The objective of this proposal is to isolate and characterize the genes defective in Fanconi Anemia (FA). Patients with this hereditary disorder have a varied set of congenital malformations, bone marrow failure and an increased risk for the development of myeloid leukemia. Thus, the genes defective in FA will reveal information about developmental processes in addition to cancer biogenesis. We have shown that there are at least four complementation groups in FA (A to D) and have developed an episomal cDNA functional complementation method, that exploits FA hypersensitivity to DNA-crosslinking agents, to isolate the defective genes. Recently, we have successfully used this method to isolate the gene for FA(C) and found that it codes for a novel protein. It is thus important to establish its functional relationship to the genes in the other FA groups. Two main projects are proposed. First, we will use the same methodology to clone the genes defective in the A, B and D groups. Candidate cDNAs will be tested for their ability to specifically correct the cellular defect in the same FA cells. The assignment will be confirmed by finding mutations in FA patients of the same group. Second, we will characterize the FA genes and their protein products. We will map the cDNA in the human genome, isolate the gene and characterize the intron-exon structure. we will raise antibodies against the predicted protein and use these to establish its physical properties and cellular location. The cell, tissue and developmental expression of the gene will established by RNA and in situ hybridization analysis of human and rodent samples. This research will have major impact in our understanding of the basic defect in this complex disorder. In the first instance, it will be possible to classify the various patients according to complementation group and to establish correlations between the manifestation of their disease and the underlying genetic defects. Second, DNA based tests will be developed for accurate diagnosis as well as for carrier and prenatal testing. In the longer term, as knowledge about the genes and their expression is developed, it may be feasible to consider gene therapy as a method of treating FA patients.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL050131-01
Application #
3369148
Study Section
Mammalian Genetics Study Section (MGN)
Project Start
1993-05-01
Project End
1996-04-30
Budget Start
1993-05-01
Budget End
1994-04-30
Support Year
1
Fiscal Year
1993
Total Cost
Indirect Cost
Name
Hospital for Sick Chldrn (Toronto)
Department
Type
DUNS #
208511808
City
Toronto
State
ON
Country
Canada
Zip Code
M5 1-X8
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de Winter, J P; Leveille, F; van Berkel, C G et al. (2000) Isolation of a cDNA representing the Fanconi anemia complementation group E gene. Am J Hum Genet 67:1306-8
Lightfoot, J; Alon, N; Bosnoyan-Collins, L et al. (1999) Characterization of regions functional in the nuclear localization of the Fanconi anemia group A protein. Hum Mol Genet 8:1007-15
Carreau, M; Alon, N; Bosnoyan-Collins, L et al. (1999) Drug sensitivity spectra in Fanconi anemia lymphoblastoid cell lines of defined complementation groups. Mutat Res 435:103-9
de Winter, J P; Waisfisz, Q; Rooimans, M A et al. (1998) The Fanconi anaemia group G gene FANCG is identical with XRCC9. Nat Genet 20:281-3
Lo Ten Foe, J R; Rooimans, M A; Bosnoyan-Collins, L et al. (1996) Expression cloning of a cDNA for the major Fanconi anaemia gene, FAA. Nat Genet 14:320-3