Calcitriol regulates immune genes CD14 and CD180 to modulate LPS responses in human trophoblasts

We assessed the response of primary cultures of placental villous mononucleated trophoblasts and multinucleated syncytiotrophoblast to calcitriol, the most biologically active form of vitamin D. Whole-genome microarray data showed that calcitriol modulates the expression of many genes in trophoblasts within 6 hours of exposure and RT-qPCR revealed similar responses in cytotrophoblasts, syncytiotrophoblasts and villous explants. Both cytotrophoblasts and syncytiotrophoblasts expressed genes for the vitamin D receptor, for LRP2 and CUBN that mediate internalization of calcidiol, forCYP27B1that encodes the enzyme that converts calcidiol into active calcitriol, and forCYP24A1that encodes the enzyme that modifies calcitriol and calcidiol to inactive calcitetrol. Notably, we found an inverse effect of calcitriol on expression of CD14 and CD180/RP105, proteins that differentially regulate toll-like receptor 4-mediated immune responses. Supported by gene ontology analysis, we tested the hypothesis that CD14 and CD180 modulate the inflammatory response of syncytiotrophoblast to bacterial lipopolysaccharide (LPS). These cells showed a robust response to a wide range of LPS concentrations, with induction of active NF-κB and increased secretion of IL-6 and IL-8. SiRNA-mediated knockdown ofCD14reduced the secretion of IL-6 and IL-8 in response to LPS. Collectively, our data showed that calcitriol has a rapid and widespread effect on villous trophoblast gene expression in general, and a specific effect on the innate immune response by syncytiotrophoblast.

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Key Vitamin D Target Genes with Functions in the Immune System

Nutrients ◽

10.3390/nu12041140 ◽

2020 ◽

Vol 12 (4) ◽

pp. 1140 ◽

Cited By ~ 6

Author(s):

Oona Koivisto ◽

Andrea Hanel ◽

Carsten Carlberg

Keyword(s):

Vitamin D ◽

Adaptive Immunity ◽

Vitamin D3 ◽

Target Genes ◽

Mononuclear Cells ◽

Active Form ◽

Biologically Active ◽

Monocytic Cell ◽

Monocytic Cell Line ◽

Innate And Adaptive Immunity

The biologically active form of vitamin D3, 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3), modulates innate and adaptive immunity via genes regulated by the transcription factor vitamin D receptor (VDR). In order to identify the key vitamin D target genes involved in these processes, transcriptome-wide datasets were compared, which were obtained from a human monocytic cell line (THP-1) and peripheral blood mononuclear cells (PBMCs) treated in vitro by 1,25(OH)2D3, filtered using different approaches, as well as from PBMCs of individuals supplemented with a vitamin D3 bolus. The led to the genes ACVRL1, CAMP, CD14, CD93, CEBPB, FN1, MAPK13, NINJ1, LILRB4, LRRC25, SEMA6B, SRGN, THBD, THEMIS2 and TREM1. Public epigenome- and transcriptome-wide data from THP-1 cells were used to characterize these genes based on the level of their VDR-driven enhancers as well as the level of the dynamics of their mRNA production. Both types of datasets allowed the categorization of the vitamin D target genes into three groups according to their role in (i) acute response to infection, (ii) infection in general and (iii) autoimmunity. In conclusion, 15 genes were identified as major mediators of the action of vitamin D in innate and adaptive immunity and their individual functions are explained based on different gene regulatory scenarios.

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Altered regulation of cytosolic Ca2+ concentration in dendritic cells from klotho hypomorphic mice

AJP Cell Physiology ◽

10.1152/ajpcell.00355.2012 ◽

2013 ◽

Vol 305 (1) ◽

pp. C70-C77 ◽

Cited By ~ 10

Author(s):

Ekaterina Shumilina ◽

Meerim K. Nurbaeva ◽

Wenting Yang ◽

Evi Schmid ◽

Kalina Szteyn ◽

...

Keyword(s):

