with a heterologous NLS sequence, derived from the simian virus 40 T antigen, results in nuclear localization but does not rescue functional activity or FAC binding. Nuclear localization of the FAA protein is therefore necessary but not sufficient for FAA function. Mutant forms of FAA which fail to bind to FAC also fail to promote the nuclear accumulation of FAC. In addition, wild-type FAC promotes the accumulation of wild-type FAA in the nucleus. Our results suggest that FAA and FAC perform a concerted function in the cell nucleus, required for the maintenance of chromosomal stability.

scholarly journals Fanconi Anemia Proteins FANCA, FANCC, and FANCG/XRCC9 Interact in a Functional Nuclear Complex

1999 ◽  
Vol 19 (7) ◽  
pp. 4866-4873 ◽  
Author(s):  
Irene Garcia-Higuera ◽  
Yanan Kuang ◽  
Dieter Näf ◽  
Jennifer Wasik ◽  
Alan D. D’Andrea
Keyword(s):  
Nuclear Localization ◽  
Fanconi Anemia ◽  
Autosomal Recessive ◽  
Nuclear Protein ◽  
Cancer Susceptibility ◽  
Clinical Phenotype ◽  
Normal Cells ◽  
Amino Terminal ◽  
Nuclear Complex ◽  
Complementation Groups

ABSTRACT Fanconi anemia (FA) is an autosomal recessive cancer susceptibility syndrome with at least eight complementation groups (A to H). Three FA genes, corresponding to complementation groups A, C, and G, have been cloned, but their cellular function remains unknown. We have previously demonstrated that the FANCA and FANCC proteins interact and form a nuclear complex in normal cells, suggesting that the proteins cooperate in a nuclear function. In this report, we demonstrate that the recently cloned FANCG/XRCC9 protein is required for binding of the FANCA and FANCC proteins. Moreover, the FANCG protein is a component of a nuclear protein complex containing FANCA and FANCC. The amino-terminal region of the FANCA protein is required for FANCG binding, FANCC binding, nuclear localization, and functional activity of the complex. Our results demonstrate that the three cloned FA proteins cooperate in a large multisubunit complex. Disruption of this complex results in the specific cellular and clinical phenotype common to most FA complementation groups.

Comprehensive molecular study identified 12 complementation groups with 56 novel FANC gene variants in Indian Fanconi anemia subjects

2021 ◽  
Author(s):  
Merin George ◽  
Avani Solanki ◽  
Niranjan Chavan ◽  
Aruna Rajendran ◽  
Revathi Raj ◽  
...  
Keyword(s):  
Fanconi Anemia ◽  
Molecular Study ◽  
Gene Variants ◽  
Complementation Groups

The Cellular Control Enzyme PolyADP Ribosyl Transferase Is Eliminated in Cultured Fanconi Anemia Fibroblasts at Confluency

2003 ◽  
Vol 384 (1) ◽  
pp. 169-174 ◽  
Author(s):  
M.H. Ramirez ◽  
C. Adelfalk ◽  
M. Kontou ◽  
M. Hirsch-Kauffmann ◽  
M. Schweiger
Keyword(s):  
Oxidative Stress ◽  
Cell Death ◽  
Redox Potential ◽  
Cell Survival ◽  
Fanconi Anemia ◽  
Hereditary Disease ◽  
Pathogenic Mechanisms ◽  
Complementation Groups ◽  
Cellular Control

AbstractFanconi anemia (FA) is a hereditary disease of unknown pathogenic mechanisms, although mutations in seven different genes can be causative. Six of these genes have been cloned and sequenced. Only slight homology to the DNA of any other known gene has been found with the exception of FANCG which is identical to XRCC9. The function of these genes, including XRCC9, is presently unknown. Since pADP ribosyl transferase (pADPRT) plays a role in apoptosis, and apoptosis is affected in FA cells, we studied the correlation between pADPRT and FA cells. We reinvestigated the previously reported lack of pADPRT activity in fibroblasts from patients with Fanconi anemia. Here we describe the role of the lower redox potential of FA cells and demonstrate that this is an efficient strategy in the prevention of cell death due to the lack of energy under oxidative stress. This strategy is advantageous for the cells under the nonreplicative condition of confluency in which the risk of mutation is low and the prevention of apoptosis permits cell survival. pADPRT is not diminished to the same extent in all complementation groups of FA. It is prominent in FANCA, FANCG and FANCF cells, indicating that these genes control pADPRT diminution. Our experiments suggest that the pADPRT level is linked with the oxidoreduction reactions seen in FA.

