Selective anti-CXCR2 receptor blockade by AZD5069 inhibits CXCL8-mediated pro-tumorigenic activity in human thyroid cancer cells in vitro.
AZD5069
CXCL8
CXCR2 receptor
Thyroid cancer
Journal
Journal of endocrinological investigation
ISSN: 1720-8386
Titre abrégé: J Endocrinol Invest
Pays: Italy
ID NLM: 7806594
Informations de publication
Date de publication:
20 Jun 2024
20 Jun 2024
Historique:
received:
30
04
2024
accepted:
06
06
2024
medline:
20
6
2024
pubmed:
20
6
2024
entrez:
20
6
2024
Statut:
aheadofprint
Résumé
Thyroid cancer is the most common endocrine malignancy. Current therapies are successful, however some patients progress to therapeutically refractive disease. The immunotherapeutic potential of the CXCL8-chemokine/CXCR2-chemokine-receptor system is currently being explored in numerous human cancers. This study aimed to evaluate if the targeting of CXCR2 by its selective antagonist, AZD5069, could modulate CXCL8-mediated pro-tumorigenic effects in thyroid-cancer (TC) cells in vitro. Normal human primary thyroid cells (NHT) and TC cell lines TPC-1 (RET/PTC), BCPAP, 8505C and 8305C (BRAFV600e) were treated with AZD5069 (100 pM-10 µM) over a time-course. Viability and proliferation were assessed by WST-1 and crystal violet assays. CXCL8 and CXCR2 mRNA were evaluated by RT-PCR. CXCL8-protein concentrations were measured in cell culture supernatants by ELISA. CXCR2 on cell surface was evaluated by flow-cytometry. Cell-migration was assessed by trans-well-migration chamber-system. AZD5069 exerted negligible effects on cell proliferation or viability. AZD5069 significantly reduced CXCR2, (but not CXCL8) mRNAs in all cell types. CXCR2 was reduced on the membrane of some TC cell lines. A significant reduction of the CXCL8 secretion was found in TPC-1 cells (basal-secretion) and NHT (TNFα-induced secretion). AZD5069 significantly reduced basal and CXCL8-induced migration in NHT and different TC cells. Our findings confirm the involvement of the CXCL8/CXCR2-axis in promoting pro-tumorigenic effects in TC cells, further demonstrating its immunotherapeutic significance in human cancer.
Sections du résumé
BACKGROUND
BACKGROUND
Thyroid cancer is the most common endocrine malignancy. Current therapies are successful, however some patients progress to therapeutically refractive disease. The immunotherapeutic potential of the CXCL8-chemokine/CXCR2-chemokine-receptor system is currently being explored in numerous human cancers. This study aimed to evaluate if the targeting of CXCR2 by its selective antagonist, AZD5069, could modulate CXCL8-mediated pro-tumorigenic effects in thyroid-cancer (TC) cells in vitro.
METHODS
METHODS
Normal human primary thyroid cells (NHT) and TC cell lines TPC-1 (RET/PTC), BCPAP, 8505C and 8305C (BRAFV600e) were treated with AZD5069 (100 pM-10 µM) over a time-course. Viability and proliferation were assessed by WST-1 and crystal violet assays. CXCL8 and CXCR2 mRNA were evaluated by RT-PCR. CXCL8-protein concentrations were measured in cell culture supernatants by ELISA. CXCR2 on cell surface was evaluated by flow-cytometry. Cell-migration was assessed by trans-well-migration chamber-system.
RESULTS
RESULTS
AZD5069 exerted negligible effects on cell proliferation or viability. AZD5069 significantly reduced CXCR2, (but not CXCL8) mRNAs in all cell types. CXCR2 was reduced on the membrane of some TC cell lines. A significant reduction of the CXCL8 secretion was found in TPC-1 cells (basal-secretion) and NHT (TNFα-induced secretion). AZD5069 significantly reduced basal and CXCL8-induced migration in NHT and different TC cells.
CONCLUSIONS
CONCLUSIONS
Our findings confirm the involvement of the CXCL8/CXCR2-axis in promoting pro-tumorigenic effects in TC cells, further demonstrating its immunotherapeutic significance in human cancer.
Identifiants
pubmed: 38900374
doi: 10.1007/s40618-024-02410-6
pii: 10.1007/s40618-024-02410-6
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : Ministero della Salute
ID : Ricerca Corrente
Informations de copyright
© 2024. The Author(s).
