Cigarette smoke-induced emphysema exhausts early cytotoxic CD8 + T cell responses against nascent lung cancer cells

Journal of Immunology

Volumn 201, Issue 5, 2018, Pages 1558-1569

  Kerdidani, Dimitra ^a,b   Magkouta, Sophia ^b   Chouvardas, Panagiotis ^a,b   Karavana, Vassiliki ^b   Glynos, Konstantinos ^b   Roumelioti, Fani ^a   Zakynthinos, Spyros ^b   Wauters, Els ^c,d   Janssens, Wim ^c,d   Lambrechts, Diether ^e,f   Kollias, George ^a,b   Tsoumakidou, Maria ^a  

^a BIOMEDICAL SCIENCES RESEARCH CENTER ALEXANDER FLEMING   (Greece)

^b UNIVERSITY OF ATHENS MEDICAL SCHOOL   (Greece)

^c UNIVERSITY HOSPITALS LEUVEN   (Belgium)

^d UNIVERSITY OF LEUVEN   (Belgium)

^e DEPARTMENT OF PLANT SYSTEMS BIOLOGY   (Belgium)

^f UNIVERSITY OF LEUVEN   (Belgium)

Author keywords

[No Author keywords available]

Indexed keywords

GLYCOPROTEIN P 15095; MAJOR HISTOCOMPATIBILITY ANTIGEN CLASS 2; PROGRAMMED DEATH 1 LIGAND 1;

ADOPTIVE TRANSFER; ANIMAL CELL; ANIMAL EXPERIMENT; ANIMAL MODEL; ANIMAL TISSUE; ARTICLE; CANCER INHIBITION; CD8+ T LYMPHOCYTE; CELL EXPANSION; CELL FUNCTION; CELL INFILTRATION; CELL SELECTION; CELLULAR IMMUNITY; CIGARETTE SMOKE-INDUCED EMPHYSEMA; CYTOTOXIC T LYMPHOCYTE; DOWN REGULATION; IMMUNOCOMPETENCE; IMMUNOREGULATION; LUNG CANCER; LYMPHOCYTE ACTIVATION; LYMPHOCYTE TRANSFER; MOUSE; MOUSE MODEL; MYELOID DENDRITIC CELL; NONHUMAN; OXIDATIVE STRESS; PRIORITY JOURNAL; PROTEIN EXPRESSION; TUMOR MICROENVIRONMENT; UPREGULATION; ANIMAL; CIGARETTE SMOKING; IMMUNOLOGY; LUNG EMPHYSEMA; LUNG TUMOR; PATHOLOGY; PATHOPHYSIOLOGY; TRANSGENIC MOUSE; TRANSPLANTATION;

ADOPTIVE TRANSFER; ANIMALS; CD8-POSITIVE T-LYMPHOCYTES; CIGARETTE SMOKING; LUNG NEOPLASMS; MICE; MICE, TRANSGENIC; PULMONARY EMPHYSEMA;

EID: 85052446122     PISSN: 00221767     EISSN: 15506606     Source Type: Journal    
DOI: 10.4049/jimmunol.1700700     Document Type: Article

References (80)