A Rapid Method for Retroviral Mediated Subtyping of Complementation Group in Fanconi Anemia Patients.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 5261-5261
Author(s):  
Saurabh Chandra ◽  
Orna Levran ◽  
Ingrid Jurickova ◽  
Chiel Mass ◽  
Rick Kapur ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Lines ◽  
Fanconi Anemia ◽  
Mutational Analysis ◽  
High Titer ◽  
Retroviral Vectors ◽  
Complementation Group ◽  
Cell Cycle Analysis ◽  
G Cells ◽  
Complementation Groups

Abstract Fanconi anemia (FA) is a rare autosomal recessive disorder that results from mutations in at least eleven different genes (A, B, C, D1, D2, E, F, G, I, J, L). Recent studies have demonstrated that clinical progression of the disease may be influenced by inter- and intra-genic variations, underlining the importance of subtyping complementation groups. Previously we have utilized retroviral mediated gene transfer of FA cDNAs and subsequent analysis for the correction of hypersensitivity to Mitomycin C, to develop a valid method for subtyping of groups (Hanenberg et al., 2002). However, for lymphoblastic cell lines (LCL), G418 selection was required to compensate for low transduction efficiencies achieved. In the present study we have utilized RD114/GALV psuedotyped (high titer stable producer clones) bicistronic retroviral vectors that coexpress EGFP. This results in higher transduction efficiencies in LCLs (20–70% for C, F, G and 10–20 % for A vector) and allows for specific analysis of transduced EGFP-positive cells within the bulk cultures, thus obviating the need for selection. In addition, the assay relies on the correction of G2/M arrest that is analyzed by FLOW, further reducing the assay time. After transduction with cDNA-containing vectors for groups A, C, F and G, cells are exposed for 48 hours to the alkylating agent Melphalan, stained with Hoechst 33342 dye and cell cycle analysis for FA-specific G2/M arrest is performed by flow cytometry. The percentage of EGFP-positive cells in the G2/M phase of cell cycle for each vector is calculated and compared using the MODFIT software. A complementation group is identified when correction in the G2/M arrest occurs. Results obtained with the assay matched the complementation group known for 12 control LCLs tested. We report here the results obtained for 39 FA patients with unknown complementation groups using this new assay. Of the 39 cell lines tested, complementation group was identified for 20 lines [FA-A(17), FA-C(1), FA-F(1), FA-G(1)], while 15 lines were typed as Non A/C/F/G. Cells from 4 patients did not undergo G2/M arrest. Mutation analysis by direct sequencing of genomic DNA after DHPLC completed for 11 patients thus far, has detected a mutation in one or both alleles confirming the results of the rapid complementation assay in every case (Table). For the 15 cell lines typed as Non A/C/F/G, additional studies including vectors for the FA genes FANCD2, FANCE, FANCL are underway. In summary, we have developed a rapid and efficient method for the identification of complementation groups in FA patients that also serves as a guide for quick mutational analysis. This methodology can also be applied to myeloid progenitor cells as well as fibroblasts, an advantage in patients with lymphoid mosaicism. Table 1. Mutational analysis of cell lines identified by cell cycle analysis Cell Line Ref # Cell cycle analysis Mutation 1(paternal) Mutation 2 (maternal) RA2593 A c.1471-?_2151+?del c.1471-?_2151+?del RA2483 A c.1901-?_2014+?del RA2625 A c.3239G>T(p.R1080L) RA2644 A c.2806G>A(p.E936K) c.2806G>A(p.E936K) RA2685 A c.2678G>A(p.W893X) RA2698 A c.337_338delTC RA2704 A c.3514-?_3626+?del c.3520_3522delTGG RA2724 C c.322delG c.711+4A>T(IVS4+4A>T) RA2191 A c.3520_3522dellTGG RA2709 F c.484-485delCT c.219delG RA2727 G c.908T>C c.908T>C