Références
Davies L, Welch HG (2006) Increasing incidence of thyroid cancer in the United States, 1973–2002. JAMA 295(18):2164–2167
pubmed: 16684987
doi: 10.1001/jama.295.18.2164
McLeod DSA, Zhang L, Durante C, Cooper DS (2019) Contemporary debates in adult papillary thyroid cancer management. Endocr Rev 40(6):1481–1499
pubmed: 31322698
doi: 10.1210/er.2019-00085
Network CGAR (2014) Integrated genomic characterization of papillary thyroid carcinoma. Cell 159(3):676–690
doi: 10.1016/j.cell.2014.09.050
Pani F, Caria P, Yasuda Y, Makoto M, Mariotti S, Leenhardt L et al (2022) The immune landscape of papillary thyroid cancer in the context of autoimmune thyroiditis. Cancers 14(17):4287
pubmed: 36077831
pmcid: 9454449
doi: 10.3390/cancers14174287
Pizzato M, Li M, Vignat J, Laversanne M, Singh D, La Vecchia C et al (2022) The epidemiological landscape of thyroid cancer worldwide: GLOBOCAN estimates for incidence and mortality rates in 2020. Lancet Diabetes Endocrinol 10(4):264–272
pubmed: 35271818
doi: 10.1016/S2213-8587(22)00035-3
Haugen BR, Sawka AM, Alexander EK, Bible KC, Caturegli P, Doherty GM et al (2017) American thyroid association guidelines on the management of thyroid nodules and differentiated thyroid cancer task force review and recommendation on the proposed renaming of encapsulated follicular variant papillary thyroid carcinoma without invasion to noninvasive follicular thyroid neoplasm with papillary-like nuclear features. Thyroid 27(4):481–483
pubmed: 28114862
doi: 10.1089/thy.2016.0628
Rahib L, Smith BD, Aizenberg R, Rosenzweig AB, Fleshman JM, Matrisian LM (2014) Projecting cancer incidence and deaths to 2030: the unexpected burden of thyroid, liver, and pancreas cancers in the United States. Cancer Res 74(11):2913–2921
pubmed: 24840647
doi: 10.1158/0008-5472.CAN-14-0155
Cunha LL, Ward LS (2022) Translating the immune microenvironment of thyroid cancer into clinical practice. Endocr Relat Cancer 29(6):R67–R83
pubmed: 35289765
doi: 10.1530/ERC-21-0414
Menicali E, Guzzetti M, Morelli S, Moretti S, Puxeddu E (2020) Immune landscape of thyroid cancers: new insights. Front Endocrinol (Lausanne) 11:637826
pubmed: 33986723
doi: 10.3389/fendo.2020.637826
Febrero B, Ruiz-Manzanera JJ, Ros-Madrid I, Hernández AM, Orenes-Piñero E, Rodríguez JM (2024) Tumor microenvironment in thyroid cancer: Immune cells, patterns, and novel treatments. Head Neck 46(6):1486–1499
pubmed: 38380767
doi: 10.1002/hed.27695
Rotondi M, Coperchini F, Latrofa F, Chiovato L (2018) Role of chemokines in thyroid cancer microenvironment: Is CXCL8 the main player? Front Endocrinol (Lausanne) 9:314
pubmed: 29977225
doi: 10.3389/fendo.2018.00314
Xie K (2001) Interleukin-8 and human cancer biology. Cytokine Growth Factor Rev 12(4):375–391
pubmed: 11544106
doi: 10.1016/S1359-6101(01)00016-8
Fang W, Ye L, Shen L, Cai J, Huang F, Wei Q et al (2014) Tumor-associated macrophages promote the metastatic potential of thyroid papillary cancer by releasing CXCL8. Carcinogenesis 35(8):1780–1787
pubmed: 24608042
doi: 10.1093/carcin/bgu060
Sanmamed MF, Carranza-Rua O, Alfaro C, Oñate C, Martín-Algarra S, Perez G et al (2014) Serum interleukin-8 reflects tumor burden and treatment response across malignancies of multiple tissue origins. Clin Cancer Res 20(22):5697–5707
pubmed: 25224278
doi: 10.1158/1078-0432.CCR-13-3203
