Please wait a minute...
 Home  About the Journal Editorial Board Aims & Scope Peer Review Policy Subscription Contact us
 
Early Edition  //  Current Issue  //  Open Special Issues  //  Archives  //  Most Read  //  Most Downloaded  //  Most Cited
Aging and disease    2020, Vol. 11 Issue (2) : 241-253     DOI: 10.14336/AD.2019.0618
Orginal Article |
Divergent Effect of Tacalcitol (PRI-2191) on Th17 Cells in 4T1 Tumor Bearing Young and Old Ovariectomized Mice
Agata Pawlik1, Artur Anisiewicz1, Beata Filip-Psurska1, Dagmara Klopotowska1, Magdalena Maciejewska1, Andrzej Mazur2, Joanna Wietrzyk1,*
1Department of Experimental Oncology, Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw, Poland
2Université Clermont Auvergne, INRA, UNH, Unité de Nutrition Humaine, F-63000 Clermont-Ferrand, France
Download: PDF(878 KB)   HTML
Export: BibTeX | EndNote | Reference Manager | ProCite | RefWorks    
Abstract  

Vitamin D and its analogs are known for their role in the development of breast cancer and in immunomodulation. Our previous studies have shown the pro-metastatic effect of calcitriol and tacalcitol (PRI-2191) in young mice bearing 4T1 breast cancer and the anti-metastatic effect in aged ovariectomized (OVX) mice. Therefore, the aim of our work was to characterize Th17 cell population in young and aged OVX mice bearing 4T1 tumors treated with calcitriol and PRI-2191. The expression of genes typical for Th17 cells was examined in splenocytes, as well as splenocytes differentiated with IL-6 and TGF-β to Th17 cells (iTh17). Expression of genes encoding vitamin D receptor (Vdr) and osteopontin (Spp1) as well as the secretion of IL-17A were evaluated in iTh17 cells. PRI-2191 treatment increased the expression of Rora and Rorc transcription factors, Il17a, Il17re and Il21 in iTh17 cells from young mice. In aged OVX mice this effect was not observed. Increased expression was observed in the case of Vdr and Spp1 genes in iTh17 cells from young mice treated with PRI-2191. What is more, in young mice treated with PRI-2191 the secretion of IL-17A to the culture media by iTh17 cells was increased, whereas in aged OVX mice a significant decrease was noted. Increased expression of Spp1 in young mice treated with PRI-2191 may enhance the differentiation of Th17 cells.

Keywords calcitriol      PRI-2191      Th17      IL-17A      breast cancer      metastasis     
Corresponding Authors: Wietrzyk Joanna   
About author:

Present address: Captor Therapeutics Inc, Wrocław, Poland.

Just Accepted Date: 21 July 2019   Issue Date: 13 March 2020
Service
E-mail this article
E-mail Alert
RSS
Articles by authors
Pawlik Agata
Anisiewicz Artur
Filip-Psurska Beata
Klopotowska Dagmara
Maciejewska Magdalena
Mazur Andrzej
Wietrzyk Joanna
Cite this article:   
Pawlik Agata,Anisiewicz Artur,Filip-Psurska Beata, et al. Divergent Effect of Tacalcitol (PRI-2191) on Th17 Cells in 4T1 Tumor Bearing Young and Old Ovariectomized Mice[J]. Aging and disease, 2020, 11(2): 241-253.
URL:  
http://www.aginganddisease.org/EN/10.14336/AD.2019.0618     OR
Figure 1.  The expression of selected Th17 genes in splenocytes of young (A) and aged (B) mice and in lymph nodes of young mice (C) bearing 4T1 mammary gland tumors. 50 ng of cDNA from untreated splenocytes and 30 ng from lymph nodes were used for a single real-time PCR reaction. Each sample was performed in a triplicate in a single experiment. The real-time PCR reaction was carried out in the Viia 7 device equipped with Viia 7 Software v1.1 software. PCR amplification cycles were performed at 2 min 50ºC, 10 min 95ºC, and then 50 cycles: 95ºC 10 sec, 58ºC 45 sec using specific primers with TaqMan chemistry. Fold-change (RQ) of target cDNA was determined using the ΔΔCt method in reference to the beta-2 microglobulin control gene (B2m; Mm00437762_m1) in the DataAssistTM v3.0.1 software. Number of mice: 5-6/group. Data presented as mean with SD. Statistical analysis: Dunn’s multiple comparison test, *p<0.05.
