|
1
|
Banerjee A, Ganguly U, Saha S, Chakrabarti S, Saini RV, Rawal RK, Saso L, Chakrabarti S. Vitamin D and immuno-pathology of COVID-19: many interactions but uncertain therapeutic benefits. Expert Rev Anti Infect Ther 2021; 19:1245-1258. [PMID: 33739215 PMCID: PMC8022339 DOI: 10.1080/14787210.2021.1905519] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/06/2021] [Accepted: 03/16/2021] [Indexed: 02/06/2023]
Abstract
Introduction: COVID-19 pandemic has caused huge loss of human lives and extensive socio-economic damages. The immuno-pathology of this disease is neither clearly understood nor there are effective drugs for severe cases of COVID-19. Repurposing of available drugs for the treatment of COVID-19 is imperative.Areas Covered: This review has gathered the evidence from PubMed, Google Scholar, WHO, and other reliable websites on COVID-19 and summarized the existing knowledge of the immuno-pathology of COVID-19. We elucidated how vitamin D through its diverse actions on immune effector cells, epithelial cells, or renin-angiotensin-aldosterone system could have a modulatory role on the pathogenic mechanisms of COVID-19. The epidemiological evidence associating vitamin D deficiency with the severity and incidence of COVID-19 is also presented. However, the evidence of clinical benefit to patients of COVID-19 from randomized controlled trials with vitamin D has not come as yet.Expert opinion: It is now established that fatality of COVID-19 is primarily determined by hyperactivation of the host's innate immune system in response to SARS-CoV-2 invasion, and thus the research on the immuno-modulatory and other roles of vitamin D against viral infections should be pursued vigorously. This would be also useful for future pandemics caused by other novel viruses.
Collapse
Affiliation(s)
- Anindita Banerjee
- Department of Biochemistry, Institute of Post Graduate Medical Education and Research, Kolkata, India
| | - Upasana Ganguly
- Department of Biochemistry & Central Research Cell, M.M. Institute of Medical Sciences and Research, Maharishi Markandeshwar (Deemed to Be University), Mullana, India
| | - Sarama Saha
- Department of Biochemistry, All India Institute of Medical Sciences, Rishikesh, India
| | | | - Reena V Saini
- Department of Biotechnology, M.M Engineering College, Maharishi Markandeshwar (Deemed to Be University), Mullana, India
| | - Ravindra K Rawal
- Department of Chemistry, M.M Engineering College, Maharishi Markandeshwar (Deemed to Be University), Mullana, India
| | - Luciano Saso
- Department of Physiology and Pharmacology “Vittorio Erspamer”, Sapienza University of Rome, Rome, Italy
| | - Sasanka Chakrabarti
- Department of Biochemistry & Central Research Cell, M.M. Institute of Medical Sciences and Research, Maharishi Markandeshwar (Deemed to Be University), Mullana, India
| |
Collapse
|
|
2
|
Pillar S, Amer R. The association between vitamin D and uveitis: A comprehensive review. Surv Ophthalmol 2021; 67:321-330. [PMID: 34343538 DOI: 10.1016/j.survophthal.2021.07.006] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/26/2021] [Revised: 07/26/2021] [Accepted: 07/27/2021] [Indexed: 12/15/2022]
Abstract
Vitamin D plays an important role in both the innate and adaptive immune systems. We review published data on the relationship between uveitis and vitamin D levels or vitamin D-associated gene polymorphisms. A search of the PubMed and Medline databases was conducted to identify relevant articles concerning vitamin D and uveitis. Sixteen studies were included in this review, and the evidence they present, linking low vitamin D levels with uveitis, is compelling. The uveitic entities shown to be modulated by hypovitaminosis D include, but are not limited to, HLA-B27-associated acute anterior uveitis, Vogt-Koyanagi-Harada (VKH) disease, sarcoidosis-associated uveitis, and juvenile idiopathic arthritis-associated uveitis. Specific polymorphisms of vitamin D family genes were found to correlate with uveitis in ankylosing spondylitis, Behçet's disease, VKH, and HLA B27-positive patients. Further understanding of the role of vitamin D, a known regulator of inflammatory processes, in noninfectious uveitis may advance capabilities in the fields of disease prevention and treatment.
Collapse
Affiliation(s)
- Shani Pillar
- Department of Ophthalmology, Hadassah Medical Organization, Hebrew University of Jerusalem, Israel.
| | - Radgonde Amer
- Department of Ophthalmology, Hadassah Medical Organization, Hebrew University of Jerusalem, Israel
| |
Collapse
|
|
3
|
Berardi S, Giardullo L, Corrado A, Cantatore FP. Vitamin D and connective tissue diseases. Inflamm Res 2020; 69:453-462. [PMID: 32172354 DOI: 10.1007/s00011-020-01337-x] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/07/2020] [Revised: 03/04/2020] [Accepted: 03/09/2020] [Indexed: 12/12/2022] Open
Abstract
OBJECTIVE AND DESIGN Recently, many studies have shown that the biologically active form of vitamin D-1,25(OH)2 D-is involved in many biological processes, including immune system modulation, and patients affected by various autoimmune diseases, such as connective tissue diseases (CTD), showed low levels of vitamin D. It is not clear if vitamin D deficiency is involved in the pathogenesis of autoimmune diseases or it is a consequence. MATERIAL We carried out a review of literature to summarize the existing connections between 25-OH vitamin D and CTD. METHODS We searched for articles on PubMed by keywords: vitamin D, connective tissue diseases, systemic lupus erythematosus, Sjogren's syndrome, systemic sclerosis, undifferentiated connective tissue disease. RESULTS The relationship between vitamin D and CTD is still not very clear, despite many studies having been performed and some data suggest a connection between these diseases and 25-OH vitamin D levels. CONCLUSIONS The limitations of the study, such as the heterogeneity of patients, methods used to measure vitamin D serum concentration and other biases, do not lead to unequivocal results to demonstrate a direct link between low vitamin D serum levels and autoimmune diseases. Further studies are needed to resolve conflicting results.
Collapse
Affiliation(s)
- Stefano Berardi
- Department of Medical and Surgical Sciences, Rheumatology Clinic, Ospedali Riuniti Foggia, University of Foggia, Viale Pinto 1, 71121, Foggia, Italy.
| | - Liberato Giardullo
- Department of Medical and Surgical Sciences, Rheumatology Clinic, Ospedali Riuniti Foggia, University of Foggia, Viale Pinto 1, 71121, Foggia, Italy
| | - Addolorata Corrado
- Department of Medical and Surgical Sciences, Rheumatology Clinic, Ospedali Riuniti Foggia, University of Foggia, Viale Pinto 1, 71121, Foggia, Italy
| | - Francesco Paolo Cantatore
- Department of Medical and Surgical Sciences, Rheumatology Clinic, Ospedali Riuniti Foggia, University of Foggia, Viale Pinto 1, 71121, Foggia, Italy
| |
Collapse
|
|
4
|
Czaja AJ. Examining pathogenic concepts of autoimmune hepatitis for cues to future investigations and interventions. World J Gastroenterol 2019; 25:6579-6606. [PMID: 31832000 PMCID: PMC6906207 DOI: 10.3748/wjg.v25.i45.6579] [Citation(s) in RCA: 19] [Impact Index Per Article: 3.2] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/01/2019] [Revised: 11/25/2019] [Accepted: 11/29/2019] [Indexed: 02/06/2023] Open
Abstract
BACKGROUND Multiple pathogenic mechanisms have been implicated in autoimmune hepatitis, but they have not fully explained susceptibility, triggering events, and maintenance or escalation of the disease. Furthermore, they have not identified a critical defect that can be targeted. The goals of this review are to examine the diverse pathogenic mechanisms that have been considered in autoimmune hepatitis, indicate investigational opportunities to validate their contribution, and suggest interventions that might evolve to modify their impact. English abstracts were identified in PubMed by multiple search terms. Full length articles were selected for review, and secondary and tertiary bibliographies were developed. Genetic and epigenetic factors can affect susceptibility by influencing the expression of immune regulatory genes. Thymic dysfunction, possibly related to deficient production of programmed cell death protein-1, can allow autoreactive T cells to escape deletion, and alterations in the intestinal microbiome may help overcome immune tolerance and affect gender bias. Environmental factors may trigger the disease or induce epigenetic changes in gene function. Molecular mimicry, epitope spread, bystander activation, neo-antigen production, lymphocytic polyspecificity, and disturbances in immune inhibitory mechanisms may maintain or escalate the disease. Interventions that modify epigenetic effects on gene expression, alter intestinal dysbiosis, eliminate deleterious environmental factors, and target critical pathogenic mechanisms are therapeutic possibilities that might reduce risk, individualize management, and improve outcome. In conclusion, diverse pathogenic mechanisms have been implicated in autoimmune hepatitis, and they may identify a critical factor or sequence that can be validated and used to direct future management and preventive strategies.
Collapse
Affiliation(s)
- Albert J Czaja
- Division of Gastroenterology and Hepatology, Mayo Clinic College of Medicine and Science, Rochester, MN 55905, United States
| |
Collapse
|
|
5
|
Voo VTF, O'Brien T, Butzkueven H, Monif M. The role of vitamin D and P2X7R in multiple sclerosis. J Neuroimmunol 2019; 330:159-169. [PMID: 30908981 DOI: 10.1016/j.jneuroim.2019.03.004] [Citation(s) in RCA: 4] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/20/2018] [Revised: 02/11/2019] [Accepted: 03/05/2019] [Indexed: 12/13/2022]
Abstract
Multiple sclerosis (MS) is characterized by neuroinflammatory infiltrates and central nervous system demyelination. In the neuroinflammatory foci of MS there is increased expression of a purinergic receptor, P2X7R. Although implicated in the neuroinflammation, the exact role of P2X7R in the context of MS is unclear and forms the basis of this review. In this review, we also introduce the immunopathologies and inflammatory processes in MS, with a focus on P2X7R and the possible immunomodulatory role of vitamin D deficiency in this setting.
Collapse
Affiliation(s)
- Veronica Tsin Fong Voo
- Department of Physiology, The University of Melbourne, Melbourne, Australia; Department of Neuroscience, Monash University, Melbourne, Australia
| | - Terence O'Brien
- Department of Neuroscience, Monash University, Melbourne, Australia; Department of Neurology, Melbourne Health, Melbourne, Australia
| | | | - Mastura Monif
- Department of Physiology, The University of Melbourne, Melbourne, Australia; Department of Neuroscience, Monash University, Melbourne, Australia; Department of Neurology, Melbourne Health, Melbourne, Australia.
| |
Collapse
|
|
6
|
Evolving Role of Vitamin D in Immune-Mediated Disease and Its Implications in Autoimmune Hepatitis. Dig Dis Sci 2019; 64:324-344. [PMID: 30370494 DOI: 10.1007/s10620-018-5351-6] [Citation(s) in RCA: 28] [Impact Index Per Article: 4.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/27/2018] [Accepted: 10/09/2018] [Indexed: 12/11/2022]
Abstract
Vitamin D has immunomodulatory, anti-inflammatory, antioxidant, and anti-fibrotic actions that may impact on the occurrence and outcome of immune-mediated disease. The goals of this review are to describe the nature of these expanded roles, examine the implications of vitamin D deficiency in autoimmune hepatitis, and identify opportunities for future investigation. Abstracts were identified in PubMed by multiple search terms. Full-length articles were selected for review, and secondary and tertiary bibliographies were developed. Vitamin D receptors are expressed on the principal cell populations involved in the innate and adaptive immune responses. Macrophages and dendritic cells can produce 1,25-dihydroxyvitamin D within the microenvironment. This active form of vitamin D can inhibit immune cell proliferation, promote an anti-inflammatory cytokine profile, expand regulatory T cells, enhance glucocorticoid actions, increase glutathione production, and inhibit hepatic stellate cells. Vitamin D deficiency has been commonly present in patients with immune-mediated liver and non-liver diseases, and it has been associated with histological severity, advanced hepatic fibrosis, and non-response to conventional glucocorticoid therapy in autoimmune hepatitis. Vitamin D analogues with high potency, low calcemic effects, and independence from hepatic hydroxylation are possible interventions. In conclusion, vitamin D has properties that could ameliorate immune-mediated disease, and vitamin D deficiency has been a common finding in immune-mediated liver and non-liver diseases, including autoimmune hepatitis. Loss of vitamin D-dependent homeostatic mechanisms may promote disease progression. Vitamin D analogues that are independent of hepatic hydroxylation constitute an investigational opportunity to supplement current management of autoimmune hepatitis.
