A novel diagnostic screen for defects in the Fanconi anemia pathway.

Abstract

Fanconi anemia (FA) is an autosomal recessive chromosomal instability syndrome characterized by congenital abnormalities, progressive bone marrow failure, and cancer predisposition. Although patients with FA are candidates for bone marrow transplantation or gene therapy, their phenotypic heterogeneity can delay or obscure diagnosis. The current diagnostic test for FA consists of cytogenetic quantitation of chromosomal breakage in response to diepoxybutane (DEB) or mitomycin C (MMC). Recent studies have elucidated a biochemical pathway for Fanconi anemia that culminates in the monoubiquitination of the FANCD2 protein. In the current study, we develop a new rapid diagnostic and subtyping FA assay amenable for screening broad populations at risk of FA. Primary lymphocytes were assayed for FANCD2 monoubiquitination by immunoblot. The absence of the monoubiquitinated FANCD2 isoform correlated with the diagnosis of FA by DEB testing in 11 known patients with FA, 37 patients referred for possible FA, and 29 healthy control subjects. Monoubiquitination of FANCD2 was normal in other bone marrow failure syndromes and chromosomal breakage syndromes. A combination of retroviral gene transfer and FANCD2 immunoblotting provides a rapid subtyping assay for patients newly diagnosed with FA. These new FA screening assays would allow efficient testing of broad populations at risk.

• Authors: Akiko Shimamura, Rocio Montes de Oca, John L Svenson, Nicholas Haining, Lisa A Moreau, David G Nathan, Alan D D'Andrea
• Journal: Blood (Blood) Vol. 100 Issue 13 Pg. 4649-54 (Dec 15 2002) ISSN: 0006-4971 [Print] United States
• PMID: 12393398 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.)

Chemical References

• DNA, Complementary
• Epoxy Compounds
• FANCD2 protein, human
• Fanconi Anemia Complementation Group D2 Protein
• Nuclear Proteins
• Ubiquitin
• Mitomycin
• diepoxybutane

Topics

• Blotting, Western
• Bone Marrow Diseases (blood)
• Cell Line, Transformed
• Chromosome Breakage
• Chromosomes, Human (drug effects)
• DNA, Complementary (genetics)
• Epoxy Compounds (pharmacology)
• Fanconi Anemia (blood, classification, diagnosis, genetics)
• Fanconi Anemia Complementation Group D2 Protein
• Genetic Heterogeneity
• Genetic Testing
• Genetic Vectors (genetics)
• Humans
• Lymphocytes (chemistry)
• Microscopy, Fluorescence
• Mitomycin (pharmacology)
• Nuclear Proteins (blood, metabolism)
• Protein Processing, Post-Translational
• Retroviridae (genetics)
• Transfection
• Ubiquitin (metabolism)