Figure 2.  Characteristics of CD4+ splenocytes (NT) stimulated with IL-6 and TGF-β (iTh17). The expression of genes typical for Th17 cells in cells from young (A) and aged (B) mice (day 21). Expression of Vdr and Spp1 in iTh17 cells induced from CD4+ splenocytes from young (C) and aged OVX (D) mice. The representative diagrams illustrating the purity of isolated CD4+ splenocytes from young (E) and aged OVX (F) control mice and mice treated with calcitriol. Panel A represents non-stained cells, panel B shows cells stained with anti-CD4 antibody. For a single real-time PCR reaction diluted 1:5 cDNA from previously preamplified stimulated iTh17 splenocytes was used. Each sample was performed in triplicate in a single experiment. Fold-change (RQ) of target cDNA was determined using the ΔΔCt method in reference to the beta-2 microglobulin control gene (B2m; Mm00437762_m1) in the DataAssistTM v3.0.1 software and then calculated as a relative to the values obtained for non-stimulated splenocytes from control mice. Number of mice: 4/group with the exception of control iTh17 cells from aged mice N=3. Data presented as mean with SD. Statistical analysis: Dunn’s multiple comparison test, *p<0.05.
Figure 3.  IL-17A production by iTh17 cells differentiated from CD4+ T lymphocytes (NT) from spleens of young and aged mice bearing 4T1 mammary gland cancer. Number of mice: 5-6/group. Data presented as mean with SD and min-max values. Statistical analysis: Dunn’s multiple comparison test, *p<0.05
Figure 4.  Plasma level of 17β-estradiol in aged OVX mice bearing 4T1 tumors and treated with calcitriol or PRI-2191. Number of mice: 4-5/group. Data presented as mean with SD. Statistical analysis: Dunn’s multiple comparison test, *p<0.05.
Figure 5.  Summary of the effects on iTh17 cells of PRI-2191 (tacalcitol) treatment of young and aged OVX mice bearing 4T1 mammary gland cancer. (A) PRI-2191, by increasing osteopontin level, may indirectly potentiate the differentiation of Th17 cells and the secretion of IL-17 by them. The direct effect of PRI-2191 after binding with VDR on the expression of osteopontin and genes responsible for Th17 differentiation is also considered. (B) In old OVX mice, the observed reduction in the secretion of osteopontin may contribute to reduced secretion of IL-17. Continuous red lines show the effects of PRI-2191 observed in the orthotopic 4T1 mouse breast cancer model, where PRI-2191 stimulated metastasis in young mice and inhibited in old OVX mice. Dotted lines indicate the effects of osteopontin on Th17 cells differentiation (through CD44 or other receptors) observed by other authors in other models (non-cancerous).