Collapse
|
|
7
|
The role of vitamin D in hepatic metastases from colorectal cancer. Clin Transl Oncol 2017; 20:259-273. [PMID: 28801869 DOI: 10.1007/s12094-017-1735-x] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/10/2017] [Accepted: 07/30/2017] [Indexed: 02/07/2023]
Abstract
Colorectal cancer (CRC) represents a significant health burden worldwide, comprising approximately 10% of annual cancer cases globally. Hepatic metastases are the most common site of CRC metastasis, and are the leading cause of death in CRC patients. There is strong epidemiologic evidence for an inverse association between vitamin D status and risk of CRC; however, the role of vitamin D in the natural history of liver metastases has not yet been investigated. Several researchers have proposed hallmarks of metastases; crucially, metastases can be blocked by interrupting just one rate-limiting step. Vitamin D status has been implicated in each proposed hallmark of metastasis. The aim of this review is to examine the potential role for vitamin D in reducing the development of hepatic metastases from CRC and outline the candidate mechanisms by which vitamin D may mediate these effects. The results of ongoing randomised intervention trials are eagerly awaited to determine whether addressing vitamin D insufficiency in CRC patients could reduce the occurrence of liver metastases, and the consequent morbidity and mortality.
Collapse
|
|
8
|
|
|
9
|
Lv Y, Yao Q, Ma W, Liu H, Ji J, Li X. Associations of vitamin D deficiency and vitamin D receptor (Cdx-2, Fok I, Bsm I and Taq I) polymorphisms with the risk of primary open-angle glaucoma. BMC Ophthalmol 2016; 16:116. [PMID: 27435453 PMCID: PMC4952063 DOI: 10.1186/s12886-016-0289-y] [Citation(s) in RCA: 27] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/08/2015] [Accepted: 07/05/2016] [Indexed: 11/26/2022] Open
Abstract
BACKGROUND Vitamin D deficiency and vitamin D receptor gene polymorphisms are known to be significantly associated with high myopia. Whether this genetic variant may impact primary open-angle glaucoma is largely unknown. This study investigated whether vitamin D receptor gene polymorphisms are altered in primary open-angle glaucoma subjects carrying the risk allele, and whether vitamin D deficiency is an important factor in the development of glaucoma. METHODS Seventy-three POAG patients and 71 age-matched controls from the Han population were enrolled. Serum levels of 1a, 25-Dihydroxyvitamin D3 were measured by enzyme-linked immunoabsorbent assay. Vitamin D receptor polymorphisms (Cdx-2, Fok I, Bsm I and Taq I) were analyzed using real-time polymerase-chain reaction high resolution melting analysis. RESULTS Serum levels of 1a, 25-Dihydroxyvitamin in primary open-angle glaucoma patients were lower than in age-matched controls. Statistical analysis revealed a significant difference in the allelic frequencies of the BsmI and TaqI genotypes between primary open-angle glaucoma patients and age-matched controls, while other polymorphisms did not show any significant differences. CONCLUSIONS Vitamin D deficiency and the presence of the BsmI 'B' allele and the TaqI 't' allele are relevant risk factors in the development of glaucoma. TRIAL REGISTRATION Clinical Trials.gov: NCT02539745 . The study was registered retrospectively on August 3rd, 2015. The first participant was enrolled on July 4th, 2013.
Collapse
Affiliation(s)
- Yingjuan Lv
- />Department of Glaucoma, Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, The School of Optometry&Ophthalmology, No.251 Fu Kang Road, Nan kai District, Tianjin, 300384 China
| | - Qingbin Yao
- />Department of Histology and Embryology, Tianjin Medical University, No. 22 Qi Xiang Tai Road, He Ping District, Tianjin, 300070 China
| | - Wenjiang Ma
- />Department of Glaucoma, Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, The School of Optometry&Ophthalmology, No.251 Fu Kang Road, Nan kai District, Tianjin, 300384 China
| | - Hua Liu
- />Department of Glaucoma, Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, The School of Optometry&Ophthalmology, No.251 Fu Kang Road, Nan kai District, Tianjin, 300384 China
| | - Jian Ji
- />Department of Glaucoma, Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, The School of Optometry&Ophthalmology, No.251 Fu Kang Road, Nan kai District, Tianjin, 300384 China
| | - Xiaorong Li
- />Department of Glaucoma, Tianjin Medical University Eye Hospital, Tianjin Medical University Eye Institute, The School of Optometry&Ophthalmology, No.251 Fu Kang Road, Nan kai District, Tianjin, 300384 China
| |
Collapse
|
|
10
|
Vitamin D and Alzheimer's Disease: Neurocognition to Therapeutics. Int J Alzheimers Dis 2015; 2015:192747. [PMID: 26351614 PMCID: PMC4553343 DOI: 10.1155/2015/192747] [Citation(s) in RCA: 44] [Impact Index Per Article: 4.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/31/2015] [Accepted: 07/16/2015] [Indexed: 12/21/2022] Open
Abstract
Alzheimer's disease (AD), the major cause of dementia worldwide, is characterized by progressive loss of memory and cognition. The sporadic form of AD accounts for nearly 90% of the patients developing this disease. The last century has witnessed significant research to identify various mechanisms and risk factors contributing to the complex etiopathogenesis of AD by analyzing postmortem AD brains and experimenting with animal and cell culture based models. However, the treatment strategies, as of now, are only symptomatic. Accumulating evidences suggested a significant association between vitamin D deficiency, dementia, and AD. This review encompasses the beneficial role of vitamin D in neurocognition and optimal brain health along with epidemiological evidence of the high prevalence of hypovitaminosis D among aged and AD population. Moreover, disrupted signaling, altered utilization of vitamin D, and polymorphisms of several related genes including vitamin D receptor (VDR) also predispose to AD or AD-like neurodegeneration. This review explores the relationship between this gene-environmental influence and long term vitamin D deficiency as a risk factor for development of sporadic AD along with the role and rationale of therapeutic trials with vitamin D. It is, therefore, urgently warranted to further establish the role of this potentially neuroprotective vitamin in preventing and halting progressive neurodegeneration in AD patients.
Collapse
|
|
11
|
Zhang Y, Leung DYM, Goleva E. Vitamin D enhances glucocorticoid action in human monocytes: involvement of granulocyte-macrophage colony-stimulating factor and mediator complex subunit 14. J Biol Chem 2013; 288:14544-14553. [PMID: 23572530 DOI: 10.1074/jbc.m112.427054] [Citation(s) in RCA: 60] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/16/2022] Open
Abstract
Vitamin D (VitD) is now recognized for its pleiotrophic roles in regulating immune function. VitD interaction with other steroid receptor superfamily receptors in peripheral blood mononuclear cells is poorly understood. In the current study, we demonstrate that VitD enhanced glucocorticoid (GC) responses in human peripheral blood mononuclear cells because it stimulated GC induction of mitogen-activated protein kinase phosphatase-1 (MKP-1) and enhanced GC inhibition of LPS-induced IL-6. These VitD effects were abolished in purified CD14(+) and CD14(-) cells but were recovered in CD14(+) cells co-cultured with CD14(-) cells separated by tissue culture inserts. GM-CSF, found in culture supernatants from CD14(-) cells, was shown to mediate VitD enhancement of GC-induced MKP-1 production in monocytes via increased production of mediator complex subunit 14 (MED14). Recruitment of VitD receptor and MED14, 4.7 kbp upstream of the human MKP-1 gene transcription start site, enhanced binding of glucocorticoid receptor and histone H4 acetylation at the 4.6-kbp glucocorticoid response element of the MKP-1 promoter in the presence of GM-CSF in U937 cells. Knockdown of MED14 abolished VitD-mediated enhancement of GC-induced MKP-1 production. These data demonstrate VitD-mediated stimulation of GC anti-inflammatory effects in human monocytes and identify a role for GM-CSF and MED14 as mediators of this process.
Collapse
Affiliation(s)
- Yong Zhang
- Department of Pediatrics, National Jewish Health, Denver, Colorado 80206
| | - Donald Y M Leung
- Department of Pediatrics, National Jewish Health, Denver, Colorado 80206; Department of Pediatrics, University of Colorado Denver, Aurora, Colorado 80045
| | - Elena Goleva
- Department of Pediatrics, National Jewish Health, Denver, Colorado 80206.
| |
Collapse
|
|
12
|
Zhang Y, Leung DYM, Richers BN, Liu Y, Remigio LK, Riches DW, Goleva E. Vitamin D inhibits monocyte/macrophage proinflammatory cytokine production by targeting MAPK phosphatase-1. JOURNAL OF IMMUNOLOGY (BALTIMORE, MD. : 1950) 2012; 188:2127-35. [PMID: 22301548 PMCID: PMC3368346 DOI: 10.4049/jimmunol.1102412] [Citation(s) in RCA: 622] [Impact Index Per Article: 47.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 12/12/2022]
Abstract
It is estimated that 1 billion people around the world are vitamin D deficient. Vitamin D deficiency has been linked to various inflammatory diseases. However, the mechanism by which vitamin D reduces inflammation remains poorly understood. In this study, we investigated the inhibitory effects of physiologic levels of vitamin D on LPS-stimulated inflammatory response in human blood monocytes and explored potential mechanisms of vitamin D action. We observed that two forms of the vitamin D, 1,25(OH)(2)D(3), and 25(OH)D(3), dose dependently inhibited LPS-induced p38 phosphorylation at physiologic concentrations, IL-6 and TNF-α production by human monocytes. Upon vitamin D treatment, the expression of MAPK phosphatase-1 (MKP-1) was significantly upregulated in human monocytes and murine bone marrow-derived macrophages (BMM). Increased binding of the vitamin D receptor and increased histone H4 acetylation at the identified vitamin D response element of the murine and human MKP-1 promoters were demonstrated. Moreover, in BMM from MKP1(-/-) mice, the inhibition of LPS-induced p38 phosphorylation by vitamin D was completely abolished. Vitamin D inhibition of LPS-induced IL-6 and TNF-α production by BMM from MKP-1(-/-) mice was significantly reduced as compared with wild-type mice. In conclusion, this study identified the upregulation of MKP-1 by vitamin D as a novel pathway by which vitamin D inhibits LPS-induced p38 activation and cytokine production in monocytes/macrophages.
Collapse
Affiliation(s)
- Yong Zhang
- Department of Pediatrics, National Jewish Health, 1400 Jackson Street, Denver, CO 80206 USA
| | - Donald Y. M. Leung
- Department of Pediatrics, National Jewish Health, 1400 Jackson Street, Denver, CO 80206 USA
- Department of Pediatrics, University of Colorado Denver Health Sciences, 13123 East 16 Avenue, Aurora, CO 80045 USA
| | - Brittany N. Richers
- Department of Pediatrics, National Jewish Health, 1400 Jackson Street, Denver, CO 80206 USA
| | - Yusen Liu
- Center for Perinatal Research, The Research Institute at Nationwide Children’s Hospital, Department of Pediatrics, The Ohio State University College of Medicine, 700 Children’s Drive, Columbus, Ohio 43205
| | - Linda K. Remigio
- Department of Pediatrics, National Jewish Health, 1400 Jackson Street, Denver, CO 80206 USA
| | - David W. Riches
- Department of Pediatrics, National Jewish Health, 1400 Jackson Street, Denver, CO 80206 USA
| | - Elena Goleva
- Department of Pediatrics, National Jewish Health, 1400 Jackson Street, Denver, CO 80206 USA
| |
Collapse
|
|
13
|
|
|
14
|
Saramäki A, Banwell CM, Campbell MJ, Carlberg C. Regulation of the human p21(waf1/cip1) gene promoter via multiple binding sites for p53 and the vitamin D3 receptor. Nucleic Acids Res 2006; 34:543-54. [PMID: 16434701 PMCID: PMC1351372 DOI: 10.1093/nar/gkj460] [Citation(s) in RCA: 187] [Impact Index Per Article: 9.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/05/2023] Open
Abstract
The main regulator of the human tumor suppresser gene p21(waf1/cip1) is the transcription factor p53, but more recently it has been suggested to be a primary anti-proliferative target for the nuclear receptor VDR in the presence of its ligand 1alpha,25-dihydroxyvitamin D3 (1alpha,25(OH)2D3). To identify VDR responding regions, we analyzed 20 overlapping regions covering the first 7.1 kb of the p21(waf1/cip1) promoter in MCF-7 human breast cancer cells using chromatin immuno-precipitation assays (ChIP) with antibodies against p53 and VDR. We confirmed two known p53 binding regions at approximate positions -1400 and -2300 and identified a novel site at position -4500. In addition, we found three VDR-associated promoter regions at positions -2300, -4500 and -6900, i.e. two regions showed binding for both p53 and VDR. In silico screening and in vitro binding assays using recombinant and in vitro translated proteins identified five p53 binding sites within the three p53-positive promoter regions and also five 1alpha,25(OH)2D3 response elements within the three VDR-positive regions. Reporter gene assays confirmed the expected responsiveness of the respective promoter regions to the p53 inducer 5-fluorouracil and 1alpha,25(OH)2D3. Moreover, re-ChIP assays confirmed the functionality of the three 1alpha,25(OH)2D3-reponsive promoter regions by monitoring simultaneous occupancy of VDR with the co-activator proteins CBP, SRC-1 and TRAP220. Taken together, we demonstrated that the human p21((waf1/cip1)) gene is a primary 1alpha,25(OH)2D3-responding gene with at least three VDR binding promoter regions, in two of which also p53 co-localizes.