1. Global Initiative for Chronic Obstructive Lung Disease. 2017. GOLD 2017 Global Strategy for the Diagnosis, Management and Prevention of COPD. Available from: https://goldcopd.org. Accessed July 19, 2018.
2. Punturieri, A., E. Szabo, T. L. Croxton, S. D. Shapiro, and S. M. Dubinett. 2009. Lung cancer and chronic obstructive pulmonary disease: needs and opportunities for integrated research. J. Natl. Cancer Inst. 101: 554-559.
3. Young, R. P., R. J. Hopkins, T. Christmas, P. N. Black, P. Metcalf, and G. D. Gamble. 2009. COPD prevalence is increased in lung cancer, independent of age, sex and smoking history. Eur. Respir. J. 34: 380-386.
4. Lange, P., J. Nyboe, M. Appleyard, G. Jensen, and P. Schnohr. 1990. Ventilatory function and chronic mucus hypersecretion as predictors of death from lung cancer. Am. Rev. Respir. Dis. 141: 613-617.
5. Shaheen, S. O., J. A. Sterne, J. S. Tucker, and C. D. Florey. 1998. Birth weight, childhood lower respiratory tract infection, and adult lung function. Thorax 53: 549-553.
6. Wilson, D. O., J. L. Weissfeld, A. Balkan, J. G. Schragin, C. R. Fuhrman, S. N. Fisher, J. Wilson, J. K. Leader, J. M. Siegfried, S. D. Shapiro, and F. C. Sciurba. 2008. Association of radiographic emphysema and airflow obstruction with lung cancer. Am. J. Respir. Crit. Care Med. 178: 738-744.
7. Zulueta, J. J., J. P. Wisnivesky, C. I. Henschke, R. Yip, A. O. Farooqi, D. I. McCauley, M. Chen, D. M. Libby, J. P. Smith, M. W. Pasmantier, and D. F. Yankelevitz. 2012. Emphysema scores predict death from COPD and lung cancer. Chest 141: 1216-1223.
8. Oelsner, E. C., J. J. Carr, P. L. Enright, E. A. Hoffman, A. R. Folsom, S. M. Kawut, R. A. Kronmal, D. J. Lederer, J. A. Lima, G. S. Lovasi, et al. 2016. Per cent emphysema is associated with respiratory and lung cancer mortality in the general population: a cohort study. Thorax 71: 624-632.
9. Houghton, A. M. 2013. Mechanistic links between COPD and lung cancer. Nat. Rev. Cancer 13: 233-245.
10. Hanahan, D., and R. A. Weinberg. 2011. Hallmarks of cancer: the next generation. Cell 144: 646-674.
11. Fan, Y., R. Mao, and J. Yang. 2013. NF-kB and STAT3 signaling pathways collaboratively link inflammation to cancer. Protein Cell 4: 176-185.
12. Umansky, V., C. Blattner, V. Fleming, X. Hu, C. Gebhardt, P. Altevogt, and J. Utikal. 2017. Myeloid-derived suppressor cells and tumor escape from immune surveillance. Semin. Immunopathol. 39: 295-305.
13. Wauters, E., W. Janssens, J. Vansteenkiste, H. Decaluwé, N. Heulens, B. Thienpont, H. Zhao, D. Smeets, X. Sagaert, J. Coolen, et al. 2015. DNA methylation profiling of non-small cell lung cancer reveals a COPD-driven immune-related signature. Thorax 70: 1113-1122.
14. Caetano, M. S., H. Zhang, A. M. Cumpian, L. Gong, N. Unver, E. J. Ostrin, S. Daliri, S. H. Chang, C. E. Ochoa, S. Hanash, et al. 2016. IL6 blockade reprograms the lung tumor microenvironment to limit the development and progression of K-ras-mutant lung cancer. Cancer Res. 76: 3189-3199.
15. Zaynagetdinov, R., T. P. Sherrill, L. A. Gleaves, P. Hunt, W. Han, A. G. McLoed, J. A. Saxon, H. Tanjore, P. M. Gulleman, L. R. Young, and T. S. Blackwell. 2016. Chronic NF-kB activation links COPD and lung cancer through generation of an immunosuppressive microenvironment in the lungs. Oncotarget 7: 5470-5482.
16. Schumacher, T. N., and R. D. Schreiber. 2015. Neoantigens in cancer immunotherapy. Science 348: 69-74.
17. Rizvi, N. A., M. D. Hellmann, A. Snyder, P. Kvistborg, V. Makarov, J. J. Havel, W. Lee, J. Yuan, P. Wong, T. S. Ho, et al. 2015. Cancer immunology. Mutational landscape determines sensitivity to PD-1 blockade in non-small cell lung cancer. Science 348: 124-128.
18. Gubin, M. M., X. Zhang, H. Schuster, E. Caron, J. P. Ward, T. Noguchi, Y. Ivanova, J. Hundal, C. D. Arthur, W.-J. Krebber, et al. 2014. Checkpoint blockade cancer immunotherapy targets tumour-specific mutant antigens. Nature 515: 577-581.
19. Tumeh, P. C., C. L. Harview, J. H. Yearley, I. P. Shintaku, E. J. Taylor, L. Robert, B. Chmielowski, M. Spasic, G. Henry, V. Ciobanu, et al. 2014. PD-1 blockade induces responses by inhibiting adaptive immune resistance. Nature 515: 568-571.
