1,25-dihydroxyvitamin D-3 hypersensitivity of osteoclast precursors from patients with Paget's disease

Our previous studies suggested that increased osteoclast formation and acti vity in Paget's disease may be related in part to increased responsiveness of highly purified osteoclast precursors to 1,25-dihydroxyvitamin D-3 [1,25 -(OH)(2)D-3]. However, the basis for this enhanced sensitivity to 1,25-(OH) ,D3 is unclear To address this question, we examined 24-hydroxylase and 1,2 5-(OH)(2)D-3 receptor (VDR) messenger RNA (mRNA) expression during human os teoclast differentiation from normal subjects and patients with Paget's dis ease in response to 1,25-(OH)(2)D-3 as well as VDR content and affinity, Re verse-transcription polymerase chain reaction (RT-PCR) analysis of granuloc yte-macrophage colony-forming unit (GM-CFU), the earliest identifiable oste oclast precursor, derived from patients with Paget's disease demonstrated 2 4-hydroxylase mRNA expression in response to 1,25-(OH)(2)D-3 was induced at concentrations of 1,25-(OH)(2)D-3 that were at least one log less than tha t required for normal GM-CFU, VDR mRNA and VDR protein were detected in bot h immature and more differentiated osteoclast precursors, as well as in ost eoclast-like multinucleated cells (MNCs), However, VDR expression was lower in MNCs than the mononuclear precursor cells, Osteoclast precursors and MN Cs from patients with Paget's disease had levels of VDR expression similar to those of normal subjects but showed increased VDR affinity for 1,25-(OH) (2)D-3. Because the effects of 1,25-(OH)(2)D-3 are in part mediated by indu ction of expression of RANK ligand on marrow stromal cells, which in turn s timulates osteoclast formation, we examined expression of RANK ligand mRNA by marrow stromal cell lines derived from patients with Paget's disease and normal subjects in response to 1,25-(OH)(2)D-3. RT-PCR analysis showed no difference in sensitivity of marrow stromal cells to 1,25-(OH)(2)D-3 from n ormal subjects or patients with Paget's disease although the Paget's stroma l cells expressed increased basal levels of RANK ligand mRNA, These results show that VDR protein is expressed in early and more differentiated osteoc last precursors, that expression levels of VDR decline with osteoclast diff erentiation, and that 1,25-(OH)(2)D-3 has direct effects on osteoclast prec ursors, The enhanced sensitivity to 1,25-(OH)(2)D-3 is an intrinsic propert y of osteoclast precursors from patients with Paget's disease that distingu ishes them from normal osteoclast precursors, Furthermore, our results sugg est that an increased affinity of VDR for 1,25-(OH)(2)D-3 may be responsibl e for the enhanced 1,25-(OH)(2)D-3 sensitivity of osteoclast precursors in patients with Paget's disease compared with normal subjects.