Collapse
Affiliation(s)
- Anna Saramäki
- Department of Biochemistry, University of KuopioFIN-70211 Kuopio, Finland
| | - Claire M. Banwell
- Department of Biochemistry, University of KuopioFIN-70211 Kuopio, Finland
- Division of Medical Sciences, Institute of Biomedical Research, University of Birmingham Medical SchoolEdgbaston, Birmingham, B15 2TH, UK
| | - Moray J. Campbell
- Division of Medical Sciences, Institute of Biomedical Research, University of Birmingham Medical SchoolEdgbaston, Birmingham, B15 2TH, UK
| | - Carsten Carlberg
- Department of Biochemistry, University of KuopioFIN-70211 Kuopio, Finland
- To whom correspondence should be addressed. Tel: +358 17 163062; Fax: +358 17 2811510;
| |
Collapse
|
|
15
|
Väisänen S, Dunlop TW, Sinkkonen L, Frank C, Carlberg C. Spatio-temporal activation of chromatin on the human CYP24 gene promoter in the presence of 1alpha,25-Dihydroxyvitamin D3. J Mol Biol 2005; 350:65-77. [PMID: 15919092 DOI: 10.1016/j.jmb.2005.04.057] [Citation(s) in RCA: 115] [Impact Index Per Article: 5.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/28/2005] [Revised: 04/20/2005] [Accepted: 04/26/2005] [Indexed: 01/29/2023]
Abstract
The vitamin D3 24-hydroxylase gene (CYP24) is one of the most strongly induced genes known. Despite this, its induction by the hormone 1alpha,25-dihydroxyvitamin D3 (1alpha,25OH2D3) has been characterized only partially. Therefore, we monitored the spatio-temporal, 1alpha,25OH2D3-dependent chromatin acetylation status of the human CYP24 promoter by performing chromatin immunoprecipitation (ChIP) assays with antibodies against acetylated histone 4. This was achieved by performing PCR on 25 contiguous genomic regions spanning the first 7.7 kb of the promoter. ChIP assays using antibodies against the 1alpha,25OH2D3 receptor (VDR) revealed that, in addition to the proximal promoter, three novel regions further upstream associated with VDR. Combined in silico/in vitro screening identified in three of the four promoter regions sequences resembling known VDREs and reporter gene assays confirmed the inducibility of these regions by 1alpha,25OH2D3)=. In contrast, the fourth VDR-associated promoter region did not contain any recognizable classical VDRE that could account for the presence of the protein on this region. However, re-ChIP assays monitored on all four promoter regions simultaneous association of VDR with retinoid X receptor, coactivator, mediator and RNA polymerase II proteins. These proteins showed a promoter region-specific association pattern demonstrating the complex choreography of the CYP24 gene promoter activation over 300 minutes. Thus, this study reveals new information concerning the regulation of the CYP24 gene by 1alpha,25OH2D3, and is a demonstration of the simultaneous participation of multiple, structurally diverse response elements in promoter activation in a living cell.
Collapse
Affiliation(s)
- Sami Väisänen
- Department of Biochemistry, University of Kuopio, FIN-70211 Kuopio, Finland
| | | | | | | | | |
Collapse
|
|
16
|
Dunlop TW, Väisänen S, Frank C, Molnár F, Sinkkonen L, Carlberg C. The Human Peroxisome Proliferator-activated Receptor δ Gene is a Primary Target of 1α,25-Dihydroxyvitamin D3 and its Nuclear Receptor. J Mol Biol 2005; 349:248-60. [PMID: 15890193 DOI: 10.1016/j.jmb.2005.03.060] [Citation(s) in RCA: 156] [Impact Index Per Article: 7.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/03/2005] [Revised: 03/15/2005] [Accepted: 03/21/2005] [Indexed: 02/07/2023]
Abstract
Peroxisome proliferator-activated receptor (PPAR) delta is the most widely expressed member of the PPAR family of nuclear receptor fatty acid sensors. Real-time PCR analysis of breast and prostate cancer cell lines demonstrated that PPARdelta expression was increased 1.5 to 3.2-fold after three hours stimulation with the natural vitamin D receptor (VDR) agonist, 1alpha,25-dihydroxyvitamin D3 (1alpha,25(OH)2D3). In silico analysis of the 20 kb of the human PPARdelta promoter revealed a DR3-type 1alpha,25(OH)2D3 response element approximately 350 bp upstream of the transcription start site, which was able to bind VDR-retinoid X receptor (RXR) heterodimers and mediate a 1alpha,25(OH)2D3-dependent upregulation of reporter gene activity. Chromatin immuno-precipitation assays demonstrated that a number of proteins representative for 1alpha,25(OH)2D3-mediated gene activation, such as VDR, RXR and RNA polymerase II, displayed a 1alpha,25(OH)2D3-dependent association with a region of the proximal PPARdelta promoter that contained the putative DR3-type VDRE. This was also true for other proteins that are involved in or are the subject of chromatin modification, such as the histone acetyltransferase CBP and histone 4, which displayed ligand-dependent association and acetylation, respectively. Finally, real-time PCR analysis demonstrated that 1alpha,25(OH)2D3 and the synthetic PPARdelta ligand L783483 show a cell and time-dependent interference in each other's effects on VDR mRNA expression, so that their combined application shows complex effects on the induction of VDR target genes, such as CYP24. Taken together, we conclude that PPARdelta is a primary 1alpha,25(OH)2D3-responding gene and that VDR and PPARdelta signaling pathways are interconnected at the level of cross-regulation of their respective transcription factor mRNA levels.
Collapse
Affiliation(s)
- Thomas W Dunlop
- Department of Biochemistry, University of Kuopio, FIN-70211 Kuopio, Finland
| | | | | | | | | | | |
Collapse
|
|
17
|
Sinkkonen L, Malinen M, Saavalainen K, Väisänen S, Carlberg C. Regulation of the human cyclin C gene via multiple vitamin D3-responsive regions in its promoter. Nucleic Acids Res 2005; 33:2440-51. [PMID: 15863722 PMCID: PMC1087898 DOI: 10.1093/nar/gki502] [Citation(s) in RCA: 57] [Impact Index Per Article: 2.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/17/2022] Open
Abstract
The candidate human tumor suppressor gene cyclin C is a primary target of the anti-proliferative hormone 1α,25-dihydroxyvitamin D3 [1α,25(OH)2D3], but binding sites for the 1α,25(OH)2D3 receptor (VDR), so-called 1α,25(OH)2D3 response elements (VDREs), have not yet been identified in the promoter of this gene. We screened various cancer cell lines by quantitative PCR and found that the 1α,25(OH)2D3 inducibility of cyclin C mRNA expression, in relationship with the 24-hydroxylase (CYP24) gene, was best in MCF-7 human breast cancer cells. To characterize the molecular mechanisms, we analyzed 8.4 kb of the cyclin C promoter by using chromatin immunoprecipitation assays (ChIP) with antibodies against acetylated histone 4, VDR and its partner receptor, retinoid X receptor (RXR). The histone 4 acetylation status of all 23 investigated regions of the cyclin C promoter did not change significantly in response to 1α,25(OH)2D3, but four independent promoter regions showed a consistent, 1α,25(OH)2D3-dependent association with VDR and RXR over a time period of 240 min. Combined in silico/in vitro screening identified in each of these promoter regions a VDRE and reporter gene assays confirmed their functionality. Moreover, re-ChIP assays monitored simultaneous association of VDR with RXR, coactivator, mediator and RNA polymerase II proteins on these regions. Since cyclin C protein is associated with those mediator complexes that display transcriptional repressive properties, this study contributes to the understanding of the downregulation of a number of secondary 1α,25(OH)2D3-responding genes.
Collapse
Affiliation(s)
| | | | | | | | - Carsten Carlberg
- To whom correspondence should be addressed. Tel: +358 17 163062; Fax: +358 17 2811510;
| |
Collapse
|
|
18
|
Väisänen S, Ryhänen S, Saarela JTA, Peräkylä M, Andersin T, Mäenpää PH. Structurally and functionally important amino acids of the agonistic conformation of the human vitamin D receptor. Mol Pharmacol 2002; 62:788-94. [PMID: 12237325 DOI: 10.1124/mol.62.4.788] [Citation(s) in RCA: 23] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022] Open
Abstract
The crystal structures of the ligand binding domain of human vitamin D receptor (VDR) complexed with its natural ligand or the superagonists MC1288 or KH1060 have recently been reported. The crystallized ligand binding domain (LBD) of VDR, however, differs from the full-length VDR with respect to deletion of 50 amino acids between its helices 2 and 3. In this study, we investigated structurally and functionally important amino acid interactions within the ligand binding pocket of the full-length VDR in the presence of several synthetic vitamin D(3) analogs. We used site-directed mutagenesis scanning combined with limited proteolytic digestion, electrophoretic mobility shift assay, and reporter gene assay and correlated the findings with the crystal structures of truncated VDR LBD. Our results suggest that structurally different agonists have distinct ligand-receptor interactions and that the amino acid residues H229, D232, E269, F279, and Y295 are critical for the agonistic conformation of the VDR. Our biological data, which were obtained with the full-length VDR, fit well with the crystal structure of the truncated VDR LBD and suggest that removal of the insertion domain between helices 2 and 3 of the receptor does not markedly influence the functionality of the VDR.
Collapse
Affiliation(s)
- Sami Väisänen
- Department of Biochemistry, University of Kuopio, Kuopio, Finland.
| | | | | | | | | | | |
Collapse
|
|
19
|
Väisänen S, Peräkylä M, Kärkkäinen JI, Steinmeyer A, Carlberg C. Critical role of helix 12 of the vitamin D(3) receptor for the partial agonism of carboxylic ester antagonists. J Mol Biol 2002; 315:229-38. [PMID: 11779241 DOI: 10.1006/jmbi.2001.5225] [Citation(s) in RCA: 47] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
Abstract
The carboxy-terminal alpha-helix of a nuclear receptor ligand-binding domain (LBD), helix 12, contains a critical, ligand-modulated interface for the interaction with coactivator proteins. In this study, using the example of the vitamin D receptor (VDR) and the partial antagonist ZK159222, the role of helix 12 (residues 417-427) for both antagonistic and agonistic receptor actions was investigated. Amino acid residue G423 was demonstrated to be critical for partial agonism of ZK159222, but not for the activity of the natural VDR agonist, 1alpha,25-dihydroxyvitamin D(3) (1alpha,25(OH)(2)D(3)). The amount of partial agonism of ZK159222 increased when helix 12 was truncated by the last four amino acid residues (Delta424-27) and augmented even more, when in addition helix 12 of VDR's dimerization partner, retinoid X receptor (RXR), was truncated. In contrast, the low agonism of a structural derivative of ZK159222, ZK168281, was not affected comparably, whereas other close structural relatives of ZK159222 even demonstrated the same agonistic activity as that of 1alpha,25(OH)(2)D(3). The amount of agonism of ZK159222 and ZK168281 at different variations of helix 12 correlated well with VDR's ability to complex with coactivator proteins and inversely correlated with the strength of the compound's antagonistic action on 1alpha,25(OH)(2)D(3) signalling. Molecular dynamics simulations of the LBD complexed with the two antagonists could explain their different action by demonstrating a more drastic displacement of helix 12 through ZK168281 than through ZK159222. Moreover, the modelling could indicate a kink of helix 12 at amino acid residue G423, which provides the last four amino acid residues of helix 12 with a modulatory role for the partial agonism of some VDR antagonists, such as ZK159222. In conclusion, partial agonism of a VDR antagonist is lower the more it disturbs helix 12 in taking the optimal position for coactivator interaction.