Jurcevic S, Humfrey C, Uddin M, Warrington S, Larsson B, Keen C (2015) The effect of a selective CXCR2 antagonist (AZD5069) on human blood neutrophil count and innate immune functions. Br J Clin Pharmacol 80(6):1324–1336
pubmed: 26182832
pmcid: 4693488
doi: 10.1111/bcp.12724
Uddin M, Betts C, Robinson I, Malmgren A, Humfrey C (2017) The chemokine CXCR2 antagonist (AZD5069) preserves neutrophil-mediated host immunity in non-human primates. Haematologica 102(2):e65–e68
pubmed: 27742769
pmcid: 5286957
doi: 10.3324/haematol.2016.152371
Sody S, Uddin M, Grüneboom A, Görgens A, Giebel B, Gunzer M et al (2019) Distinct spatio-temporal dynamics of tumor-associated neutrophils in small tumor lesions. Front Immunol 10:1419
pubmed: 31293583
pmcid: 6603174
doi: 10.3389/fimmu.2019.01419
Saintigny P, Massarelli E, Lin S, Ahn YH, Chen Y, Goswami S et al (2013) CXCR2 expression in tumor cells is a poor prognostic factor and promotes invasion and metastasis in lung adenocarcinoma. Cancer Res 73(2):571–582
pubmed: 23204236
doi: 10.1158/0008-5472.CAN-12-0263
Yang L, Huang J, Ren X, Gorska AE, Chytil A, Aakre M et al (2008) Abrogation of TGF beta signaling in mammary carcinomas recruits Gr-1+CD11b+ myeloid cells that promote metastasis. Cancer Cell 13(1):23–35
pubmed: 18167337
pmcid: 2245859
doi: 10.1016/j.ccr.2007.12.004
Steele CW, Karim SA, Leach JDG, Bailey P, Upstill-Goddard R, Rishi L et al (2016) CXCR2 inhibition profoundly suppresses metastases and augments immunotherapy in pancreatic ductal adenocarcinoma. Cancer Cell 29(6):832–845
pubmed: 27265504
pmcid: 4912354
doi: 10.1016/j.ccell.2016.04.014
Guo F, Long L, Wang J, Wang Y, Liu Y, Wang L et al (2019) Insights on CXC chemokine receptor 2 in breast cancer: an emerging target for oncotherapy. Oncol Lett 18(6):5699–5708
pubmed: 31788042
pmcid: 6865047
Liotti F, De Pizzol M, Allegretti M, Prevete N, Melillo RM (2017) Multiple anti-tumor effects of Reparixin on thyroid cancer. Oncotarget 8(22):35946–35961
pubmed: 28415590
pmcid: 5482629
doi: 10.18632/oncotarget.16412
Safarulla S, Madan A, Xing F, Chandrasekaran A (2022) CXCR2 mediates distinct neutrophil behavior in brain metastatic breast tumor. Cancers 14(3):515
pubmed: 35158784
pmcid: 8833752
doi: 10.3390/cancers14030515
Coperchini F, Pignatti P, Carbone A, Bongianino R, Di Buduo CA, Leporati P et al (2016) TNF-α increases the membrane expression of the chemokine receptor CCR6 in thyroid tumor cells, but not in normal thyrocytes: potential role in the metastatic spread of thyroid cancer. Tumour Biol 37(4):5569–5575
pubmed: 26577851
doi: 10.1007/s13277-015-4418-7
Watz H, Uddin M, Pedersen F, Kirsten A, Goldmann T, Stellmacher F et al (2017) Effects of the CXCR2 antagonist AZD5069 on lung neutrophil recruitment in asthma. Pulm Pharmacol Ther 45:121–123
pubmed: 28549850
doi: 10.1016/j.pupt.2017.05.012
Cullberg M, Arfvidsson C, Larsson B, Malmgren A, Mitchell P, Wählby Hamrén U et al (2018) Pharmacokinetics of the oral selective CXCR2 antagonist azd5069: a summary of eight phase I studies in healthy volunteers. Drugs R D 18(2):149–159
pubmed: 29856004
pmcid: 5995788
doi: 10.1007/s40268-018-0236-x
Crowley LC, Christensen ME, Waterhouse NJ (2016) Measuring survival of adherent cells with the colony-forming assay. Cold Spring Harb Protoc. https://doi.org/10.1101/pdb.prot087171
doi: 10.1101/pdb.prot087171
pubmed: 27934691