Israeli Fanconi Anemia Registry

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4125-4125
Author(s):  
Hannah Tamary ◽  
Blanche P. Alter ◽  
Daniella Nishri ◽  
Philip S Rosenberg
Keyword(s):  
Bone Marrow ◽  
Solid Tumors ◽  
Fanconi Anemia ◽  
Congenital Abnormality ◽  
Bone Marrow Failure ◽  
Epidemiological Data ◽  
Quantitative Model ◽  
Complementation Group ◽  
Adverse Outcomes ◽  
Complementation Groups

Abstract Using epidemiological data from retrospective cohorts of patients with Fanconi Anemia (FA) in North America and Germany a quantitative model to estimate bone marrow failure (BMF) and cancer risk was previously generated. To evaluate generalizability to another population, and to determine the risks for adverse outcomes in Israel, we created an Israeli FA registry and used the model to evaluate complications. We reviewed patient charts of 66 patients with FA diagnosed in Israel between 1964–2005. The data base included demographic information, as well as data describing the congenital abnormalities, FA complementation groups, BMT course and malignancies. Thirty six (36) patients were of Jewish origin [Ashkenzi 7, Sephardic 23, mixed 6] and 30 of Arabic origin. The first adverse event was bone marrow failure (BMF) in 35 patients (53%), hematological malignancy in 7 (11%) and 2 solid tumors in each of 3 patients (5%). The cause-specific hazard of BMF peaked at 10.5%/year at age 10 years (95% CI: 6.7–14.1%/year). The hazard of AML/ALL and MDS were stable at 0.9%/year (95% CI: 0.42–1.85%/year) and 1.4%/year (95% CI: 0.76–2.49%/year) respectively. The cumulative incidence of each outcome to age 32 was 70% for BMF, 13% for AML/ALL, and 17% for solid tumor. A five item congenital abnormality score was significantly associated with the risk of BMF (P = 0.009). The ratio of observed to expected cancer was 71 for all cancers [50 for solid tumors, 175 for leukemia] and >11,000 for myelodysplastic syndrome. Significantly elevated ratios of observed to expected cancers were observed for head and neck squamous cell carcinoma in 2 patients (986-fold), tumor of larynx (13,238-fold), vulva (3,701-fold), cervix (244-fold) and breast (88-fold). The complementation group was known in 41 patients [A 25 (63%), C 9 (22%), G 6 (15%), and D1 1 (2%)]. However, associations between complementation groups and specific outcomes were not significant. Despite the different ethnic background and the smaller number of FA patients in the Israeli cohort the risk estimates compared with the US and German cohorts were similar. As previously suggested the congenital abnormality score was significantly associated with the risk of BMF; an extraordinary risk of developing AML/MDS and later specific solid tumors was also found.

Protein Replacement by Receptor-Mediated Endocytosis Corrects the Sensitivity of Fanconi Anemia Group C Cells to Mitomycin C

Blood ◽  
1999 ◽  
Vol 93 (1) ◽  
pp. 363-369
Author(s):  
Hagop Youssoufian ◽  
Frank A.E. Kruyt ◽  
Xiaotong Li
Keyword(s):  
Mitomycin C ◽  
Fanconi Anemia ◽  
Hematopoietic Cells ◽  
Chimeric Protein ◽  
Direct Gene Transfer ◽  
C Cells ◽  
Receptor Mediated Endocytosis ◽  
Culture Model ◽  
Complementation Groups ◽  
Anemia Group

Current methods for direct gene transfer into hematopoietic cells are inefficient. Here we show that functional complementation of Fanconi anemia (FA) group C cells by protein replacement can be as efficacious as by transfection with wild-type FAC cDNA. We expressed a chimeric protein (called His-ILFAC) consisting of the mature coding portion of gibbon interleukin-3 (IL-3) and full-length FAC inEscherichia coli. The purified bacterial protein is internalized by hematopoietic cells via IL-3 receptors. The intracellular half-life of His-ILFAC is approximately 60 minutes, which is comparable to that of the transgene-encoded FAC protein. In this cell-culture model His-ILFAC completely corrects the sensitivity of FA group C cells to mitomycin C, but it has no effect on FA cells that belong to complementation groups A and B. We suggest that receptor-mediated endocytosis of cytokine-fusion proteins may be of general use to deliver macromolecules into hematopoietic progenitor cells.

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