Collapse
MESH Headings
- Amino Acid Substitution/genetics
- Calcitriol/analogs & derivatives
- Calcitriol/pharmacology
- Computer Simulation
- Dimerization
- Electrophoretic Mobility Shift Assay
- Gene Expression Regulation/drug effects
- Genes, Reporter
- Humans
- Ligands
- Models, Molecular
- Protein Structure, Secondary
- Receptors, Calcitriol/agonists
- Receptors, Calcitriol/antagonists & inhibitors
- Receptors, Calcitriol/chemistry
- Receptors, Calcitriol/metabolism
- Receptors, Retinoic Acid/chemistry
- Receptors, Retinoic Acid/genetics
- Receptors, Retinoic Acid/metabolism
- Retinoid X Receptors
- Sequence Deletion/genetics
- Structure-Activity Relationship
- Transcription Factors/chemistry
- Transcription Factors/genetics
- Transcription Factors/metabolism
- Transfection
- Tumor Cells, Cultured
Collapse
Affiliation(s)
- Sami Väisänen
- Department of Biochemistry, University of Kuopio, Finland
| | | | | | | | | |
Collapse
|
|
20
|
Bury Y, Herdick M, Uskokovic MR, Carlberg C. Gene regulatory potential of 1alpha,25-dihydroxyvitamin D(3) analogues with two side chains. JOURNAL OF CELLULAR BIOCHEMISTRY. SUPPLEMENT 2001; Suppl 36:179-90. [PMID: 11455583 DOI: 10.1002/jcb.1082] [Citation(s) in RCA: 22] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
Abstract
The nuclear hormone 1alpha,25-dihydroxyvitamin D(3) (1alpha,25(OH)(2)D(3)) acts through the transcription factor vitamin D receptor (VDR) via combined contact with the retinoid X receptor (RXR), coactivator proteins, and specific DNA binding sites (VDREs). Ligand-mediated conformational changes of the VDR are the core of the molecular switch of nuclear 1alpha,25(OH)(2)D(3) signalling. Studying the interaction of 1alpha,25(OH)(2)D(3) analogues with this molecular switch should allow the characterization of their potential selective biological profile. A 1alpha,25(OH)(2)D(3) analogue with two side chains (Ro27-2310 or Gemini) was found to stabilize functional VDR conformations and VDR-RXR heterodimers on a VDRE with a slightly lower sensitivity than the natural hormone. A 19-nor derivative of Gemini (Ro27-5646) showed similar sensitivity whereas 5,6-trans (Ro27-6462) 3-epi (Ro27-5840) and 1alpha-fluoro (Ro27-3752) derivatives were equal to each other, but approximately 30-times less sensitive than Gemini. A des-C,D derivative of Gemini (Ro28-1909) showed only residual activity at maximal concentrations. In contrast to 1alpha,25(OH)(2)D(3), Gemini and its derivatives showed a differential preference in stabilizing VDR conformations which was found to be modulated by DNA coactivator and corepressor proteins. An analysis of the gene regulatory potential of the VDR agonists in cellular reporter gene systems demonstrated the same ranking as in the in vitro systems, but Gemini and its 19-nor derivative were found to be more sensitive than 1alpha,25(OH)(2)D(3) which indicates that the natural hormone is selectively metabolized. This study used straightforward methods for the in vitro and ex vivo evaluation of the gene regulatory potential of 1alpha,25(OH)(2)D(3) analogues. Gemini was highlighted as an interesting drug candidate which could not be optimized through obvious chemical modifications in its A-ring. J. Cell. Biochem. Suppl. 36: 179-190, 2001.
Collapse
Affiliation(s)
- Y Bury
- Institut für Physiologische Chemie I and Biomedizinisches Forschungszentrum, Heinrich-Heine-Universität Düsseldorf, Postfach 10 10 07, D-40001 Düsseldorf, Germany
| | | | | | | |
Collapse
|
|
21
|
Jensen SS, Madsen MW, Lukas J, Binderup L, Bartek J. Inhibitory effects of 1alpha,25-dihydroxyvitamin D(3) on the G(1)-S phase-controlling machinery. Mol Endocrinol 2001; 15:1370-80. [PMID: 11463860 DOI: 10.1210/mend.15.8.0673] [Citation(s) in RCA: 81] [Impact Index Per Article: 3.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/19/2022] Open
Abstract
The nuclear hormone 1alpha,25-dihydroxyvitamin D(3) induces cell cycle arrest, differentiation, or apoptosis depending on target cell type and state. Although the antiproliferative effect of 1alpha,25-dihydroxyvitamin D(3) has been known for years, the molecular basis of the cell cycle blockade by 1alpha,25-dihydroxyvitamin D(3) remains largely unknown. Here we have investigated the mechanisms underlying the G(1) arrest induced upon 1alpha,25-dihydroxyvitamin D(3) treatment of the human breast cancer cell line MCF-7. Twenty-four-hour exposure of exponentially growing MCF-7 cells to 1alpha,25-dihydroxyvitamin D(3) impeded proliferation by preventing S phase entry, an effect that correlated with appearance of the growth-suppressing, hypophosphorylated form of the retinoblastoma protein (pRb), and modulation of cyclin-dependent kinase (cdk) activities of cdk-4, -6, and -2. Time course immunochemical and biochemical analyses of the cellular and molecular effects of 1alpha,25-dihydroxyvitamin D(3) treatment for up to 6 d revealed a dynamic chain of events, preventing activation of cyclin D1/cdk4, and loss of cyclin D3, which collectively lead to repression of the E2F transcription factors and thus negatively affected cyclin A protein expression. While the observed 10-fold inhibition of cyclin D1/cdk 4-associated kinase activity appeared independent of cdk inhibitors, the activity of cdk 2 decreased about 20-fold, reflecting joint effects of the lower abundance of its cyclin partners and a significant increase of the cdk inhibitor p21(CIP1/WAF1), which blocked the remaining cyclin A(E)/cdk 2 complexes. Together with a rapid down-modulation of the c-Myc oncoprotein in response to 1alpha,25-dihydroxyvitamin D(3), these results demonstrate that 1alpha,25-dihydroxyvitamin D(3) inhibits cell proliferation by targeting several key regulators governing the G(1)/S transition.
Collapse
Affiliation(s)
- S S Jensen
- Institute of Cancer Biology, The Danish Cancer Society, DK-2100 Copenhagen, Denmark
| | | | | | | | | |
Collapse
|
|
22
|
Toell A, Gonzalez MM, Ruf D, Steinmeyer A, Ishizuka S, Carlberg C. Different Molecular Mechanisms of Vitamin D3 Receptor Antagonists. Mol Pharmacol 2001; 59:1478-85. [PMID: 11353809 DOI: 10.1124/mol.59.6.1478] [Citation(s) in RCA: 62] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022] Open
Abstract
Two structurally different antagonists of the nuclear hormone 1alpha,25-dihydroxyvitamin D(3) [1alpha,25(OH)(2)D(3)], the 25-carboxylic ester ZK159222 and the 26,23-lactone TEI-9647, have recently been described. In this study, the molecular mechanisms and the efficacy of both antagonists were compared. ZK159222 showed similar potency and sensitivity to 1alpha,25(OH)(2)D(3) in ligand-dependent gel shift assays using the vitamin D receptor (VDR), the retinoid X receptor, and specific DNA binding sites, whereas TEI-9647 displayed reduced potency and >10-fold lower sensitivity in this assay system. Limited protease digestion and gel shift clipping assays showed that the two antagonists stabilized individual patterns of VDR conformations. Both antagonists prevented the interaction of the VDR with coactivator proteins, as demonstrated by GST-pull-down and supershift assays; like the natural hormone, however, they were able to induce a dissociation of corepressor proteins. Interestingly, ZK159222 demonstrated functional antagonism in reporter gene assays both in HeLa and MCF-7 cells, whereas TEI-9647 functioned as a less sensitive antagonist only in MCF-7 cells. In conclusion, the two 1alpha,25(OH)(2)D(3) analogs act in part via different molecular mechanisms, which allows us to speculate that ZK159222 is a more complete antagonist and TEI-9647 a more selective antagonist.
Collapse
Affiliation(s)
- A Toell
- Institut für Physiologische Chemie I and Biomedizinisches Forschungszentrum, Heinrich-Heine-Universität, Düsseldorf, Germany
| | | | | | | | | | | |
Collapse
|
|
23
|
Bury Y, Ruf D, Hansen CM, Kissmeyer AM, Binderup L, Carlberg C. Molecular Evaluation of Vitamin D3 Receptor Agonists Designed for Topical Treatment of Skin Diseases11The authors declared not to have conflict of interest. J Invest Dermatol 2001; 116:785-92. [PMID: 11348471 DOI: 10.1046/j.1523-1747.2001.01332.x] [Citation(s) in RCA: 28] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/20/2022]
Abstract
MC903 (calcipotriol) is a synthetic, low calcemic analog of the nuclear hormone 1alpha,25-dihydroxyvitamin D3 and used in the treatment of psoriasis. The beneficial effects of MC903 on psoriasis are based on gene regulatory events. The genomic actions of 1alpha,25-dihydroxyvitamin D3 and its analogs are primarily mediated by a complex of the vitamin D3 receptor and the retinoid X receptor bound to a 1alpha,25-dihydroxyvitamin D3 response element that can be considered as the molecular switch of 1alpha,25-dihydroxyvitamin D3 signaling. In this study, the interaction of MC903 and two new analogs, GS1500 and EB1213, with this molecular switch was compared with that of 1alpha,25-dihydroxyvitamin D3. In DNA-dependent limited protease digestion assays, ligand-dependent gel shift assays and mammalian-one-hybrid assays, all four ligands appeared to be equally sensitive VDR agonists that activated vitamin D3 receptor-retinoid X receptor-1alpha,25-dihydroxyvitamin D3 response element complexes at a concentration of approximately 0.2 nM. The analyzed VDR agonists, however, also showed individual molecular properties, such as a reduced sensitivity in HaCaT cells (MC903), a selectivity for DNA-bound vitamin D3 receptor-retinoid X receptor heterodimers (GS1500) and a long-lasting stabilization of vitamin D3 receptor-retinoid X receptor-1alpha,25-dihydroxyvitamin D3 response element complexes (EB1213). This molecular evaluation demonstrated that the sensitivity in activating the vitamin D3 receptor is already optimal for MC903, but the analog may not be ideal in keeping the receptor active and in selectively triggering 1alpha,25-dihydroxyvitamin D3 signaling pathways.
Collapse
Affiliation(s)
- Y Bury
- Institut für Physiologische Chemie I and Biomedizinisches Forschungszentrum, Heinrich-Heine-Universität, Düsseldorf, Germany
| | | | | | | | | | | |
Collapse
|
|
24
|
Verlinden L, Verstuyf A, Quack M, Van Camp M, Van Etten E, De Clercq P, Vandewalle M, Carlberg C, Bouillon R. Interaction of two novel 14-epivitamin D3 analogs with vitamin D3 receptor-retinoid X receptor heterodimers on vitamin D3 responsive elements. J Bone Miner Res 2001; 16:625-38. [PMID: 11315990 DOI: 10.1359/jbmr.2001.16.4.625] [Citation(s) in RCA: 27] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 11/18/2022]
Abstract
This study provides a detailed and exact evaluation of the interactions between vitamin D3 receptor (VDR), retinoid X receptor (RXR), and vitamin D3 responsive elements (VDREs) mediated by two novel 14-epianalogs of 1,25-dihydroxyvitamin D [1,25(OH)2D3], 19-nor-14-epi-23-yne-1,25(OH)2D3 (TX 522) and 19-nor-14,20-bisepi-23-yne-1,25(OH)2D3 (TX 527). Both analogs were more potent (14- and 75-fold, respectively) than 1,25(OH)2D3 in inhibiting cell proliferation and inducing cell differentiation. However, DNA-independent experiments indicated that both analogs had a lower affinity to VDR and that the stability of the induced VDR conformation, as measured by limited protease digestion assays, was similar (TX 527) or even weaker (TX 522) than that induced by the parent compound. However, DNA-dependent assays such as gel shift experiments revealed that those analogs were slightly more potent (3-7 times) than 1,25(OH)2D3 in enhancing binding of VDR-RXR heterodimers to a direct repeat spaced by three nucleotides (DR3) type VDRE. The functional consequences of the ligand-VDR-RXR-VDRE interactions observed in vitro were subsequently evaluated in transfection experiments. Both 14-epianalogs enhanced transcription of VDRE containing reporter constructs more efficiently than 1,25(OH)2D3 in COS-1 and MCF-7 cells regardless of the presence of ketoconazole. Transactivation activity is suggested to be a cell-specific process because maximal transcriptional induction and the half-maximal transactivation concentration for each reporter construct were different in both cell lines. The superagonistic transactivation activity closely resembled the biological potency of these analogs on the inhibition of MCF-7 cell proliferation. These data clearly indicate that superagonistic activity starts beyond the binding of the ligand-heterodimer (VDR-RXR) complex to VDRE and thus probably involves coactivator/corepressor molecules.