[1] Coleman RE, Gregory W, Marshall H, Wilson C, Holen I (2013). The metastatic microenvironment of breast cancer: Clinical implications. Breast, 22: S50-6
[2] DeNardo DG, Johansson M, Coussens LM (2008). Immune cells as mediators of solid tumor metastasis. Cancer Metastasis Rev, 27: 11-8
[3] DeNardo DG, Barreto JB, Andreu P, Vasquez L, Tawfik D, Kolhatkar N, et al. (2009). CD4+ T Cells Regulate Pulmonary Metastasis of Mammary Carcinomas by Enhancing Protumor Properties of Macrophages. Cancer Cell, 16: 91-102
[4] Mantovani A, Marchesi F, Porta C, Sica A, Allavena P (2007). Inflammation and cancer: Breast cancer as a prototype. The Breast, 16: 27-33
[5] Mantovani A, Marchesi F, Porta C, Allavena P, Sica A (2008). Linking inflammation reactions to cancer: Novel targets for therapeutic strategies. Adv Exp Med Biol, 610: 112-27
[6] Su X, Ye J, Hsueh EC, Zhang Y, Hoft DF, Peng G (2010). Tumor Microenvironments Direct the Recruitment and Expansion of Human Th17 Cells J Immunol, 184: 1630-41
[7] Wang S, Li Z, Hu G (2017). Prognostic role of intratumoral IL-17A expression by immunohistochemistry in solid tumors: a meta-analysis. Oncotarget, 8: 66382-91
[8] Chen WC, Lai YH, Chen HY, Guo HR, Su IJ, Chen HHW (2013). Interleukin-17-producing cell infiltration in the breast cancer tumour microenvironment is a poor prognostic factor. Histopathology, 63: 225-33
[9] Cochaud S, Giustiniani J, Thomas C, Laprevotte E, Garbar C, Savoye A-M, et al. (2013). IL-17A is produced by breast cancer TILs and promotes chemoresistance and proliferation through ERK1/2. Sci. Rep., 3: 3456
[10] Asadzadeh Z, Mohammadi H, Safarzadeh E, Hemmatzadeh M, Mahdian-shakib A, Jadidi-Niaragh F, et al. (2017). The paradox of Th17 cell functions in tumor immunity. Cell Immunol, 322: 15-25
[11] Gagliani N, Amezcua Vesely MC, Iseppon A, Brockmann L, Xu H, Palm NW, et al. (2015). TH17 cells transdifferentiate into regulatory T cells during resolution of inflammation. Nature, 523: 221-5
[12] Du JW, Xu KY, Fang LY, Qi XL (2012). Interleukin-17, produced by lymphocytes, promotes tumor growth and angiogenesis in a mouse model of breast cancer. Mol Med Rep, 6: 1099-102
[13] DuPre’ SA, Hunter KW (2007). Murine mammary carcinoma 4T1 induces a leukemoid reaction with splenomegaly: Association with tumor-derived growth factors. Exp Mol Pathol, 82: 12-24
[14] Anisiewicz A, Pawlik A, Filip-Psurska B, Turlej E, Dzimira S, Milczarek M, et al. (2018). Unfavorable effect of calcitriol and its low-calcemic analogs on metastasis of 4T1 mouse mammary gland cancer. Int J Oncol, 52: 103-26
[15] Anisiewicz A, Filip-Psurska B, Pawlik A, Nasulewicz-Goldeman A, Piasecki T, Kowalski K, et al. (2018). Calcitriol Analogues Decrease Lung Metastasis but Impair Bone Metabolism in Aged Ovariectomized Mice Bearing 4T1 Mammary Gland Tumours. Aging Dis, doi:10.14336/AD.2018.0921
[16] Adorini L, Daniel K, Penna G (2006). Vitamin D Receptor Agonists, Cancer and the Immune System: An Intricate Relationship. Curr. Top Med Chem, 6: 1297-301
[17] Liu W, Zhang L, Xu H-J, Li Y, Hu C-M, Yang J-Y, et al. (2018). The Anti-Inflammatory Effects of Vitamin D in Tumorigenesis. Int. J Mol Sci, 19: 2736
[18] Al-Azhri J, Zhang Y, Bshara W, Zirpoli G, McCann SE, Khoury T, et al. (2017). Tumor expression of vitamin D receptor and breast cancer histopathological characteristics and prognosis. Clin Cancer Res, 23: 97-103