Coperchini F, Croce L, Denegri M, Pignatti P, Agozzino M, Netti GS et al (2020) Adverse effects of in vitro GenX exposure on rat thyroid cell viability, DNA integrity and thyroid-related genes expression. Environ Pollut 264:114778
pubmed: 32417585
doi: 10.1016/j.envpol.2020.114778
Abbonante V, Gruppi C, Rubel D, Gross O, Moratti R, Balduini A (2013) Discoidin domain receptor 1 protein is a novel modulator of megakaryocyte-collagen interactions. J Biol Chem 288(23):16738–16746
pubmed: 23530036
pmcid: 3675607
doi: 10.1074/jbc.M112.431528
Rotondi M, Coperchini F, Pignatti P, Sideri R, Groppelli G, Leporati P et al (2013) Interferon-γ and tumor necrosis factor-α sustain secretion of specific CXC chemokines in human thyrocytes: a first step toward a differentiation between autoimmune and tumor-related inflammation? J Clin Endocrinol Metab 98(1):308–313
pubmed: 23118425
doi: 10.1210/jc.2012-2555
Coperchini F, Croce L, Denegri M, Awwad O, Ngnitejeu ST, Muzza M et al (2019) The BRAF-inhibitor PLX4720 inhibits CXCL8 secretion in BRAFV600E mutated and normal thyroid cells: a further anti-cancer effect of BRAF-inhibitors. Sci Rep 9(1):4390
pubmed: 30867499
pmcid: 6416278
doi: 10.1038/s41598-019-40818-w
Coperchini F, Croce L, Denegri M, Awwad O, Ngnitejeu ST, Magri F et al (2019) The anti-cancer effects of phenformin in thyroid cancer cell lines and in normal thyrocytes. Oncotarget 10(60):6432–6443
pubmed: 31741708
pmcid: 6849649
doi: 10.18632/oncotarget.27266
Awwad O, Coperchini F, Pignatti P, Denegri M, Massara S, Croce L et al (2018) The AMPK-activator AICAR in thyroid cancer: effects on CXCL8 secretion and on CXCL8-induced neoplastic cell migration. J Endocrinol Invest 41:1275–1282
pubmed: 29546654
doi: 10.1007/s40618-018-0862-8
David JM, Dominguez C, Hamilton DH, Palena C (2016) The IL-8/IL-8R axis: a double agent in tumor immune resistance. Vaccines 4(3):22
pubmed: 27348007
pmcid: 5041016
doi: 10.3390/vaccines4030022
Alfaro C, Sanmamed MF, Rodríguez-Ruiz ME, Teijeira Á, Oñate C, González Á et al (2017) Interleukin-8 in cancer pathogenesis, treatment and follow-up. Cancer Treat Rev 60:24–31
pubmed: 28866366
doi: 10.1016/j.ctrv.2017.08.004
Fousek K, Horn LA, Palena C (2021) Interleukin-8: a chemokine at the intersection of cancer plasticity, angiogenesis, and immune suppression. Pharmacol Ther 219:107692
pubmed: 32980444
doi: 10.1016/j.pharmthera.2020.107692
Coperchini F, Croce L, Marinò M, Chiovato L, Rotondi M (2019) Role of chemokine receptors in thyroid cancer and immunotherapy. Endocr Relat Cancer 26(8):R465–R478
pubmed: 31146261
doi: 10.1530/ERC-19-0163
Maier T, Güell M, Serrano L (2009) Correlation of mRNA and protein in complex biological samples. FEBS Lett 583(24):3966–3973
pubmed: 19850042
doi: 10.1016/j.febslet.2009.10.036
O’Byrne PM, Metev H, Puu M, Richter K, Keen C, Uddin M et al (2016) Efficacy and safety of a CXCR2 antagonist, AZD5069, in patients with uncontrolled persistent asthma: a randomised, double-blind, placebo-controlled trial. Lancet Respir Med 4(10):797–806
pubmed: 27574788
doi: 10.1016/S2213-2600(16)30227-2
Mårdh CK, Root J, Uddin M, Stenvall K, Malmgren A, Karabelas K et al (2017) Targets of neutrophil influx and weaponry: therapeutic opportunities for chronic obstructive airway disease. J Immunol Res 2017:5273201
pubmed: 28596972
pmcid: 5449733
doi: 10.1155/2017/5273201