Collapse
Affiliation(s)
- L Verlinden
- Laboratorium voor Experimentele Geneeskunde en Endocrinologie, Katholieke Universiteit Leuven, Belgium
| | | | | | | | | | | | | | | | | |
Collapse
|
|
25
|
Carlberg C, Quack M, Herdick M, Bury Y, Polly P, Toell A. Central role of VDR conformations for understanding selective actions of vitamin D(3) analogues. Steroids 2001; 66:213-21. [PMID: 11179728 DOI: 10.1016/s0039-128x(00)00150-1] [Citation(s) in RCA: 52] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
Abstract
The vitamin D(3) receptor (VDR) acts primarily as a heterodimer with the retinoid X receptor (RXR) on different types of 1alpha,25-dihydroxyvitamin D(3) (1alpha,25(OH)(2)D(3)) response elements (VDREs). Therefore, DNA-bound VDR-RXR heterodimers can be considered as the molecular switches of 1alpha,25(OH)(2)D(3) signalling. Functional conformations of the VDR within these molecular switches appear to be of central importance for describing the biologic actions of 1alpha,25(OH)(2)D(3) and its analogues. Moreover, VDR conformations provide a molecular basis for understanding the potential selective profile of VDR agonists, which is critical for a therapeutic application. This review discusses VDR conformations and their selective stabilization by 1alpha,25(OH)(2)D(3) and its analogues, such as EB1089 and Gemini, as a monomer in solution or as a heterodimer with RXR bound to different VDREs and complexed with coactivator or corepressor proteins.
Collapse
Affiliation(s)
- C Carlberg
- Institut für Physiologische Chemie I, Heinrich-Heine-Universität, Postfach 10 10 07, D-40001, Düsseldorf, Germany.
| | | | | | | | | | | |
Collapse
|
|
26
|
Craig TA, Benson LM, Naylor S, Kumar R. Modulation effects of zinc on the formation of vitamin D receptor and retinoid X receptor alpha-DNA transcription complexes: analysis by microelectrospray mass spectrometry. RAPID COMMUNICATIONS IN MASS SPECTROMETRY : RCM 2001; 15:1011-1016. [PMID: 11400211 DOI: 10.1002/rcm.332] [Citation(s) in RCA: 21] [Impact Index Per Article: 0.9] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/23/2023]
Abstract
The vitamin D receptor (VDR) binds zinc, and the activity of vitamin D dependent genes in cells is influenced by intracellular zinc concentrations. To determine whether zinc influences vitamin D action in cells by modulating the formation of VDR and retinoid x receptor alpha (RXR alpha) heterodimer-DNA complexes, we used microelectrospray ionization mass spectrometry (microESI-MS) to assess receptor-DNA interactions in the presence of varying amounts of zinc. In the absence of DNA, VDR and RXR alpha proteins were primarily monomeric with small amounts of protein homodimers also observed. Zn(2+) (up to 300 microM) did not change VDR or RXR alpha monomer/homodimer ratios. Mass spectra of VDR combined with RXR alpha were a sum of individual protein spectral data. Zn(2+) had no effect on the interactions of receptors. With increasing amounts of Zn(2+), additional Zn(2+) ions were detected bound to VDR and RXR alpha. microESI-MS analyses of RXR alpha in the presence of an osteopontin vitamin D DNA response element (OP-VDRE) showed RXR alpha homodimer/OP-VDRE complexes. DNA-protein complex formation increased on addition of Zn(2+) up to 200 microM; at 300 microM, Zn(2+) dissociation of the RXR alpha homodimer/OP-VDRE complexes occurred, coincident with the appearance of RXR alpha monomeric protein. When microESI-MS analyses were carried out with VDR and OP-VDRE, VDR homodimer/OP-VDRE complexes were not detected. Addition of Zn(2+) did not result in VDR/OP-VDRE complex formation. Heterodimeric VDR/RXR alpha complexes with OP-VDRE were detected by microESI-MS. Addition of 300 microM Zn(2+) resulted in dissociation of the heterodimeric VDR/RXR alpha/OP-VDRE complex. Addition of Mg(2+) in place of Zn(2+) did not alter protein/OP-VDRE complexes. Our results show that zinc modulates steroid hormone receptor-DNA interactions.
Collapse
Affiliation(s)
- T A Craig
- Nephrology Research Unit, Mayo Clinic, Rochester, MN 55905, USA
| | | | | | | |
Collapse
|
|
27
|
Herdick M, Carlberg C. Agonist-triggered modulation of the activated and silent state of the vitamin D(3) receptor by interaction with co-repressors and co-activators. J Mol Biol 2000; 304:793-801. [PMID: 11124027 DOI: 10.1006/jmbi.2000.4267] [Citation(s) in RCA: 34] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
Abstract
The nuclear receptor for the hormone 1alpha,25-dihydroxyvitamin D(3) (1alpha,25(OH)(2)D(3)), VDR, regulates gene expression via a ternary complex with the retinoid X receptor (RXR) and a 1alpha, 25(OH)(2)D(3) response element (VDRE). This complex mediates transcriptional repression through interaction with co-repressor proteins, such as NCoR, and transactivation through agonist-triggered contacts with co-activator proteins, such as SRC-1. This study demonstrates that the interaction of the VDR with NCoR results in a preferential stabilization of the VDR in a non-agonistic conformation (silent state), whereas within a complex with SRC-1 VDR is in its agonistic conformation (activated state). Helix 12 of the ligand-binding domain of the VDR was found to be a critical sensor for the differential stabilization of the activated and silent state of the receptor. VDR agonists that showed similar sensitivity in inducing VDR-RXR-VDRE complex formation were found to mediate a different dose-dependent release of NCoR from these complexes, which correlates with their ability to stabilize the silent state of the VDR in the presence of NCoR. Interestingly, up to 50 % of all VDR-NCoR complexes were found to be stable even in the presence of saturating agonist concentrations. This was confirmed by a quenching effect of overexpressed NCoR on agonist-induced gene activity mediated by VDR-RXR heterodimers. Taken together, co-activator and co-repressor proteins antagonize each other in stabilizing the activated and silent state of the receptor and modulate in this way the sensitivity and potency of the transcriptional activation by the ligand-responsive transcription factor VDR.
Collapse
Affiliation(s)
- M Herdick
- Institut für Physiologische Chemie I and Biomedizinisches Forschungszentrum, Heinrich-Heine-Universität, Düsseldorf, D-40001, Germany
| | | |
Collapse
|
|
28
|
Pérez Sestelo J, Mouriño A, Sarandeses LA. Synthesis of vitamin D(3) and calcitriol dimers as potential chemical inducers of vitamin D receptor dimerization. J Org Chem 2000; 65:8290-6. [PMID: 11101387 DOI: 10.1021/jo001084x] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/29/2022]
Abstract
The design and synthesis of vitamin D(3) dimers 2 and 3 and 1 alpha, 25-dihydroxyvitamin D(3) (calcitriol) dimers 4 and 5 are described. The dimers were designed with a view to doubly binding the vitamin D receptor (VDR) and inducing the receptor homodimerization. In the dimers the units are linked through the C-11 position in ring C by an alkyl side chain of six or 10 carbon atoms, far from the hydroxy groups responsible for the VDR binding. The linker is formed by olefin metathesis of an olefinic side chain at the C-11 position introduced by stereoselective cuprate addition. The synthesis, which is both short and convergent, uses the Wittig-Horner approach to construct the vitamin D triene system and allows the preparation of dimers with a linker of modulated length with the purpose of optimizing the vitamin D(3)-VDR interaction.
Collapse
Affiliation(s)
- J Pérez Sestelo
- Departamento de Química Fundamental, Universidade da Coruña, E-15071 A Coruña, Spain
| | | | | |
Collapse
|
|
29
|
Herdick M, Steinmeyer A, Carlberg C. Carboxylic ester antagonists of 1alpha,25-dihydroxyvitamin D(3) show cell-specific actions. CHEMISTRY & BIOLOGY 2000; 7:885-94. [PMID: 11094341 DOI: 10.1016/s1074-5521(00)00036-3] [Citation(s) in RCA: 46] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 10/18/2022]
Abstract
BACKGROUND The nuclear hormone 1alpha,25-dihydroxyvitamin D(3) (1alpha,25(OH)(2)D(3)) acts through the transcription factor vitamin D receptor (1alpha,25(OH)(2)D(3) receptor, VDR) via combined contact with the retinoid X receptor (RXR), coactivator proteins and specific DNA binding sites (1alpha,25(OH)(2)D(3) response elements, VDREs). Ligand-mediated conformational changes of the VDR are the basis of the molecular mechanisms of nuclear 1alpha,25(OH)(2)D(3) signaling. Cell-specific VDR antagonists would allow to dissect and fine regulate the pleiotropic 1alpha,25(OH)(2)D(3) endocrine system affecting the regulation of calcium homeostasis, bone mineralization and other cellular functions. RESULTS Two carboxylic ester analogues of 1alpha,25(OH)(2)D(3), ZK159222 and ZK168281, which have additional cyclopropyl rings and allylic alcohol substructures in their side chain, were characterized in different 1alpha, 25(OH)(2)D(3) target tissues as functional antagonists of 1alpha, 25(OH)(2)D(3) signaling. In all tested systems, ZK168281 showed lower residual agonistic activity and higher antagonistic effects than ZK159222, but the strength of these effects was cell-specific. Both antagonists were shown to act via the same mechanisms: they selectively stabilize an antagonistic conformation of the ligand-binding domain of the VDR within VDR-RXR-VDRE complexes, which then inhibits the interaction of the VDR with coactivator proteins and an induction of transactivation. Interestingly, cells that have been treated with antagonists were found to contain VDR-RXR heterodimers in a different conformation than cells that were stimulated with an agonist. Moreover, the strength of the functional antagonism of ZK159222 and ZK168281 appears to depend on the VDR/RXR expression ratio and high RXR levels were found to reduce the antagonistic effect of both compounds. In support of this observation, the overexpression of an transactivation function 2 (AF-2) deletion mutant of RXR resulted for both ZK159222 and ZK168281 in a reduced agonistic activity and an increased antagonistic effect. CONCLUSIONS A novel, more potent VDR antagonist, ZK168281, was identified, which stabilizes VDR-RXR heterodimers in living cells in a different conformation than agonists. In addition, the VDR/RXR ratio was found as the major discriminating factor for understanding cell-specific effects of VDR antagonists.
Collapse
MESH Headings
- Animals
- COS Cells
- Calcitriol/analogs & derivatives
- Calcitriol/chemistry
- Calcitriol/genetics
- Calcitriol/metabolism
- Calcitriol/pharmacology
- Cell Line, Transformed
- DNA/metabolism
- Dimerization
- Electrophoresis, Polyacrylamide Gel
- Female
- Genes, Reporter
- Humans
- Molecular Structure
- Rats
- Receptors, Calcitriol/antagonists & inhibitors
- Receptors, Calcitriol/chemistry
- Receptors, Calcitriol/genetics
- Receptors, Calcitriol/metabolism
- Receptors, Retinoic Acid/genetics
- Receptors, Retinoic Acid/metabolism
- Response Elements/genetics
- Retinoid X Receptors
- Transcription Factors/genetics
- Transcription Factors/metabolism
- Tumor Cells, Cultured
Collapse
Affiliation(s)
- M Herdick
- Institut für Physiologische Chemie I and Biomedizinisches Forschungszentrum, Heinrich-Heine-Universität, Düsseldorf, Germany
| | | | | |
Collapse
|
|
30
|
Bury Y, Steinmeyer A, Carlberg C. Structure activity relationship of carboxylic ester antagonists of the vitamin D(3) receptor. Mol Pharmacol 2000; 58:1067-74. [PMID: 11040055 DOI: 10.1124/mol.58.5.1067] [Citation(s) in RCA: 51] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022] Open
Abstract
A 25-carboxylic ester analog of 1alpha,25-dihydroxyvitamin D(3) [1alpha,25(OH)(2)D(3)], ZK159222 (compound 1), was recently described as a novel type of antagonist of 1alpha,25(OH)(2)D(3) signaling. In this study five derivatives of compound 1 (compounds 2-6) were selected because of their sensitivity in facilitating complex formation between the 1alpha,25(OH)(2)D(3) receptor (VDR) and the retinoid X receptor on a 1alpha,25(OH)(2)D(3) response element that was comparable to that of the natural hormone (0.2-0.9 nM). Most derivatives of compound 1 reacted as typical agonists, because they were able to promote ligand-dependent interaction of the VDR with the coactivator TIF2, stabilized the VDR preferentially in its agonistic conformation c1(LPD), and stimulated VDR-dependent gene activity with a potency similar to 1alpha,25(OH)(2)D(3). In contrast, only compound 2 showed the antagonistic profile of compound 1, which includes the incompetence to induce a VDR-TIF2 contact, the stabilization of the antagonistic conformation c2(LPD), and only a very weak and insensitive functional activity. Accordingly, only compounds 1 and 2, but not compounds 3 to 6, showed prominent antagonistic effects in cellular systems. The comparison of the structures of the compounds indicates that the essential requirements for an antagonistic function are a cyclopropyl ring at carbon 25, a hydroxy group at carbon 24, and at least a butylester. Interestingly, compound 2 was an approximately 3 times more sensitive antagonist than compound 1 and even displayed a lower residual agonistic activity. In conclusion, only a very limited number of structural variations of compound 1 are possible to keep its antagonistic profile, but the tools presented here for their in vitro evaluation allow an accurate prediction of the effects and are suited to screening for even more potent 1alpha, 25(OH)(2)D(3) antagonists.