[19] Shirazi L, Almquist M, Borgquist S, Malm J, Manjer J (2016). Serum vitamin D (25OHD3) levels and the risk of different subtypes of breast cancer: A nested case-control study. Breast, 28: 184-90
[20] Thanasitthichai S, Chaiwerawattana A, Prasitthipayong A (2015). Association of vitamin D level with clinicopathological features in breast cancer. Asian Pacific J Cancer Prev, 16: 4881-3
[21] Zhang X, Hofmann S, Rack B, Harbeck N, Jeschke U, Sixou S, et al. (2017). Fluorescence Analysis of Vitamin D Receptor Status of Circulating Tumor Cells (CTCS) in Breast Cancer: From Cell Models to Metastatic Patients. Int J Mol Sci, 18: 1318
[22] Ooi LL, Zhou H, Kalak R, Zheng Y, Conigrave AD, Seibel MJ, et al. (2010). Vitamin D deficiency promotes human breast cancer growth in a murine model of bone metastasis. Cancer Res, 70: 1835-44
[23] Williams JD, Aggarwal A, Swami S, Krishnan A V, Ji L, Albertelli MA, et al. (2016). Tumor autonomous effects of Vitamin D deficiency promote breast cancer metastasis. Endocrinology, 157: 1341-7
[24] Lu G, Li J, Chu J, Jin S, Fu Z, Miao D, et al. (2017). 1,25(OH)2D3 deficiency increases TM40D tumor growth in bone and accelerates tumor-induced bone destruction in a breast cancer bone metastasis model. Biomed. Pharmacother, 95: 1033-9
[25] Chen L, Yang R, Qiao W, Yuan X, Wang S, Goltzman D, et al. (2018). 1,25-Dihydroxy vitamin D prevents tumorigenesis by inhibiting oxidative stress and inducing tumor cellular senescence in mice. Int J Cancer, 143: 368-82
[26] Milczarek M, Chodyński M, Filip-Psurska B, Martowicz A, Krupa M, Krajewski K, et al. (2013). Synthesis and Biological Activity of Diastereomeric and Geometric Analogs of Calcipotriol, PRI-2202 and PRI-2205, Against Human HL-60 Leukemia and MCF-7 Breast Cancer Cells. Cancers, 5: 1355-78
[27] Chiang K-C, Yeh C-N, Yeh T-S, Juang H-H, Chen L-W, Kuo S-F, et al. (2018). MART-10, a 1alpha, 25(OH)2D3 Analog, Potently Represses Metastasis of ER(+) Breast Cancer Cells with VEGF-A Overexpression. Anticancer Res, 38: 3879-87
[28] Wilmanski T, Barnard A, Parikh MR, Kirshner J, Buhman K, Burgess J, et al. (2016). 1α,25-Dihydroxyvitamin D Inhibits the Metastatic Capability of MCF10CA1a and MDA-MB-231 Cells in an In Vitro Model of Breast to Bone Metastasis. Nutr Cancer, 68: 1202-9
[29] Estébanez N, Gómez-Acebo I, Palazuelos C, Llorca J, Dierssen-Sotos T (2018). Vitamin D exposure and Risk of Breast Cancer: a meta-analysis. Sci Rep, 8: 9039
[30] Manson JE, Cook NR, Lee I-M, Christen W, Bassuk SS, Mora S, et al. (2018). Vitamin D Supplements and Prevention of Cancer and Cardiovascular Disease. N. Engl J Med, 380: NEJMoa1809944
[31] Cao Y, Du Y, Liu F, Feng Y, Cheng S, Guan S, et al. (2018). Vitamin D aggravates breast cancer by inducing immunosuppression in the tumor bearing mouse. Immunotherapy, 10: 555-66
[32] Ajibade AA, Kirk JS, Karasik E, Gillard B, Moser MT, Johnson CS, et al. (2014). Early Growth Inhibition Is Followed by Increased Metastatic Disease with Vitamin D (Calcitriol) Treatment in the TRAMP Model of Prostate Cancer. PLoS One, 9: e89555
[33] Pawlik A, Anisiewicz A, Filip-Psurska B, Nowak M, Turlej E, Trynda J, et al. (2018). Calcitriol and Its Analogs Establish the Immunosuppressive Microenvironment That Drives Metastasis in 4T1 Mouse Mammary Gland Cancer. Int J Mol Sci, 19: 2116
[34] Kudlacek S, Willvonseder R, Stohlawetz P, Hahn P, Pietschmann P (2000). Immunology and aging. Aging Male, 3: 137-42