Bone Marrow ◽

Vitamin D ◽

Dendritic Cells ◽

Active Form ◽

Biologically Active ◽

Deficient Diet ◽

Dihydroxyvitamin D3 ◽

Mhc Ii ◽

Macrophage Colony ◽

Antigen Presenting

The function of dendritic cells (DCs), antigen-presenting cells regulating naïve T-cells, is regulated by cytosolic Ca2+ concentration ([Ca2+]i). [Ca2+]i is increased by store-operated Ca2+ entry and decreased by K+-independent (NCX) and K+-dependent (NCKX) Na+/Ca2+ exchangers. NCKX exchangers are stimulated by immunosuppressive 1,25-dihydroxyvitamin D3 [1,25(OH)2D3], the biologically active form of vitamin D. Formation of 1,25(OH)2D3 is inhibited by the antiaging protein Klotho. Thus 1,25(OH)2D3 plasma levels are excessive in Klotho-deficient mice ( klotho hm). The present study explored whether Klotho deficiency modifies [Ca2+]i regulation in DCs. DCs were isolated from the bone marrow of klotho hm mice and wild-type mice ( klotho+/+) and cultured for 7–9 days with granulocyte-macrophage colony-stimulating factor. According to major histocompatibility complex II (MHC II) and CD86 expression, differentiation and lipopolysaccharide (LPS)-induced maturation were similar in klotho hm DCs and klotho+/+ DCs. However, NCKX1 membrane abundance and NCX/NCKX-activity were significantly enhanced in klotho hm DCs. The [Ca2+]i increase upon acute application of LPS (1 μg/ml) was significantly lower in klotho hm DCs than in klotho+/+ DCs, a difference reversed by the NCKX blocker 3′,4′-dichlorobenzamyl (DBZ; 10 μM). CCL21-dependent migration was significantly less in klotho hm DCs than in klotho+/+ DCs but could be restored by DBZ. NCKX activity was enhanced by pretreatment of klotho+/+ DC precursors with 1,25(OH)2D3 the first 2 days after isolation from bone marrow. Feeding klotho hm mice a vitamin D-deficient diet decreased NCKX activity, augmented LPS-induced increase of [Ca2+]i, and enhanced migration of klotho hm DCs, thus dissipating the differences between klotho hm DCs and klotho+/+ DCs. In conclusion, Klotho deficiency upregulates NCKX1 membrane abundance and Na+/Ca2+-exchange activity, thus blunting the increase of [Ca2+]i following LPS exposure and CCL21-mediated migration. The effects are in large part due to excessive 1,25(OH)2D3 formation.

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Vitamin D Treatment Sequence Is Critical for Transcriptome Modulation of Immune Challenged Primary Human Cells

Frontiers in Immunology ◽

10.3389/fimmu.2021.754056 ◽

2021 ◽

Vol 12 ◽

Author(s):

Henna-Riikka Malmberg ◽

Andrea Hanel ◽

Mari Taipale ◽

Sami Heikkinen ◽

Carsten Carlberg

Keyword(s):

Vitamin D ◽

Mononuclear Cells ◽

Fungal Infections ◽

Human Peripheral Blood ◽

Active Form ◽

Biologically Active ◽

Treatment Sequence ◽

Peripheral Blood Mononuclear ◽

Lps Challenge ◽

Immune Challenges

Microbe-associated molecular patterns, such as lipopolysaccharide (LPS) and β-glucan (BG), are surrogates of immune challenges like bacterial and fungal infections, respectively. The biologically active form of vitamin D, 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3), supports the immune system in its fight against infections. This study investigated significant and prominent changes of the transcriptome of human peripheral blood mononuclear cells that immediately after isolation are exposed to 1,25(OH)2D3-modulated immune challenges over a time frame of 24-48 h. In this in vitro study design, most LPS and BG responsive genes are downregulated and their counts are drastically reduced when cells are treated 24 h after, 24 h before or in parallel with 1,25(OH)2D3. Interestingly, only a 1,25(OH)2D3 pre-treatment of the LPS challenge results in a majority of upregulated genes. Based on transcriptome-wide data both immune challenges display characteristic differences in responsive genes and their associated pathways, to which the actions of 1,25(OH)2D3 often oppose. The joined BG/1,25(OH)2D3 response is less sensitive to treatment sequence than that of LPS/1,25(OH)2D3. In conclusion, the functional consequences of immune challenges are significantly modulated by 1,25(OH)2D3 but largely depend on treatment sequence. This may suggest that a sufficient vitamin D status before an infection is more important than vitamin D supplementation afterwards.

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Vitamin D as a regulator of steroidogenic enzymes

F1000Research ◽

10.12688/f1000research.4714.1 ◽

2014 ◽

Vol 3 ◽

pp. 155 ◽

Cited By ~ 5

Author(s):

Johan Lundqvist

Keyword(s):