Collapse
Affiliation(s)
- Y Bury
- Institut für Physiologische Chemie I and Biomedizinisches Forschungszentrum, Heinrich-Heine-Universität, Düsseldorf, Germany
| | | | | |
Collapse
|
|
31
|
Mahé YF, Michelet JF, Billoni N, Jarrousse F, Buan B, Commo S, Saint-Léger D, Bernard BA. Androgenetic alopecia and microinflammation. Int J Dermatol 2000; 39:576-84. [PMID: 10971723 DOI: 10.1046/j.1365-4362.2000.00612.x] [Citation(s) in RCA: 99] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 01/29/2023]
Affiliation(s)
- Y F Mahé
- Hair Biology Research Group, L'Oreal, Clichy, France
| | | | | | | | | | | | | | | |
Collapse
|
|
32
|
Polly P, Herdick M, Moehren U, Baniahmad A, Heinzel T, Carlberg C. VDR‐Alien: a novel, DNA‐selective vitamin D
3
receptor‐corepressor partnership. FASEB J 2000. [DOI: 10.1096/fasebj.14.10.1455] [Citation(s) in RCA: 28] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/09/2023]
Affiliation(s)
- Patsie Polly
- Institut für Physiologische Chemie IHeinrich‐Heine‐Universität Düsseldorf D‐40001 Germany
| | - Michaela Herdick
- Institut für Physiologische Chemie IHeinrich‐Heine‐Universität Düsseldorf D‐40001 Germany
| | - Udo Moehren
- Genetisches Institut der Justus‐Liebig‐Universität Giessen D‐35392 Germany
| | - Aria Baniahmad
- Genetisches Institut der Justus‐Liebig‐Universität Giessen D‐35392 Germany
| | | | - Carsten Carlberg
- Institut für Physiologische Chemie IHeinrich‐Heine‐Universität Düsseldorf D‐40001 Germany
| |
Collapse
|
|
33
|
Herdick M, Steinmeyer A, Carlberg C. Antagonistic action of a 25-carboxylic ester analogue of 1alpha, 25-dihydroxyvitamin D3 is mediated by a lack of ligand-induced vitamin D receptor interaction with coactivators. J Biol Chem 2000; 275:16506-12. [PMID: 10748178 DOI: 10.1074/jbc.m910000199] [Citation(s) in RCA: 60] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/06/2022] Open
Abstract
A 25-carboxylic ester analogue of 1alpha,25-dihydroxyvitamin D(3) (1alpha,25-(OH)(2)D(3)), ZK159222, was described as a novel type of antagonist of 1alpha,25-(OH)(2)D(3) signaling. The ligand sensitivity of ZK159222, in facilitating complex formation between 1alpha,25-(OH)(2)D(3) receptor (VDR) and the retinoid X receptor (RXR) on a 1alpha,25-(OH)(2)D(3) response element (VDRE), was approximately 7-fold lower when compared with 1alpha,25-(OH)(2)D(3). However, ZK159222 was not able to promote a ligand-dependent interaction of the VDR with the coactivator proteins SRC-1, TIF2, and RAC3, neither in solution nor in a complex with RXR on DNA. Functional analysis in HeLa and COS-7 cells demonstrated a 10-100-fold lower ligand sensitivity for ZK159222 than for 1alpha, 25-(OH)(2)D(3) and, most interestingly, a potency that was drastically reduced compared with 1alpha,25-(OH)(2)D(3). A cotreatment of 1alpha,25-(OH)(2)D(3) with a 100-fold higher concentration of ZK159222 resulted in a prominent antagonistic effect both in functional in vivo and in in vitro assays. These data suggest that the antagonistic action of ZK159222 is due to a lack of ligand-induced interaction of the VDR with coactivators with a parallel ligand sensitivity, which is sufficient for competition with the natural hormone for VDR binding.
Collapse
Affiliation(s)
- M Herdick
- Institut für Physiologische Chemie I and Biomedizinisches Forschungszentrum, Heinrich-Heine-Universität, D-40001 Düsseldorf, Germany
| | | | | |
Collapse
|
|
34
|
Kröncke KD, Carlberg C. Inactivation of zinc finger transcription factors provides a mechanism for a gene regulatory role of nitric oxide. FASEB J 2000; 14:166-73. [PMID: 10627291 DOI: 10.1096/fasebj.14.1.166] [Citation(s) in RCA: 91] [Impact Index Per Article: 3.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/11/2022]
Abstract
Nitric oxide (NO) is known to induce Zn(2+) release from the zinc-storing protein metallothionein and to induce Zn(2+) release within the nuclei and cytoplasm of cells. This suggests that zinc finger proteins may be primary targets of NO-induced stress. In this study, the specific interaction of the heterodimeric complex of two zinc finger transcription factors, 1alpha,25-dihydroxyvitamin D(3) (1alpha,25(OH)(2)D(3)) receptor (VDR) and retinoid X receptor (RXR) with 1alpha,25(OH)(2)D(3) response elements (VDREs), was used as a model system. NO was applied to this system via the NO donors SNOC and MAMA/NO and caused a dose-dependent inhibition of VDR-RXR-VDRE complex formation (IC(50) values 0.5-0.8 mM). Ligand-bound or preformed complexes displayed less sensitivity to NO-induced stress. These in vitro effects of NO were found to be reversible. Functional assays in transiently transfected cells indicated that NO can also act in vivo as a repressor of 1alpha,25(OH)(2)D(3) signaling (IC(50) value of the slow NO donor DETA/NO, 0.5 mM). These findings suggest that NO has a modulatory role on transcription factors depending on their sensitivity to NO-induced stress, thus providing a mechanism for a gene regulatory function of NO.
Collapse
Affiliation(s)
- K D Kröncke
- Research Group Immunobiology, Institut für Physiologische Chemie I, Heinrich-Heine-Universität Düsseldorf, Germany.
| | | |
Collapse
|
|
35
|
Craig TA, Benson LM, Tomlinson AJ, Veenstra TD, Naylor S, Kumar R. Analysis of transcription complexes and effects of ligands by microelectrospray ionization mass spectrometry. Nat Biotechnol 1999; 17:1214-8. [PMID: 10585721 DOI: 10.1038/70767] [Citation(s) in RCA: 33] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/09/2022]
Abstract
The human vitamin D receptor (VDR) and retinoid X receptor-alpha (RXRalpha) modulate gene activity by forming homodimeric or heterodimeric complexes with specific DNA sequences and interaction with other elements of the transcriptional apparatus in the presence of their known endogenous ligands 1alpha,25-dihydroxyvitamin D3 (1, 25-[OH]2D3) and 9-cis-retinoic acid (9-c-RA). We used rapid buffer exchange gel filtration in conjunction with microelectrospray ionization mass spectrometry (microESI-MS) to study the binding of these receptors to the osteopontin vitamin D response element (OP VDRE). In the absence of DNA, both VDR and RXRalpha existed primarily as monomers, but in the presence of OP VDRE, homodimeric RXRalpha and heterodimeric RXRalpha-VDR complexes were shown to bind OP VDRE. Addition of 9-c-RA increased RXRalpha homodimer-OP VDRE complexes, and addition of 1,25-(OH) 2D3 resulted in formation of 1, 25-(OH)2D 3-VDR-RXRalpha-OP VDRE complexes. Addition of low-affinity binding ligands had no detectable effect on the VDR-RXRalpha-OP VDRE transcription complex. These results demonstrate the utility of microESI-MS in analyzing multimeric, high-molecular-weight protein-protein and protein-DNA complexes, and the effects of ligands on these transcriptional complexes.
Collapse
Affiliation(s)
- T A Craig
- Nephrology Research Unit, Mayo Clinic, Rochester, MN 55905, USA
| | | | | | | | | | | |
Collapse
|
|
36
|
Toell A, Degenhardt S, Grabensee B, Carlberg C. Inhibitory effect of uremic solutions on protein-DNA-complex formation of the vitamin D receptor and other members of the nuclear receptor superfamily. J Cell Biochem 1999. [DOI: 10.1002/(sici)1097-4644(19990901)74:3<386::aid-jcb7>3.0.co;2-1] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/06/2022]
|
|
37
|
Reinhardt TA, Koszewski NJ, Omdahl J, Horst RL. 1,25-Dihydroxyvitamin D(3) and 9-cis-retinoids are synergistic regulators of 24-hydroxylase activity in the rat and 1, 25-dihydroxyvitamin D(3) alters retinoic acid metabolism in vivo. Arch Biochem Biophys 1999; 368:244-8. [PMID: 10441374 DOI: 10.1006/abbi.1999.1335] [Citation(s) in RCA: 10] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
Abstract
The RXR forms a heterodimer with the VDR to activate genes that are regulated by 1,25(OH)(2)D(3). In the absence of RXR's ligand, 9-cis-RA, RXR appears to be a silent partner to VDR. The effect of 9-cis-RA on VDR/RXR heterodimer formation and 1, 25(OH)(2)D(3)-mediated gene expression in vivo remains unclear. We examined the effect of exogenous 9-cis-RA or 9-cis-RA precursors, 9, 13-di-cis-RA and 9-cis-RCHO, on 1,25(OH)(2)D(3)-mediated induction rat renal 24-hydroxylase. The rats were treated as follows: (1) vehicle; (2) 1,25(OH)(2)D(3); (3) 1,25(OH)(2)D(3) + 9-cis-RA; (4) 1, 25(OH)(2)D(3) + 9,13-di-cis-RA; (5) 1,25(OH)(2)D(3) + 9-cis-RCHO; (6) 9-cis-RA; (7) 9,13-di-cis-RA; and (8) 9-cis-RCHO. 1, 25(OH)(2)D(3) was administered IP 18 h prior to sacrifice. The retinoids were administered every 4 h, starting 28 h prior to sacrifice. The last retinoid dose was administered 4 h prior to sacrifice. Treatment with 1,25(OH)(2)D(3) alone increased 24-hydroxylase from 35 +/- 6 (controls) to 258 +/- 44 pmol/min/g tissue. When 1,25(OH)(2)D(3) was administered with 9-cis-RA, 9, 13-di-cis-RA, or 9-cis-RCHO, 24-hydroxylases were 568 +/- 56, 524 +/- 56, and 463 +/- 62 pmol/min/g tissue, respectively. Furthermore, codosing of 1,25(OH)(2)D(3) and 9-cis-retinoids resulted in higher circulating concentrations of 9-cis-RA and 9,13-di-cis-RA when compared to rats dosed with 9-cis-retinoids alone. This was shown to be due to 1,25(OH)(2)D(3) increasing the half-life of 9,13-di-cis-RA by three to four times. These results show that 9-cis-RA can act synergistically with 1,25(OH)(2)D(3) in the regulation of 24-hydroxylase in vivo. Additionally, 1,25(OH)(2)D(3) regulates 9, 13-di-cis-RA metabolism in vivo.
Collapse
Affiliation(s)
- T A Reinhardt
- Agricultural Research Service, United States Department of Agriculture, Ames, Iowa, 50010, USA.
| | | | | | | |
Collapse
|
|
38
|
Craig TA, Lutz WH, Kumar R. Association of prokaryotic and eukaryotic chaperone proteins with the human 1alpha,25-dihydroxyvitamin D(3) receptor. Biochem Biophys Res Commun 1999; 260:446-52. [PMID: 10403788 DOI: 10.1006/bbrc.1999.0931] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/22/2022]
Abstract
Steroid hormone receptors (SHR) form complexes with heat shock proteins (hsps). The 1alpha,25-dihydroxyvitamin D(3) receptor (VDR) has not been previously shown to interact with hsps. During expression and purification of VDR-glutathione S-transferase (VDR-GST) fusion proteins encompassing full-length, DNA, and ligand-binding domains of the VDR (FL-VDR, DBD-VDR, and LBD-VDR), we observed binding of bacterial hsps with VDR-GST constructs. All VDR constructs bound DnaK in amounts greater than GST alone and bound smaller amounts of DnaJ or GrpE. GroEL bound only to FL-VDR. GroES did not bind to VDR. When VDR-GST constructs were incubated with a reticulocyte lysate system that has been used previously to examine SHR-hsp interactions, eukaryotic hsc70 was detected bound to FL-VDR and DBD-VDR. Binding of hsp90 to VDR was not detected. However, geldanamycin, an hsp90 inhibitor, reduced 1alpha,25-dihydroxyvitamin D(3)-mediated gene activation in osteoblasts. Our data show that the bacterial and eukaryotic hsps associate with the VDR and might be involved in VDR function.
