ABSTRACT
Androgens have been used in the treatment of bone marrow failure syndromes without a clear understanding of their mechanism of action. Blood counts of patients with dyskeratosis congenita or aplastic anemia with mutations in telomerase genes can improve with androgen therapy. Here we observed that exposure in vitro of normal peripheral blood lymphocytes and human bone marrow-derived CD34(+) cells to androgens increased telomerase activity, coincident with higher TERT mRNA levels. Cells from patients who were heterozygous for telomerase mutations had low baseline telomerase activity, which was restored to normal levels by exposure to androgens. Estradiol had an effect similar to androgens on TERT gene expression and telomerase enzymatic activity. Tamoxifen abolished the effects of both estradiol and androgens on telomerase function, and letrozole, an aromatase inhibitor, blocked androgen effects on telomerase activity. Conversely, flutamide, an androgen receptor antagonist, did not affect androgen stimulation of telomerase. Down-regulation by siRNA of estrogen receptor-alpha (ER alpha), but not ER beta, inhibited estrogen-stimulated telomerase function. Our results provide a mechanism for androgen therapy in bone marrow failure: androgens appear to regulate telomerase expression and activity mainly by aromatization and through ER alpha. These findings have potential implications for the choice of current androgenic compounds and the development of future agents for clinical use.
Subject(s)
Androgens/pharmacology , Estradiol/pharmacology , Estrogens/pharmacology , Gene Expression Regulation, Enzymologic/drug effects , Hematopoietic Stem Cells/enzymology , Mutation , Telomerase/biosynthesis , Androgen Antagonists/pharmacology , Androgen Receptor Antagonists , Androgens/therapeutic use , Anemia, Aplastic/drug therapy , Anemia, Aplastic/enzymology , Anemia, Aplastic/genetics , Aromatase Inhibitors/pharmacology , Dyskeratosis Congenita/drug therapy , Dyskeratosis Congenita/enzymology , Dyskeratosis Congenita/genetics , Estradiol/therapeutic use , Estrogen Antagonists/pharmacology , Estrogen Receptor alpha , Estrogens/therapeutic use , Female , Flutamide/pharmacology , Hematopoietic Stem Cells/pathology , Heterozygote , Humans , Letrozole , Lymphocytes/enzymology , Male , Nitriles/pharmacology , Receptors, Androgen/metabolism , Tamoxifen/pharmacology , Telomerase/genetics , Triazoles/pharmacologyABSTRACT
Shwachman-Diamond syndrome (SDS; OMIM 260400), an inherited bone marrow failure syndrome, is caused by mutations in both alleles of the SBDS gene, which encodes a protein of unknown function. Here we report heterozygosity for the 258 + 2 T>C SBDS gene mutation previously identified in SDS patients in 4 of 91 patients with apparently acquired aplastic anemia (AA) but not in 276 ethnically matched controls (Fisher exact test, P < .004). Affected patients were young and had a poor outcome; they had reduced SBDS expression but no evidence of the pancreatic exocrine failure or skeletal abnormalities typical of SDS. Length of telomeres in granulocytes of SBDS heterozygous patients was short for their age, and in SDS patients with both SBDS alleles affected further analyzed, granulocytes' telomeres were even shorter, correlating in length with SBDS expression. Higher heterogeneity in telomere length also was observed in SDS patients. Telomerase activity of SBDS-deficient patients' lymphocytes was comparable with controls, and no physical interaction between SBDS protein and telomerase complex components (TERT or TERC) was established. We propose that heterozygosity for the 258 + 2 T>C SBDS mutation predisposes to AA by accelerating telomere shortening of leukocytes via a telomerase-independent mechanism.