Vitamin D ◽

Steroid Hormones ◽

Sex Hormones ◽

Active Form ◽

Animal Experiments ◽

Physiological Role ◽

Steroidogenic Enzymes ◽

Adrenal Steroid ◽

Wide Range ◽

Sexual Characteristics

During the last decades, the outlook on vitamin D has widened, from being a vitamin solely involved in bone metabolism and calcium homeostasis, to being a multifunctional hormone known to affect a broad range of physiological processes. The aim of this review is to summarize the research on vitamin D as a regulator of steroidogenic enzymes. Steroid hormones exert a wide range of physiological responses, including functions in the immune system, protein and carbohydrate metabolism, water and salt balance, reproductive system and development of sexual characteristics. The balance of sex hormones is also of importance in the context of breast and prostate cancer. Steroid hormones are synthesized in steroidogenic tissues such as the adrenal cortex, breast, ovaries, prostate and testis, either from cholesterol or from steroidogenic precursors secreted from other steroidogenic tissues. The hormonally active form of vitamin D has been reported to act as a regulator of a number of enzymes involved in the regulation of steroid hormon production, and thereby the production of both adrenal steroid hormones and sex hormones. The research reviewed in the article has in large part been performed in cell culture based experiments and laboratory animal experiments, and the physiological role of the vitamin D mediated regulation of steroidogenic enzyme need to be further investigated.

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Vitamin D Enhances Radiosensitivity of Colorectal Cancer by Reversing Epithelial-Mesenchymal Transition

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.684855 ◽

2021 ◽

Vol 9 ◽

Author(s):

Xinyue Yu ◽

Qian Wang ◽

Baocai Liu ◽

Ning Zhang ◽

Guanghui Cheng

Keyword(s):

Colorectal Cancer ◽

Vitamin D ◽

Epithelial Mesenchymal Transition ◽

Active Form ◽

Plasminogen Activator Inhibitor ◽

Biologically Active ◽

Plasminogen Activator Inhibitor 1 ◽

Mesenchymal Transition

Colorectal cancer (CRC) is often resistant to conventional therapies. Previous studies have reported the anticancer effects of vitamin D in several cancers, its role in radiotherapy (RT) remains unknown. We found that 1α, 25-dihydroxyvitamin D3 (VD3), the biologically active form of vitamin D, had antitumor effect on CRC and sensitized CRC cells to ionizing radiation (IR). VD3 demonstrated synergistic effect in combination with IR, which were detected by colony formation and cell proliferation assay. Radiosensitivity restoration induced by VD3 was associated with a series of phenotypes, including apoptosis, autophagy, and epithelial-mesenchymal transition (EMT). Using proteomics, “regulation of cell migration” and “cadherin” were found to be obviously enriched GO terms. Moreover, cystatin D and plasminogen activator inhibitor-1 (PAI-1), the differentially expressed proteins, were associated with EMT. Next, we confirmed the contributions of these two genes in enhancing IR sensitivity of CRC cells upon inhibition of EMT. As determined by proteomics, the mechanism underlying such sensitivity involved partially block of JAK/STAT3 signaling pathway. Furthermore, VD3 also elicited sensitization to RT in xenograft CRC models without additional toxicity. Our study revealed that VD3 was able to act in synergy with IR both in vitro and in vivo and could also confer radiosensitivity by regulating EMT, thereby providing a novel insight for elevating the efficacy of therapeutic regimens.

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Impact of Epigenetics on Complications of Fanconi Anemia: The Role of Vitamin D-Modulated Immunity

Nutrients ◽

10.3390/nu12051355 ◽

2020 ◽

Vol 12 (5) ◽

pp. 1355

Author(s):

Eunike Velleuer ◽

Carsten Carlberg

Keyword(s):

Vitamin D ◽

Fanconi Anemia ◽

Bone Marrow Failure ◽

Active Form ◽

Cellular Growth ◽

Biologically Active ◽

Fine Tuning ◽

Dihydroxyvitamin D3 ◽

Increased Risk ◽

The Individual

Fanconi anemia (FA) is a rare disorder with the clinical characteristics of (i) specific malformations at birth, (ii) progressive bone marrow failure already during early childhood and (iii) dramatically increased risk of developing cancer in early age, such as acute myeloid leukemia and squamous cell carcinoma. Patients with FA show DNA fragility due to a defect in the DNA repair machinery based on predominately recessive mutations in 23 genes. Interestingly, patients originating from the same family and sharing an identical mutation, frequently show significant differences in their clinical presentation. This implies that epigenetics plays an important role in the manifestation of the disease. The biologically active form of vitamin D, 1α,25-dihydroxyvitamin D3 controls cellular growth, differentiation and apoptosis via the modulation of the immune system. The nuclear hormone activates the transcription factor vitamin D receptor that affects, via fine-tuning of the epigenome, the transcription of >1000 human genes. In this review, we discuss that changes in the epigenome, in particular in immune cells, may be central for the clinical manifestation of FA. These epigenetic changes can be modulated by vitamin D suggesting that the individual FA patient’s vitamin D status and responsiveness are of critical importance for disease progression.