Collapse
Affiliation(s)
- T A Craig
- Department of Medicine, Mayo Clinic/Foundation, Rochester, Minnesota, 55905, USA
| | | | | |
Collapse
|
|
39
|
Defacque H, Piquemal D, Basset A, Marti J, Commes T. Transforming growth factor-beta1 is an autocrine mediator of U937 cell growth arrest and differentiation induced by vitamin D3 and retinoids. J Cell Physiol 1999; 178:109-19. [PMID: 9886497 DOI: 10.1002/(sici)1097-4652(199901)178:1<109::aid-jcp14>3.0.co;2-x] [Citation(s) in RCA: 43] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/26/2022]
Abstract
Vitamin D and retinoids cooperate to inhibit the proliferation and induce the differentiation of human myelomonocytic U937 leukemia cells. In the present work, we investigated the role of TGF-beta as an endogenous mediator of this process. We found that the TGF-beta1 precursor began to accumulate in cell culture supernatants soon after the addition of 1alpha,25 dihydroxyvitamin D3 (VD) and retinoids. We used neutralizing antibodies (AbTGF-beta) and antisense oligonucleotide (AS Oligo) to inhibit its possible effects. Our data demonstrated that AbTGF-beta partially inhibit the expression of the differentiated phenotype, as assessed by measurement of phagocytic activity, response to the chemotactic peptide fMLP, and lysozyme secretion. AS Oligo was also inhibitory, and the effects of AS Oligo and AbTGF-beta were cumulative. Cell growth inhibition induced by VD and retinoids was completely reversed, and differentiation was reduced by about 75% when both inhibitors were associated. Time course experiments based on the delayed addition of AbTGF-beta and AS Oligo showed that TGF-beta1 was required for cell differentiation 24 h after the addition of inducers. Studies on TGF-beta receptors revealed that, while the expression of type II receptor was stable, the level of type I TGF-beta receptor mRNA and the expression of the protein began to decline early during the differentiation process. As a whole, these results support the notion that an autocrine TGF-beta pathway, activated by VD and retinoids in U937 cells, is involved in the early steps of the process leading to cell growth arrest and differentiation.
Collapse
Affiliation(s)
- H Defacque
- EMBL, Department of Cell Biology, Heidelberg, Germany
| | | | | | | | | |
Collapse
|
|
40
|
Quack M, Clarin A, Binderup E, Björkling F, Hansen CM, Carlberg C. Structural variants of the vitamin D analogue EB1089 reduce its ligand sensitivity and promoter selectivity. J Cell Biochem 1998. [DOI: 10.1002/(sici)1097-4644(19981201)71:3<340::aid-jcb3>3.0.co;2-c] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/07/2022]
|
|
41
|
Quack M, Szafranski K, Rouvinen J, Carlberg C. The role of the T-box for the function of the vitamin D receptor on different types of response elements. Nucleic Acids Res 1998; 26:5372-8. [PMID: 9826761 PMCID: PMC148003 DOI: 10.1093/nar/26.23.5372] [Citation(s) in RCA: 26] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/12/2022] Open
Abstract
The nuclear hormone 1alpha,25-dihydroxyvitamin D3(VD) mainly functions through a heterodimer formed between the VD receptor (VDR) and the retinoid X receptor (RXR). This transcription factor complex specifically recognizes DNA sequences, referred to as VD response elements (VDREs), that are formed by two hexameric core binding motifs arranged either as direct repeats spaced by 3 nt (DR3) or inverted palindromes with nine intervening nucleotides (IP9). Gel shift clipping assays provided the first evidence that VDR-RXR heterodimers form different conformations on these two types of VDREs. Since the T-box within the C-terminal extension of the receptor DNA binding domain (DBD) was previously shown to form a dimerization interface with the partner receptor DBD when bound to DR-type response elements, all six amino acid residues of the VDR T-box were investigated for their role in VDR-RXR heterodimer complex formation on DR3- and IP9-type VDREs. Interestingly, the residue Phe93 (F93) was found to be critical on both types of VDREs, whereas the role of the residue Ile94 (I94) was found to depend on ionic strength of the binding reaction and the nature of the VDRE. However, under physiological conditions I94 was also shown to be critical on both VDRE types. The monitored differences between the two VDR-containing protein-DNA complexes helps in an understanding of the differential action of the nuclear hormone VD and its therapeutically important analogues.
Collapse
Affiliation(s)
- M Quack
- Institut für Physiologische Chemie I and Biomedizinisches Forschungszentrum, Heinrich-Heine-Universität,D-40001 Düsseldorf, Germany
| | | | | | | |
Collapse
|
|
42
|
Quack M, Mørk Hansen C, Binderup E, Kissmeyer AM, Carlberg C. Metabolism of the vitamin D3 analogue EB1089 alters receptor complex formation and reduces promoter selectivity. Br J Pharmacol 1998; 125:607-14. [PMID: 9831892 PMCID: PMC1571001 DOI: 10.1038/sj.bjp.0702086] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022] Open
Abstract
1. 1alpha,25-dihydroxyvitamin3 (VD) is a nuclear hormone that has important cell regulatory functions but also a strong calcemic effect. EB1089 is a potent antiproliferative VD analogue, which has a modified side chain resulting in increased metabolic stability and a selective functional profile. Since EB1089 is considered for potential systemic application, it will be investigated to what extent its recently identified metabolites (hydroxylated at positions C26 and C26a) contribute to biological profile of the VD analogue. 2. Limited protease digestion analysis demonstrated that EB1089 is able to stabilize the high affinity ligand binding conformation of the VDR, starting at concentrations of 0.1 nM and affecting up to 80% of all receptor molecules. The metabolites EB1445 and EB1470 showed to be 100 fold less potent than EB1089, whereas the remaining three metabolites (EB1435, EB1436 and EB1446) showed a clearly reduced ability to stabilize the high affinity ligand binding conformation. Interestingly, at pharmacological concentrations all EB1089 metabolites stabilized a second, apparently lower affinity conformation to a much higher extent than EB1089. 3. In reporter gene assays all metabolites showed lower potency than EB1089. Moreover, the preference of EB1089 for activation of VDR binding to sites formed by inverted palindromic arrangements spaced by nine nucleotide (IP9-type VD response elements) appeared to be reduced (with EB1445 and EB1470) or completely lost (with EB1435, EB1436 and EB1446). The ranking of EB1089 and its metabolites that was obtained by limited protease digestion and reporter gene assays was confirmed by an analysis of their antiproliferative effect in breast cancer cells. . The potency and selectivity of the EB1089 metabolites in mediating gene regulatory effects was found to be drastically reduced in comparison to the parent compound suggesting that the contribution of the metabolites to the biological effect of EB1089 is minor. However, the compounds showed to be interesting tools for understanding the selective biological profile of EB1089.
Collapse
Affiliation(s)
- M Quack
- Institut für Physiologische Chemie I, Heinrich-Heine-Universität, Düsseldorf, Germany
| | | | | | | | | |
Collapse
|
|
43
|
Dong D, Noy N. Heterodimer formation by retinoid X receptor: regulation by ligands and by the receptor's self-association properties. Biochemistry 1998; 37:10691-700. [PMID: 9692959 DOI: 10.1021/bi980561r] [Citation(s) in RCA: 42] [Impact Index Per Article: 1.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/08/2023]
Abstract
The retinoid X receptor (RXR), a nuclear receptor that is activated by 9-cis-retinoic acid (9cRA), can regulate transcription as a homodimer or as a heterodimer with numerous other receptors. It was previously shown that, in the absence of ligand, RXR self-associates into homotetramers which are transcriptionally silent, and that ligand-binding induces dissociation of RXR tetramers into active species, dimers and monomers. Here, the implications of tetramer formation by RXR for the ability of the receptor to heterodimerize with the retinoic acid and the vitamin D receptors (RAR and VDR) were studied. In addition, the effects of cognate ligands for RXR and for RAR and VDR on formation of the respective heterodimers were examined. The data indicate that RXR subunits that are sequestered in tetramers were not available for interactions with RAR or VDR and, consequently, that in the absence of a RXR ligand, only a small fraction of this receptor became involved in heterodimers. RXR-selective ligands led to tetramer dissociation, but also inhibited the formation of heterodimers, directing a significant fraction of RXR into homodimers. Ligand binding by either heterodimerization partner significantly stabilized the respective heterodimer. Thus, maximal heterodimerization was observed in the presence of both 9cRA, acting to release active RXR species from tetramers, and the partner's cognate ligand, acting to overcome the inhibitory effect of 9cRA on heterodimer formation. These observations suggest that, by modulating protein-protein interactions within homo- and hetero-oligomers of RXR, cognate ligands control the relative distribution of potential RXR-containing complexes, thereby determining the transcriptional pathways that may be invoked under particular conditions in vivo.
Collapse
Affiliation(s)
- D Dong
- Division of Nutritional Science, Cornell University, Ithaca, New York 14853, USA
| | | |
Collapse
|
|
44
|
van de Kerkhof PC, Cambazard F, Hutchinson PE, Haneke E, Wong E, Souteyrand P, Damstra RJ, Combemale P, Neumann MH, Chalmers RJ, Olsen L, Revuz J. The effect of addition of calcipotriol ointment (50 micrograms/g) to acitretin therapy in psoriasis. Br J Dermatol 1998; 138:84-9. [PMID: 9536227 DOI: 10.1046/j.1365-2133.1998.02030.x] [Citation(s) in RCA: 74] [Impact Index Per Article: 2.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/07/2023]
Abstract
Our purpose was to find out whether the addition of calcipotriol ointment (50 micrograms/g) to systemic treatment with acitretin produces additional therapeutic effects and thereby an acitretin-sparing effect, and further to investigate the safety and tolerability of this combination. A multicentre, randomized, double-blind placebo-controlled study was designed. Patients were randomized to receive calcipotriol or placebo. All patients were treated with a starting dose of 20 mg acitretin per day and doses were adjusted at 2-weekly intervals with increments of 10 mg per day up to a maximum of 70 mg per day. The dose requirement for acitretin, clinical signs and adverse events were recorded. Seventy-six patients were randomized to treatment with calcipotriol 50 micrograms/g ointment twice daily and 59 patients to treatment with the vehicle only twice daily. Clearance or marked improvement was achieved by 67% of the patients in the calcipotriol group and by 41% of the patients in the placebo group (P = 0.006). Calcipotriol treatment proved to have a statistically significant additional effect to acitretin on the Psoriasis Area and Severity Index, redness, thickness and scaliness as compared with placebo. Clearance or marked improvement was achieved with a statistically significantly lower cumulative dose of acitretin by the patients in the calcipotriol group as compared with the placebo group. The number of patients reporting adverse events was pronounced and largely related to acitretin. No significant differences were observed between the two treatment groups with respect to adverse events. Laboratory assessments were essentially normal. The addition of calcipotriol ointment to acitretin treatment contributes to the efficacy, reduces the cumulative dose of acitretin to reach marked improvement or clearance, and is well-tolerated and safe.
Collapse
Affiliation(s)
- P C van de Kerkhof
- Department of Dermatology, University Hospital Nijmegen, The Netherlands
| | | | | | | | | | | | | | | | | | | | | | | |
Collapse
|
|
45
|
Wirtanen L, Huard V, Séguin C. Molecular cloning from neurulating Ambystoma mexicanum embryos of the cDNA encoding an orphan nuclear receptor (aDOR1) closely related to TR2-11. Differentiation 1997; 62:159-70. [PMID: 9503600 DOI: 10.1046/j.1432-0436.1998.6240159.x] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/06/2023]
Abstract
We have isolated a cDNA encoding a novel orphan nuclear receptor, aDOR1, closely related to testicular receptor-2 (TR2) orphan receptor family members, from neurulating Ambystoma mexicanum embryos. The cDNA sequence predicts a protein primary sequence of 416 amino acids with a calculated molecular weight of 45.8 kDa. While the DNA-binding domains of aDOR1 and hTR2-11 share 96% identity, considerable divergence is observed at both extremities of the peptides. At the N-terminus, aDOR1 is 66% identical to hTR2-11 and longer by 37 amino acids. At the C-terminus, despite a greater similarity (69%), aDOR1 is much shorter than the hTR2 isoforms and seems to encode a distinct ligand-binding domain. Expression of aDOR1 was studied by the reverse transcription polymerase chain reaction assay (RT-PCR). High mRNA levels were detected during oogenesis, they remained high during the cleavage stage, and decreased at the mid-blastula transition (MBT). Transcripts increased again at the end of gastrulation, reached a peak level during neurulation, and leveled off after closure of the neural tube. In neurulas dissected along the anteroposterior axis, aDOR1 mRNA was enriched at both extremities of the embryo, while no particular distribution was favored along the dorsoventral axis. Retinoic acid (RA) treatments at the beginning of gastrulation did not affect overall mRNA levels in the neurula nor its distribution along both axes. In the adult, expression was predominant in the brain; lower levels (about 15%) were detectable in all germ layer derivatives, except muscle. These results suggest that aDOR1 may be required for the early determination events occurring during the cleavage stages of development, and may be involved in embryogenesis and in brain function.