Cristinziano L, Modestino L, Loffredo S, Varricchi G, Braile M, Ferrara AL et al (2020) Anaplastic thyroid cancer cells induce the release of mitochondrial extracellular DNA traps by viable neutrophils. J Immunol 204(5):1362–1372
pubmed: 31959732
doi: 10.4049/jimmunol.1900543
Wislez M, Rabbe N, Marchal J, Milleron B, Crestani B, Mayaud C et al (2003) Hepatocyte growth factor production by neutrophils infiltrating bronchioloalveolar subtype pulmonary adenocarcinoma: role in tumor progression and death. Cancer Res 63(6):1405–1412
pubmed: 12649206
Liu Q, Li A, Tian Y, Wu JD, Liu Y, Li T et al (2016) The CXCL8-CXCR1/2 pathways in cancer. Cytokine Growth Factor Rev 31:61–71
pubmed: 27578214
pmcid: 6142815
doi: 10.1016/j.cytogfr.2016.08.002
Wang J, Jia Y, Wang N, Zhang X, Tan B, Zhang G et al (2014) The clinical significance of tumor-infiltrating neutrophils and neutrophil-to-CD8+ lymphocyte ratio in patients with resectable esophageal squamous cell carcinoma. J Transl Med 12:7
pubmed: 24397835
pmcid: 3895663
doi: 10.1186/1479-5876-12-7
Naito Y, Yamamoto Y, Sakamoto N, Shimomura I, Kogure A, Kumazaki M et al (2019) Cancer extracellular vesicles contribute to stromal heterogeneity by inducing chemokines in cancer-associated fibroblasts. Oncogene 38(28):5566–5579
pubmed: 31147602
pmcid: 6755971
doi: 10.1038/s41388-019-0832-4
Rapoport BL, Steel HC, Theron AJ, Smit T, Anderson R (2020) Role of the neutrophil in the pathogenesis of advanced cancer and impaired responsiveness to therapy. Molecules 25(7):1618
pubmed: 32244751
pmcid: 7180559
doi: 10.3390/molecules25071618
Pedersen F, Waschki B, Marwitz S, Goldmann T, Kirsten A, Malmgren A et al (2018) Neutrophil extracellular trap formation is regulated by CXCR2 in COPD neutrophils. Eur Respir J 51(4):1700970
pubmed: 29449427
doi: 10.1183/13993003.00970-2017
Uddin M, Watz H, Malmgren A, Pedersen F (2019) NETopathic inflammation in chronic obstructive pulmonary disease and severe asthma. Front Immunol 10:47
pubmed: 30804927
pmcid: 6370641
doi: 10.3389/fimmu.2019.00047
Xiong G, Chen Z, Liu Q, Peng F, Zhang C, Cheng M et al (2024) CD276 regulates the immune escape of esophageal squamous cell carcinoma through CXCL1-CXCR2 induced NETs. J Immunother Cancer 12(5):e008662
pubmed: 38724465
pmcid: 11086492
doi: 10.1136/jitc-2023-008662
Baggiolini M (1998) Chemokines and leukocyte traffic. Nature 392(6676):565–568
pubmed: 9560152
doi: 10.1038/33340
Vandercappellen J, Van Damme J, Struyf S (2008) The role of CXC chemokines and their receptors in cancer. Cancer Lett 267(2):226–244
pubmed: 18579287
doi: 10.1016/j.canlet.2008.04.050
Rotondi M, Coperchini F, Pignatti P, Magri F, Chiovato L (2015) Metformin reverts the secretion of CXCL8 induced by TNF-α in primary cultures of human thyroid cells: an additional indirect anti-tumor effect of the drug. J Clin Endocrinol Metab 100(3):E427–E432
pubmed: 25590211
doi: 10.1210/jc.2014-3045
Zhang P, Guan H, Yuan S, Cheng H, Zheng J, Zhang Z et al (2022) Targeting myeloid derived suppressor cells reverts immune suppression and sensitizes BRAF-mutant papillary thyroid cancer to MAPK inhibitors. Nat Commun 13(1):1588
pubmed: 35332119
pmcid: 8948260
doi: 10.1038/s41467-022-29000-5
Leslie J, Mackey JBG, Jamieson T, Ramon-Gil E, Drake TM, Fercoq F et al (2022) CXCR2 inhibition enables NASH-HCC immunotherapy. Gut 71(10):2093–2106
pubmed: 35477863
doi: 10.1136/gutjnl-2021-326259