[35] Wietrzyk J, Pełczyńska M, Madej J, Dzimira S, Kuśnierczyk H, Kutner A, et al. (2004). Toxicity and antineoplastic effect of (24R)-1,24-dihydroxyvitamin D 3 (PRI-2191). Steroids, 69: 629-35
[36] Wietrzyk J, Chodyński M, Fitak H, Wojdat E, Kutner A, Opolski A (2007). Antitumor properties of diastereomeric and geometric analogs of vitamin D3. Anticancer. Drugs, 18: 447-57
[37] Tao K, Fang M, Alroy J, Sahagian GG (2008). Imagable 4T1 model for the study of late stage breast cancer. BMC Cancer, 8: 228
[38] Qian X, Gu L, Ning H, Zhang Y, Hsueh EC, Fu M, et al. (2013). Increased Th17 Cells in the Tumor Microenvironment Is Mediated by IL-23 via Tumor-Secreted Prostaglandin E2. J Immunol, 190: 5894-902
[39] Nam J-S, Terabe M, Kang M-J, Chae H, Voong N, Yang Y -a, et al. (2008). Transforming Growth Factor Subverts the Immune System into Directly Promoting Tumor Growth through Interleukin-17. Cancer Res, 68: 3915-23
[40] Nurieva R, Yang XO, Martinez G, Zhang Y, Panopoulos AD, Ma L, et al. (2007). Essential autocrine regulation by IL-21 in the generation of inflammatory T cells. Nature, 448: 480-3
[41] Yang L, Anderson DE, Baecher-Allan C, Hastings WD, Bettelli E, Oukka M, et al. (2008). IL-21 and TGF-β are required for differentiation of human TH17 cells. Nature, 454: 350-2
[42] Chung Y, Chang SH, Martinez GJ, Yang XO, Nurieva R, Kang HS, et al. (2009). Critical Regulation of Early Th17 Cell Differentiation by Interleukin-1 Signaling. Immunity, 30: 576-87
[43] Yang XO, Pappu BP, Nurieva R, Akimzhanov A, Kang HS, Chung Y, et al. (2008). T Helper 17 Lineage Differentiation Is Programmed by Orphan Nuclear Receptors RORα and RORγ. Immunity, 28: 29-39
[44] Castro G, Liu X, Ngo K, De Leon-Tabaldo A, Zhao S, Luna-Roman R, et al. (2017). RORγt and RORα signature genes in human Th17 cells. PLoS One, 12: e0181868
[45] Song Y, Yang JM (2017). Role of interleukin (IL)-17 and T-helper (Th)17 cells in cancer. Biochem. Biophys Res Commun, 493: 1-8
[46] Gulubova M, Ananiev J, Ignatova M, Halacheva K (2016). Pro-Tumor and Anti-Tumor Functions of IL-17 and of TH17 Cells in Tumor Microenvironment. Acta Medica Bulg, 43: 68-79
[47] Hayes CE, Hubler SL, Moore JR, Barta LE, Praska CE, Nashold FE (2015). Vitamin D Actions on CD4+ T Cells in Autoimmune Disease. Front Immunol, 6: 1-22
[48] Tang J, Zhou R, Luger D, Zhu W, Silver PB, Grajewski RS, et al. (2009). Calcitriol suppresses antiretinal autoimmunity through inhibitory effects on the Th17 effector response. J Immunol, 182: 4624-32
[49] Smolders J, Thewissen M, Peelen E, Menheere P, Tervaert JWC, Damoiseaux J, et al. (2009). Vitamin D status is positively correlated with regulatory T cell function in patients with multiple sclerosis. PLoS One, 4: e6635
[50] Smolders J, Peelen E, Thewissen M, Cohen Tervaert JW, Menheere P, Hupperts R, et al. (2010). Safety and T cell modulating effects of high dose vitamin D3 supplementation in multiple sclerosis. PLoS One, 5: e15235
[51] Nashold FE, Spach KM, Spanier JA, Hayes CE (2009). Estrogen Controls Vitamin D3-Mediated Resistance to Experimental Autoimmune Encephalomyelitis by Controlling Vitamin D3 Metabolism and Receptor Expression. J. Immunol, 183: 3672-81
[52] Chen R-Y, Fan Y-M, Zhang Q, Liu S, Li Q, Ke G-L, et al. (2015). Estradiol Inhibits Th17 Cell Differentiation through Inhibition of RORγT Transcription by Recruiting the ERα/REA Complex to Estrogen Response Elements of the RORγT Promoter. J Immunol, 194: 4019-28
