Transcriptional effects of 1,25 dihydroxyvitamin D3 physiological and supra-physiological concentrations in breast cancer organotypic culture

Background Vitamin D transcriptional effects were linked to tumor growth control, however, the hormone targets were determined in cell cultures exposed to supra physiological concentrations of 1,25(OH)2D3 (50-100nM). Our aim was to evaluate the transcriptional effects of 1,25(OH)2D3 in a more physiological model of breast cancer, consisting of fresh tumor slices exposed to 1,25(OH)2D3 at concentrations that can be attained in vivo. Methods Tumor samples from post-menopausal breast cancer patients were sliced and cultured for 24 hours with or without 1,25(OH)2D3 0.5nM or 100nM. Gene expression was analyzed by microarray (SAM paired analysis, FDR≤0.1) or RT-qPCR (p≤0.05, Friedman/Wilcoxon test). Expression of candidate genes was then evaluated in mammary epithelial/breast cancer lineages and cancer associated fibroblasts (CAFs), exposed or not to 1,25(OH)2D3 0.5nM, using RT-qPCR, western blot or immunocytochemistry. Results 1,25(OH)2D3 0.5nM or 100nM effects were evaluated in five tumor samples by microarray and seven and 136 genes, respectively, were up-regulated. There was an enrichment of genes containing transcription factor binding sites for the vitamin D receptor (VDR) in samples exposed to 1,25(OH)2D3 near physiological concentration. Genes up-modulated by both 1,25(OH)2D3 concentrations were CYP24A1, DPP4, CA2, EFTUD1, TKTL1, KCNK3. Expression of candidate genes was subsequently evaluated in another 16 samples by RT-qPCR and up-regulation of CYP24A1, DPP4 and CA2 by 1,25(OH)2D3 was confirmed. To evaluate whether the transcripitonal targets of 1,25(OH)2D3 0.5nM were restricted to the epithelial or stromal compartments, gene expression was examined in HB4A, C5.4, SKBR3, MDA-MB231, MCF-7 lineages and CAFs, using RT-qPCR. In epithelial cells, there was a clear induction of CYP24A1, CA2, CD14 and IL1RL1. In fibroblasts, in addition to CYP24A1 induction, there was a trend towards up-regulation of CA2, IL1RL1, and DPP4. A higher protein expression of CD14 in epithelial cells and CA2 and DPP4 in CAFs exposed to 1,25(OH)2D3 0.5nM was detected. Conclusions In breast cancer specimens a short period of 1,25(OH)2D3 exposure at near physiological concentration modestly activates the hormone transcriptional pathway. Induction of CYP24A1, CA2, DPP4, IL1RL1 expression appears to reflect 1,25(OH)2D3 effects in epithelial as well as stromal cells, however, induction of CD14 expression is likely restricted to the epithelial compartment.

1,25-Dihydroxycholecalciferol with low-calcium diet reduces acute rejection in rat lung allotransplantation

The effect of 1,25-dihydroxycholecalciferol (calcitriol, vitamin D3) with a low-calcium diet on the acute lung allograft rejection in a rat unilateral left lung transplantation model was evaluated.

Interleukin-17A stimulates granulocyte-macrophage colony-stimulating factor release by murine osteoblasts in the presence of 1,25-dihydroxyvitamin D(3) and inhibits murine osteoclast development in vitro