Collapse
MESH Headings
- Ambystoma/embryology
- Amino Acid Sequence
- Animals
- Base Sequence
- Blastocyst
- Blotting, Southern
- Brain/growth & development
- Brain/metabolism
- Cloning, Molecular
- DNA-Binding Proteins
- Embryo, Nonmammalian/drug effects
- Embryo, Nonmammalian/physiology
- Gene Expression Regulation, Developmental
- Humans
- Mice
- Molecular Sequence Data
- Nervous System/embryology
- Nuclear Receptor Subfamily 2, Group C, Member 1
- Receptors, Cytoplasmic and Nuclear/genetics
- Receptors, Cytoplasmic and Nuclear/metabolism
- Receptors, Thyroid Hormone/genetics
- Sequence Analysis
- Tissue Distribution
- Transcription, Genetic
- Tretinoin/pharmacology
Collapse
Affiliation(s)
- L Wirtanen
- Centre de Recherche en Cancérologie de l'Université Laval, Centre Hospitalier Universitaire de Québec, Canada
| | | | | |
Collapse
|
|
46
|
Polly P, Carlberg C, Eisman JA, Morrison NA. 1 alpha,25-dihydroxyvitamin D3 receptor as a mediator of transrepression of retinoid signaling. J Cell Biochem 1997; 67:287-96. [PMID: 9361184 DOI: 10.1002/(sici)1097-4644(19971201)67:3<287::aid-jcb1>3.0.co;2-s] [Citation(s) in RCA: 16] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/05/2023]
Abstract
The receptors for retinoic acid (RA) and for 1 alpha,25-dihydroxyvitamin D3 (VD), RAR, RXR, and VDR are ligand-inducible members of the nuclear receptor superfamily. These receptors mediate their regulatory effects by binding as dimeric complexes to response elements located in regulatory regions of hormone target genes. Sequence scanning of the tumor necrosis factor-alpha type 1 receptor (TNF alpha RI) gene identified a 3' enhancer region composed of two directly repeated hexameric core motifs spaced by 2 nucleotides (DR2). On this novel DR2-type sequence, but not on a DR5-type RA response element, VD was shown to act through its receptor, the vitamin D receptor (VDR), as a repressor of retinoid signalling. The repression appears to be mediated by competitive protein-protein interactions between VDR, RAR, RXR, and possibly their cofactors. This VDR-mediated transrepression of retinoid signaling suggests a novel mechanism for the complex regulatory interaction between retinoids and VD.
Collapse
MESH Headings
- 3T3 Cells
- Animals
- Antigens, CD/genetics
- Binding, Competitive
- Calcitriol/metabolism
- Cholecalciferol
- DNA, Recombinant
- Humans
- Mice
- Osteocalcin/genetics
- Promoter Regions, Genetic/genetics
- Receptors, Calcitriol/genetics
- Receptors, Retinoic Acid/genetics
- Receptors, Tumor Necrosis Factor/genetics
- Receptors, Tumor Necrosis Factor, Type I
- Retinoic Acid Receptor alpha
- Retinoid X Receptors
- Signal Transduction/genetics
- Transcription Factors/genetics
- Transcription, Genetic/genetics
- Tretinoin/metabolism
Collapse
Affiliation(s)
- P Polly
- Bone and Mineral Program, Garvan Institute of Medical Research, Sydney, Australia
| | | | | | | |
Collapse
|
|
47
|
Kahlen JP, Carlberg C. Allosteric interaction of the 1alpha,25-dihydroxyvitamin D3 receptor and the retinoid X receptor on DNA. Nucleic Acids Res 1997; 25:4307-13. [PMID: 9336462 PMCID: PMC147055 DOI: 10.1093/nar/25.21.4307] [Citation(s) in RCA: 15] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/05/2023] Open
Abstract
Genomic actions of the hormone 1alpha,25-dihydroxy-vitamin D3(VD) are mediated by the transcription factor VDR, which is a member of the nuclear receptor superfamily. VDR acts in most cases as a heterodimeric complex with the retinoid X receptor (RXR) from specific DNA sequences in the promoter of VD target genes called VD response elements (VDREs). This study describes a mutation (K45A) of the VDR DNA binding domain that enhances the affinity and ligand responsiveness of VDR-RXR heterodimers on some VDREs. In analogy to a homologous mutation in the glucocorticoid receptor (K461A), this lysine residue appears to function as an allosteric 'lock'. Interestingly, overexpression of RXR was found to reduce the responsiveness and sensitivity of wild type VDR to VD, but enhance the response of VDRK45A. Moreover, the transactivation domains of both VDR and RXR were shown to be essential for obtaining responsiveness of the heterodimers to VD and 9- cis retinoic acid (the RXR ligand). This indicates that RXR is an active rather than silent partner of the VDR on the VDREs tested. Taken together, transactivation by VDR-RXR heterodimers can be triggered individually by all components of the protein-DNA complex, but full potency appears to be reached through allosteric interaction.
Collapse
Affiliation(s)
- J P Kahlen
- Clinique de Dermatologie, Hôpital Cantonal Universitaire, CH-1211 Genève 14, Switzerland and Institut für Physiologische Chemie I, Heinrich-Heine-Universität Düsseldorf, D-40001 Düsseldorf, Germany
| | | |
Collapse
|
|
48
|
Danielsson C, Mathiasen IS, James SY, Nayeri S, Bretting C, Hansen CM, Colston KW, Carlberg C. Sensitive induction of apoptosis in breast cancer cells by a novel 1,25-dihydroxyvitamin D3 analogue shows relation to promoter selectivity. J Cell Biochem 1997; 66:552-62. [PMID: 9282333 DOI: 10.1002/(sici)1097-4644(19970915)66:4<552::aid-jcb14>3.0.co;2-d] [Citation(s) in RCA: 70] [Impact Index Per Article: 2.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/05/2023]
Abstract
The biologically active form of vitamin D3, the nuclear hormone 1 alpha,25-dihydroxyvitamin D3 (VD), is an important regulator of cellular growth, differentiation, and death. The hormone mediates its action through the activation of the transcription factor VDR, which is a member of the superfamily of nuclear receptors. In most cases the ligand-activated VDR is found in complex with the retinoid X receptor (RXR) and stimulates gene transcription mainly from VD response elements (VDREs) that are formed by two hexameric core binding motifs and are arranged either as a direct repeat spaced by three nucleotides (DR3) or as an inverted palindrome spaced by nine nucleotides (1P9). The two VD analogues CB1093 and EB1089 are both very potent inhibitors of the proliferation of MCF-7 cultured breast cancer cells displaying approximately 100-fold lower IC50 values (0.1 nM) than the natural hormone. In addition, CB1093 is even more potent in vivo than EB1089 in producing regression of experimental mammary tumors. Moreover, both VD analogues induce apoptosis in MCF-7 cells, but CB1093 is effective at concentrations approximately 10-fold lower than EB1089. In accordance, the reduction of Bcl-2 protein expression showed CB1093 to be more potent than EB1089. This suggests that the antiproliferative effect of CB1093 may be related mainly to its apoptosis inducing effect, whereas EB1089 may preferentially have effects on growth arrest. EB1089 is known to result in a selectivity for the activation of IP9-type VDREs, whereas CB1093 shows a preference for the activation of DR3-type VDREs. This promoter selectivity suggests that the effects of VD and its analogues on growth arrest and the induction of apoptosis may be mediated by different primary VD responding genes. In conclusion, CB1093 was found to be a potent inhibitor of rat mammary tumor growth in vivo. CB1093 also displayed a high potency in vitro in the induction of apoptosis, a process that may be linked to a promoter selectivity for DR3-type VDREs.
Collapse
MESH Headings
- Animals
- Apoptosis/drug effects
- Apoptosis/genetics
- Apoptosis/physiology
- Breast Neoplasms/pathology
- Calcitriol/administration & dosage
- Calcitriol/analogs & derivatives
- Calcitriol/chemistry
- Calcitriol/pharmacology
- Calcium/metabolism
- Cell Division/drug effects
- Female
- Gene Expression Regulation, Neoplastic
- Humans
- Promoter Regions, Genetic/drug effects
- Promoter Regions, Genetic/genetics
- Promoter Regions, Genetic/physiology
- Proto-Oncogene Proteins c-bcl-2/analysis
- Proto-Oncogene Proteins c-bcl-2/drug effects
- Rats
- Rats, Inbred Lew
- Rats, Inbred Strains
- Rats, Wistar
- Receptors, Calcitriol/physiology
- Transcription, Genetic/genetics
- Transcription, Genetic/physiology
- Tumor Cells, Cultured
Collapse
Affiliation(s)
- C Danielsson
- Clinique de Dermatologie, Hôpital Cantonal Universitaire, Genève, Switzerland
| | | | | | | | | | | | | | | |
Collapse
|
|
49
|
Craig TA, Veenstra TD, Naylor S, Tomlinson AJ, Johnson KL, Macura S, Juranić N, Kumar R. Zinc binding properties of the DNA binding domain of the 1,25-dihydroxyvitamin D3 receptor. Biochemistry 1997; 36:10482-91. [PMID: 9265628 DOI: 10.1021/bi970561b] [Citation(s) in RCA: 32] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/05/2023]
Abstract
To assess the zinc binding stoichiometry and the structural changes induced upon the binding of zinc to the human vitamin D receptor (VDR), we expressed the DNA binding domain (DBD) of the human VDR in bacteria as a soluble glutathione-S-transferase fusion protein at 20 degrees C, and examined the apo-protein and metal-liganded protein by mass spectrometry, and circular dichroism and nuclear magnetic resonance spectroscopy. Following final preparation with a zinc-free buffer, the VDR DBD bound 2 mol of zinc/mol of protein as measured by inductively coupled plasma-mass spectrometry and electrospray ionization-mass spectrometry. When protein preparation was carried out in a zinc containing buffer and zinc content of the protein was assesed by the same methods, VDR DBD bound 4 mol of zinc/mol of protein. Analysis of the protein using circular dichroism spectroscopy demonstrated that the EDTA-treated protein increased in alpha-helical content from 16 to 27% on the addition of zinc. Equilibrium ultracentrifugal analyses of the VDR DBD indicated that the protein was present in solution as a monomer. Gel mobility shift analyses of the VDR DBD with several vitamin D response elements (VDREs) in the absence of accessory proteins such as retinoic acid receptor, showed that VDR DBD was able to form a protein/VDRE DNA structural complex. In the presence of zinc, proton NMR NOESY spectra showed that the protein possessed elements of secondary structure. The addition of VDRE DNA, but not random DNA, caused changes in the proton NMR spectra of VDRE DNA indicating specific interaction between protein and DNA groups. We conclude that the DBD of the VDR binds zinc and DNA and undergoes conformational changes on binding to the metal and DNA.
Collapse
Affiliation(s)
- T A Craig
- Department of Medicine, Mayo Clinic and Foundation, Rochester, Minnesota 55905, USA
| | | | | | | | | | | | | | | |
Collapse
|
|
50
|
Schräder M, Kahlen JP, Carlberg C. Functional characterization of a novel type of 1 alpha,25-dihydroxyvitamin D3 response element identified in the mouse c-fos promoter. Biochem Biophys Res Commun 1997; 230:646-51. [PMID: 9015378 DOI: 10.1006/bbrc.1996.6025] [Citation(s) in RCA: 34] [Impact Index Per Article: 1.2] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 02/03/2023]
Abstract
The seco-steroid 1 alpha,25-dihydroxyvitamin D3 (VD) is known to inhibit cellular proliferation and to induce differentiation as well as programmed cell death (apoptosis). VD is the ligand of the transcription factor VDR, which is a member of the nuclear receptor superfamily. Primary VD responding genes contain a VD response element (VDRE), on which VDR binds as a dimeric complex. The main heterodimeric partner of VDR is the retinoid X receptor (RXR) and the majority of the known natural VDREs are formed by a direct repeat of hexameric core binding motifs spaced by 3 nucleotides. Most of the genes carrying DR3-type VDREs are associated with the hormone's classical function, which is the regulation of calcium homeostasis. Recently, it has been found that inverted palindromic arrangements spaced by 9 nucleotides also form functional VDREs. This paper reports the identification of a novel IP9-type VDRE in the mouse c-fos promoter. This elements is bound with high affinity by VDR-RXR heterodimers and responds at 10-fold lower concentrations to the potent anti-proliferative VD analogue EB1089 than to VD. This suggests that VD may be directly involved in the transcriptional regulation of the cell cycle via the activation of the c-fos gene.
Collapse
Affiliation(s)
- M Schräder
- Clinique de Dermatologie, Hôpital Cantonal Universitaire, Genève, Switzerland
| | | | | |
Collapse
|