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Vitamin D Deficiency as It Relates to Oral Immunity and Chronic Periodontitis

International Journal of Dentistry ◽

10.1155/2018/7315797 ◽

2018 ◽

Vol 2018 ◽

pp. 1-9 ◽

Cited By ~ 13

Author(s):

R. A. G. Khammissa ◽

R. Ballyram ◽

Y. Jadwat ◽

J. Fourie ◽

J. Lemmer ◽

...

Keyword(s):

Vitamin D ◽

Serum Level ◽

Chronic Periodontitis ◽

Active Form ◽

Antibacterial Properties ◽

Vitamin D Supplementation ◽

Biologically Active ◽

Periodontal Treatment ◽

Periodontal Health ◽

Cross Sectional

The biologically active form of vitamin D, 1,25 dihydroxyvitamin D (1,25(OH)2D) and its receptor, the vitamin D receptor (VDR), play roles in maintaining oral immunity and the integrity of the periodontium. Results of observational cross-sectional clinical studies investigating the association between vitamin D serum level and the incidence and severity of chronic periodontitis indicate that, perhaps owing to the immunomodulatory, anti-inflammatory, and antibacterial properties of 1,25(OH)2D/VDR signalling, a sufficient serum level of vitamin D is necessary for the maintenance of periodontal health. In cases of established chronic periodontitis, vitamin D supplementation is associated with reduction in the severity of periodontitis. As cross-sectional studies provide only weak evidence for any causal association and therefore are of questionable value, either longitudinal cohort studies, case controlled studies, or randomized control trials are needed to determine whether or not deficiency of vitamin D is a risk factor for chronic periodontitis, and whether or not vitamin D supplementation adjunctive to standard periodontal treatment is in any way beneficial. In this article, we discuss the relationship between vitamin D, oral immunity and periodontal disease and review the rationale for using vitamin D supplementation to help maintain periodontal health and as an adjunct to standard periodontal treatment.

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Vitamin D and Lumisterol Hydroxyderivatives Can Act on Liver X Receptors (LXRs)

Journal of the Endocrine Society ◽

10.1210/jendso/bvab048.1671 ◽

2021 ◽

Vol 5 (Supplement_1) ◽

pp. A820-A820

Author(s):

Andrzej Slominski ◽

Tae-Kang Kim ◽

Shariq Qayyum ◽

Yuwei Song ◽

Zorica Janjetovic ◽

...

Keyword(s):

Vitamin D ◽

Target Genes ◽

Active Form ◽

Human Keratinocytes ◽

Major Product ◽

Dermal Fibroblasts ◽

Biologically Active ◽

Liver X Receptors ◽

X Receptors ◽

Dynamics Simulations

Abstract New pathways of vitamin D3 (D3) activation initiated by CYP11A1 and involving other CYPs have been discovered. At least 15 hydroxyderivatives, including 20(OH)D3 as the major product, are generated by these pathways (1,2) with some being present in human serum, epidermis, and pig adrenals. CYP11A1 can also metabolize 7-dehydrocholesterol to produce 7-dehydropregnenolone, which can be further modified by steroidogenic enzymes generating Δ7-steroids (1,2). Lastly, CYP11A1 and CYP27A1 act on lumisterol (L3) producing at least 9 biologically active derivatives (1,2). Thus, new pathways generating a large number of biologically active secosteroids and lumisterol-derivatives have now been described. These compounds interact with the vitamin D receptor (VDR), retinoic acid receptors (RORs) α and γ, and the aryl hydrocarbon receptor (AhR)(1). These findings challenge dogmas that lumisterol is biologically inactive and that 1,25(OH)2D3 is the only active form of D3 exerting its effects exclusively through interaction with the VDR. In view of the above and since liver X receptors (LXRs) can be activated by oxysterols, we investigated the interactions of novel products of L3 and D3 metabolism with LXRs. Molecular docking, using crystal structures of the ligand binding domains (LBDs) of LXRα and β, revealed high docking scores for L3 and D3 hydroxymetabolites, like those of the natural ligands, predicting good receptor binding. RNA sequencing of murine dermal fibroblasts stimulated with D3-hydroxyderivatives revealed LXR as the second major nuclear receptor signaling pathway for several D3-hydroxyderivatives, including 1,25(OH)2D3. The involvement of LXRs was validated by the induction of several genes downstream of LXR. Furthermore, L3 and D3-hydroxyderivatives activated an LXR-response element (LXRE)-driven reporter in CHO cells and human keratinocytes. For keratinocytes, enhanced expression of LXR target genes was also observed supporting the involvement of LXR. Importantly, L3 and D3 derivatives showed high affinity binding to the LBD of the LXRα and β in LanthaScreen TR-FRET LXRα and β coactivator assays. The majority of metabolites functioned as LXRα/β agonists; however, 1,20,25(OH)3D3, 1,25(OH)2D3, 1,20(OH)2D3 and 25(OH)D3 acted as inverse agonists of LXRα, but as agonists of LXRβ. Molecular dynamics simulations performed for selected compounds, including 1,25(OH)2D3, 1,20(OH)2D3, 25(OH)D3, 20(OH)D3, 20(OH)L3 and 20,22(OH)2L3, showed overlapping and different interactions with LXRs. Identification of D3 and L3 derivatives as ligands for LXRs changes the accepted paradigms on their biological role and mechanism of action. 1. Cell Biochem Biophys. 2020;78(2):165-180. 2. J Steroid Biochem Mol Biol. 2019;186:4-21.