OBJECTIVE To investigate the effects of interleukin-17A (IL-17A) on osteoclastogenesis in vitro. METHODS Bone marrow cells (BMCs) were isolated from the excised tibia and femora of wild-type C57BL/6J mice, and osteoblasts were obtained by sequential digestion of the calvariae of ddY, C57BL/6J, and granulocyte-macrophage colony-stimulating factor-knockout (GM-CSF(-/-)) mice. Monocultures of BMCs or cocultures of BMCs and osteoblasts were supplemented with or without 1,25-dihydroxyvitamin D(3)(1,25[OH](2)D(3)), recombinant human macrophage colony-stimulating factor (M-CSF), RANKL, and IL-17A. After 5-6 days, the cultures were fixed with 4% paraformaldehyde and subsequently stained for the osteoclast marker enzyme tartrate-resistant acid phosphatase (TRAP). Osteoprotegerin (OPG) and GM-CSF expression were measured by enzyme-linked immunosorbent assay, and transcripts for RANK and RANKL were detected by real-time polymerase chain reaction. RESULTS In both culture systems, IL-17A alone did not affect the development of osteoclasts. However, the addition of IL-17A plus 1,25(OH)(2)D(3) to cocultures inhibited early osteoclast development within the first 3 days of culture and induced release of GM-CSF into the culture supernatants. Furthermore, in cocultures of GM-CSF(-/-) mouse osteoblasts and wild-type mouse BMCs, IL-17A did not affect osteoclast development, corroborating the role of GM-CSF as the mediator of the observed inhibition of osteoclastogenesis by IL-17A. CONCLUSION These findings suggest that IL-17A interferes with the differentiation of osteoclast precursors by inducing the release of GM-CSF from osteoblasts.

PTH and 1.25 vitamin D response to a low-calcium diet is associated with bone mineral density in renal stone formers

BACKGROUND: Renal calcium stones and hypercalciuria are associated with a reduced bone mineral density (BMD). Therefore, the effect of changes in calcium homeostasis is of interest for both stones and bones. We hypothesized that the response of calciuria, parathyroid hormone (PTH) and 1.25 vitamin D to changes in dietary calcium might be related to BMD. METHODS: A single-centre prospective interventional study of 94 hyper- and non-hypercalciuric calcium stone formers consecutively retrieved from our stone clinic. The patients were investigated on a free-choice diet, a low-calcium diet, while fasting and after an oral calcium load. Patient groups were defined according to lumbar BMD (z-score) obtained by dual X-ray absorptiometry (group 1: z-score <-0.5, n = 30; group 2: z-score -0.5-0.5, n = 36; group 3: z-score >0.5, n = 28). The effect of the dietary interventions on calciuria, 1.25 vitamin D and PTH in relation to BMD was measured. RESULTS: An inverse relationship between BMD and calciuria was observed on all four calcium intakes (P = 0.009). On a free-choice diet, 1.25 vitamin D and PTH levels were identical in the three patient groups. However, the relative responses of 1.25 vitamin D and PTH to the low-calcium diet were opposite in the three groups with the highest increase of 1.25 vitamin D in group 1 and the lowest in group 3, whereas PTH increase was most pronounced in group 3 and least in group 1. CONCLUSION: Calcium stone formers with a low lumbar BMD exhibit a blunted response of PTH release and an apparently overshooting production of 1.25 vitamin D following a low-calcium diet.

1,25-(OH)2-16ene-23yne-D3 reduces secondary hyperparathyroidism in uremic rats with little calcemic effect

To compare the effects of vitamin D analogs versus calcitriol on serum levels of Ca, P and parathyroid hormone (PTH). A compound better than calcitriol should increase the Ca x P product less than calcitriol for an equivalent decrease in PTH levels.

Geological map of HANSSONHORNA, Heimefrontfjella, Antarctica (Scale 1:25,000)

Topographic data of this geological map were obtained through stereoscopic aerial photo interpretation. The photogrammetric photo flights were undertaken in 1986 by the Institut für Angewandte Geodäsie, Frankfurt. Horizontal ground control points required for aerial photo interpretation were determined by means of Doppler satellite observation during the 2nd German Neuschwabenland Expedition 1985/86. Vertical ground control points were taken from unpublished map drafts at 1:100 000 scale by Norsk Polarinstitutt, Oslo. The elevation above mean sea level was transferred to Heimefrontfjella barometrically. For this reason assertions concerning the absolute elevation (referred to sea level) are uncertain. Contours and spot heights presented on the map were obtained from the photogrammetric evaluation of the photography taken in 1986; relative elevation data (hight differences) are accurate to approximately ±10 m.

