DNA topoisomerases and the DNA lesion in human genetic instability syndromes

Genetically determined chromosomal instability entails, among other sequelae, a condition of elevated cancer risk. Patients with the autosomal recessive disorder Fanconi's anemia (FA) often develop leukemias of the monocytic lineage together with pancytopenia, whereas the Bloom's syndrome (BS) mutation confers an early and elevated incidence of neoplasia of no particular type. Cultured cells from FA patients show spontaneously elevated rates of chromosome aberrations and a hypersensitivity to DNA cross-linking agents. Cytogenetic evaluation of cells from BS patients revealed elevated rates of sister chromatid exchanges, which were sensitive to the bromodeoxyuridine (BrdU) concentration used in the assay. Such a BrdU sensitivity was also found in cultured cells from healthy subjects exposed to the intracellular superoxide generator paraquat or to bleomycin. Skin fibroblasts from FA and BS patients show poor growth, which in the case of FA could be mitigated by lowering the oxygen concentration to 5%. Lymphoblastoid B-cell lines derived from peripheral blood samples from FA and BS patients show elevated numbers of cells arrested in the G2 phase of the cell cycle. This phenomenon could also be provoked by exposing cell lines from healthy subjects to compounds interfering with the function of DNA topoisomerase I (camptothecin) or II (m-AMSA). To test for a putative deficiency of either DNA topoisomerase, B-cell cultures from FA and BS patients were compared with cell cultures from healthy subjects regarding their sensitivity towards camptothecin and m-AMSA. No difference in sensitivity to these agents was found in patient vs. control cell lines, thus ruling out a deficiency in DNA topoisomerase I or II as the prime defect in these conditions of elevated cancer risk. The similarity between the cell cycle kinetic patterns found in untreated FA cell lines and in normal cell lines exposed to camptothecin or m-AMSA suggest that the DNA lesion in FA, presumably being caused by an oxygen-related mechanism, may interfere with DNA topoisomerase function.