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Is There Pandemic Vitamin D Deficiency in the Black Population? A Review of Evidence

The Open Nutrition Journal ◽

10.2174/1876396001509010005 ◽

2015 ◽

Vol 9 (1) ◽

pp. 5-11 ◽

Cited By ~ 4

Author(s):

Ria S. Roberts ◽

Fafa Huberta Koudoro ◽

Mark S. Elliott ◽

Zhiyong Han

Keyword(s):

Vitamin D ◽

Vitamin D Deficiency ◽

Active Form ◽

Serum Vitamin ◽

Black Population ◽

Biologically Active ◽

Total Serum ◽

Vitamin D Status ◽

Mineral Density ◽

Serum Vitamin D

Although 1,25-dihydroxyvitamin D [1,25(OH)2D] is the biologically active form of vitamin D, measurement of the total serum 25-hydroxyvitamin D [25(OH)D] level is the gold standard used to define vitamin D status. Currently, it is widely accepted that serum 25 (OH) D levels below 20 ng/ml defines vitamin D deficiency. According to this definition, there appears to be pandemic vitamin D deficiency in the Black population. However, there is no evidence of higher-than-normal rates of common complications and symptomology of true vitamin D deficiency in the Black population. What is going on? We researched the MEDLINE databases to find studies, from 1967 to present, that directly compare between Blacks and Caucasians the following: serum vitamin D level, serum calcium level, serum parathyroid hormone level, bone mineral density and health, and non-skeletal risks associated with vitamin D deficiency. The available studies consistently show that Blacks tend to have serum 25(OH)D levels in the deficient range while their serum 1,25(OH)D level is similar to, if not even slightly higher than that of Caucasians, and that the serum Ca2+ level in Blacks is virtually identical to that in Caucasians. Therefore, it appears that the serum 25(OH)D level is not the best marker of vitamin D sufficiency or deficiency in Blacks. In the future, clinical evaluation of the vitamin D status in the Black population needs to consider other serum biomarkers such as 1,25(OH)2D and/or bioavailable 25(OH)D.

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Common and personal target genes of the micronutrient vitamin D in primary immune cells from human peripheral blood

Scientific Reports ◽

10.1038/s41598-020-78288-0 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Andrea Hanel ◽

Antonio Neme ◽

Marjo Malinen ◽

Emmi Hämäläinen ◽

Henna-Riikka Malmberg ◽

...

Keyword(s):

Vitamin D ◽

Immune System ◽

Immune Cells ◽

Peripheral Blood ◽

Target Genes ◽

Mononuclear Cells ◽

Human Peripheral Blood ◽

Active Form ◽

Biologically Active ◽

Expression Change

AbstractVitamin D is essential for the function of the immune system. In this study, we treated peripheral blood mononuclear cells (PBMCs) of healthy adults with the biologically active form of vitamin D3, 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3) using two different approaches: single repeats with PBMCs obtained from a cohort of 12 individuals and personalized analysis based on triplicates of five study participants. This identified 877 (cohort approach) and 3951 (personalized approach) genes that significantly (p < 0.05) changed their expression 24 h after 1,25(OH)2D3 stimulation. From these, 333 and 1232 were classified as supertargets, a third of which were identified as novel. Individuals differed largely in their vitamin D response not only by the magnitude of expression change but also by their personal selection of (super)target genes. Functional analysis of the target genes suggested the overarching role of vitamin D in the regulation of metabolism, proliferation and differentiation, but in particular in the control of functions mediated by the innate and adaptive immune system, such as responses to infectious diseases and chronic inflammatory disorders. In conclusion, immune cells are an important target of vitamin D and common genes may serve as biomarkers for personal responses to the micronutrient.

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