Geological map of RYGHNUTEN, Heimefrontfjella, Antarctica (Scale 1:25,000)

Topographic data of this geological map were obtained through stereoscopic aerial photo interpretation. The photogrammetric photo flights were undertaken in 1986 by the Institut für Angewandte Geodäsie, Frankfurt. Horizontal ground control points required for aerial photo interpretation were determined by means of Doppler satellite observation during the 2nd German Neuschwabenland Expedition 1985/86. Vertical ground control points were taken from unpublished map drafts at 1:100 000 scale by Norsk Polarinstitutt, Oslo. The elevation above mean sea level was transferred to Heimefrontfjella barometrically. For this reason assertions concerning the absolute elevation (referred to sea level) are uncertain. Contours and spot heights presented on the map were obtained from the photogrammetric evaluation of the photography taken in 1986; relative elevation data (hight differences) are accurate to approximately ±10 m.

Geological map of WORSFOLDFJELLET, Heimefrontfjella, Antarctica (Scale 1:25,000)

Topographic data of this geological map were obtained through stereoscopic aerial photo interpretation. The photogrammetric photo flights were undertaken in 1986 by the Institut für Angewandte Geodäsie, Frankfurt. Horizontal ground control points required for aerial photo interpretation were determined by means of Doppler satellite observation during the 2nd German Neuschwabenland Expedition 1985/86. Vertical ground control points were taken from unpublished map drafts at 1:100 000 scale by Norsk Polarinstitutt, Oslo. The elevation above mean sea level was transferred to Heimefrontfjella barometrically. For this reason assertions concerning the absolute elevation (referred to sea level) are uncertain. Contours and spot heights presented on the map were obtained from the photogrammetric evaluation of the photography taken in 1986; relative elevation data (height differences) are accurate to approximately ±10 m.

Geological map of JUCKESKAMMEN, Heimefrontfjella, Antarctica (Scale 1:25,000)

Topographic data of this geological map were obtained through stereoscopic aerial photo interpretation. The photogrammetric photo flights were undertaken in 1986 by the Institut für Angewandte Geodäsie, Frankfurt. Horizontal ground control points required for aerial photo interpretation were determined by means of Doppler satellite observation during the 2nd German Neuschwabenland Expedition 1985/86. Vertical ground control points were taken from unpublished map drafts at 1:100 000 scale by Norsk Polarinstitutt, Oslo. The elevation above mean sea level was transferred to Heimefrontfjella barometrically. For this reason assertions concerning the absolute elevation (referred to sea level) are uncertain. Contours and spot heights presented on the map were obtained from the photogrammetric evaluation of the photography taken in 1986; relative elevation data (hight differences) are accurate to approximately ±10 m.

Geological map of GRAMKROKEN, Heimefrontfjella, Antarctica (Scale 1:25,000)

Topographic data of this geological map were obtained through stereoscopic aerial photo interpretation. The photogrammetric photo flights were undertaken in 1986 by the Institut für Angewandte Geodäsie, Frankfurt. Horizontal ground control points required for aerial photo interpretation were determined by means of Doppler satellite observation during the 2nd German Neuschwabenland Expedition 1985/86. Vertical ground control points were taken from unpublished map drafts at 1:100 000 scale by Norsk Polarinstitutt, Oslo. The elevation above mean sea level was transferred to Heimefrontfjella barometrically. For this reason assertions concerning the absolute elevation (referred to sea level) are uncertain. Contours and spot heights presented on the map were obtained from the photogrammetric evaluation of the photography taken in 1986; relative elevation data (hight differences) are accurate to approximately ±10 m.

Geological map of KVITEBOTNEN, Heimefrontfjella, Antarctica (Scale 1:25,000)

Topographic data of this geological map were obtained through stereoscopic aerial photo interpretation. The photogrammetric photo flights were undertaken in 1986 by the Institut für Angewandte Geodäsie, Frankfurt. Horizontal ground control points required for aerial photo interpretation were determined by means of Doppler satellite observation during the 2nd German Neuschwabenland Expedition 1985/86. Vertical ground control points were taken from unpublished map drafts at 1:100 000 scale by Norsk Polarinstitutt, Oslo. The elevation above mean sea level was transferred to Heimefrontfjella barometrically. For this reason assertions concerning the absolute elevation (referred to sea level) are uncertain. Contours and spot heights presented on the map were obtained from the photogrammetric evaluation of the photography taken in 1986; relative elevation data (hight differences) are accurate to approximately ±10 m.