[53] Pang H, Lu H, Song H, Meng Q, Zhao Y, Liu N, et al. (2013). Prognostic values of osteopontin-c, E-cadherin and β-catenin in breast cancer. Cancer Epidemiol, 37: 985-92
[54] Shan M, Yuan X, Song LZ, Roberts L, Zarinkamar N, Seryshev A, et al. (2012). Cigarette smoke induction of osteopontin (SPP1) mediates TH17 inflammation in human and experimental emphysema. Sci Transl Med, 4: 117ra9
[55] Murugaiyan G, Mittal A, Weiner HL (2008). Increased osteopontin expression in dendritic cells amplifies IL-17 production by CD4+ T cells in experimental autoimmune encephalomyelitis and in multiple sclerosis. J Immunol, 181: 7480-8
[56] Zheng Y, Wang Z, Deng L, Yuan X, Ma Y, Zhang G, et al. (2012). Osteopontin promotes inflammation in patients with acute coronary syndrome through its activity on IL-17 producing cells. Eur. J Immunol, 42: 2803-14
[57] Cao Z, Cheng W, Niu X, Zhao Q, Wu T, Li C, et al. (2017). IFN-β regulates Th17 differentiation partly through the inhibition of osteopontin in experimental autoimmune encephalomyelitis. Mol Immunol, 93: 20-30
[58] Chang PL, Prince CW (1991). 1α,25-Dihydroxyvitamin D3 Stimulates Synthesis and Secretion of Nonphosphorylated Osteopontin (Secreted Phosphoprotein 1) in Mouse JB6 Epidermal Cells. Cancer Res, 51: 2144-50
[59] Lau WL, Leaf EM, Hu MC, Takeno MM, Kuro-o M, Moe OW, et al. (2012). Vitamin D receptor agonists increase klotho and osteopontin while decreasing aortic calcification in mice with chronic kidney disease fed a high phosphate diet. Kidney Int, 82: 1261-70
[60] Huber M, Heink S, Pagenstecher A, Reinhard K, Ritter J, Visekruna A, et al. (2013). IL-17A secretion by CD8+ T cells supports Th17-mediated autoimmune encephalomyelitis. J Clin Invest, 123: 247-60
[61] Duan M-C, Huang Y, Zhong X-N, Tang H-J (2012). Th17 Cell Enhances CD8 T-Cell Cytotoxicity via IL-21 Production in Emphysema Mice. Mediators Inflamm, 2012: 1-12
[62] Li J, Yue L, Wang H, Liu C, Liu H, Tao J, et al. (2016). Th17 Cells Exhibit Antitumor Effects in MDS Possibly through Augmenting Functions of CD8+ T Cells. J Immunol Res, 2016: 1-14
[1] Supplementary data Download
[1] Yu Gu, Junhua Zhang, Zhirui Zhou, Di Liu, Hongcheng Zhu, Junmiao Wen, Xinyan Xu, Tianxiang Chen, Min Fan. Metastasis Patterns and Prognosis of Octogenarians with NSCLC: A Population-based Study[J]. Aging and disease, 2020, 11(1): 82-92.
[2] Artur Anisiewicz, Beata Filip-Psurska, Agata Pawlik, Anna Nasulewicz-Goldeman, Tomasz Piasecki, Konrad Kowalski, Magdalena Maciejewska, Joanna Jarosz, Joanna Banach, Diana Papiernik, Andrzej Mazur, Andrzej Kutner, Jeanette A Maier, Joanna Wietrzyk. Calcitriol Analogues Decrease Lung Metastasis but Impair Bone Metabolism in Aged Ovariectomized Mice Bearing 4T1 Mammary Gland Tumours[J]. Aging and disease, 2019, 10(5): 977-991.
[3] Xu Limin, Li Liqin, Li Jing, Li Hongwei, Shen Qibin, Ping Jinliang, Ma Zhihong, Zhong Jing, Dai Licheng. Overexpression of miR-1260b in Non-small Cell Lung Cancer is Associated with Lymph Node Metastasis[J]. Aging and disease, 2015, 6(6): 478-485.
Viewed
Full text


Abstract

Cited

  Shared   
Copyright © 2014 Aging and Disease, All Rights Reserved.
Address: Aging and Disease Editorial Office 3400 Camp Bowie Boulevard Fort Worth, TX76106 USA
Fax: (817) 735-0408 E-mail: editorial@aginganddisease.org
Powered by Beijing Magtech Co. Ltd