20. Goc, J., C. Germain, T. K. Vo-Bourgais, A. Lupo, C. Klein, S. Knockaert, L. de Chaisemartin, H. Ouakrim, E. Becht, M. Alifano, et al. 2014. Dendritic cells in tumor-associated tertiary lymphoid structures signal a Th1 cytotoxic immune contexture and license the positive prognostic value of infiltrating CD8+ T cells. Cancer Res. 74: 705-715.
21. Salmon, H., J. Idoyaga, A. Rahman, M. Leboeuf, R. Remark, S. Jordan, M. Casanova-Acebes, M. Khudoynazarova, J. Agudo, N. Tung, et al. 2016. Expansion and activation of CD103(+) dendritic cell progenitors at the tumor site enhances tumor responses to therapeutic PD-L1 and BRAF inhibition. Immunity 44: 924-938.
22. Tsoumakidou, M., S. Tousa, M. Semitekolou, P. Panagiotou, A. Panagiotou, I. Morianos, E. Litsiou, A. I. Trochoutsou, M. Konstantinou, K. Potaris, et al. 2014. Tolerogenic signaling by pulmonary CD1c+ dendritic cells induces regulatory T cells in patients with chronic obstructive pulmonary disease by IL-27/ IL-10/inducible costimulator ligand. J. Allergy Clin. Immunol. 134: 944-954.e8.
23. Brusselle, G. G., K. R. Bracke, T. Maes, A. I. D'hulst, K. B. Moerloose, G. F. Joos, and R. A. Pauwels. 2006. Murine models of COPD. Pulm. Pharmacol. Ther. 19: 155-165.
24. Stevenson, C. S., and M. A. Birrell. 2011. Moving towards a new generation of animal models for asthma and COPD with improved clinical relevance. Pharmacol. Ther. 130: 93-105.
25. D'hulst, A. I., K. Y. Vermaelen, G. G. Brusselle, G. F. Joos, and R. A. Pauwels. 2005. Time course of cigarette smoke-induced pulmonary inflammation in mice. Eur. Respir. J. 26: 204-213.
26. Dunnill, M. S. 1962. Quantitative methods in the study of pulmonary pathology. Thorax 17: 320-328.
27. Sarmiento, M., A. L. Glasebrook, and F. W. Fitch. 1980. IgG or IgM monoclonal antibodies reactive with different determinants on the molecular complex bearing Lyt 2 antigen block T cell-mediated cytolysis in the absence of complement. J. Immunol. 125: 2665-2672.
28. Robinson, M. D., D. J. McCarthy, and G. K. Smyth. 2010. edgeR: a bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 26: 139-140.
29. McCarthy, D. J., Y. Chen, and G. K. Smyth. 2012. Differential expression analysis of multifactor RNA-Seq experiments with respect to biological variation. Nucleic Acids Res. 40: 4288-4297.
30. Wickham, H. 2009. ggplot2: Elegant Graphics for Data Analysis. Springer-Verlag, New York.
31. Heberle, H., G. V. Meirelles, F. R. da Silva, G. P. Telles, and R. Minghim. 2015. InteractiVenn: a web-based tool for the analysis of sets through Venn diagrams. BMC Bioinformatics 16: 169.
32. Tripathi, S., M. O. Pohl, Y. Zhou, A. Rodriguez-Frandsen, G. Wang, D. A. Stein, H. M. Moulton, P. DeJesus, J. Che, L. C. Mulder, et al. 2015. Meta-and orthogonal integration of influenza "OMICs" data defines a role for UBR4 in virus budding. Cell Host Microbe 18: 723-735.
33. Gene Ontology Consortium. 2015. Gene ontology consortium: going forward. Nucleic Acids Res. 43: D1049-D1056.
34. Ashburner, M., C. A. Ball, J. A. Blake, D. Botstein, H. Butler, J. M. Cherry, A. P. Davis, K. Dolinski, S. S. Dwight, J. T. Eppig, et al. 2000. Gene ontology: tool for the unification of biology. Nat. Genet. 25: 25-29.
35. Fabregat, A., K. Sidiropoulos, P. Garapati, M. Gillespie, K. Hausmann, R. Haw, B. Jassal, S. Jupe, F. Korninger, S. McKay, et al. 2016. The reactome pathway knowledgebase. Nucleic Acids Res. 44: D481-D487.
36. Kellar, A., C. Egan, and D. Morris. 2015. Preclinical murine models for lung cancer: clinical trial applications. BioMed Res. Int. 2015: 621324.
37. Treffers, L. W., I. H. Hiemstra, T. W. Kuijpers, T. K. van den Berg, and H. L. Matlung. 2016. Neutrophils in cancer. Immunol. Rev. 273: 312-328.
38. Nielsen, S. R., and M. C. Schmid. 2017. Macrophages as key drivers of cancer progression and metastasis. Mediators Inflamm. 2017: 9624760.
39. Engblom, C., C. Pfirschke, and M. J. Pittet. 2016. The role of myeloid cells in cancer therapies. Nat. Rev. Cancer 16: 447-462.
40. Kiss, M., S. Van Gassen, K. Movahedi, Y. Saeys, and D. Laoui. 2018. Myeloid cell heterogeneity in cancer: not a single cell alike. Cell. Immunol. DOI: 10. 1016/j.cellimm.2018.02.008.
41. Marshall, E. A., K. W. Ng, S. H. Kung, E. M. Conway, V. D. Martinez, E. C. Halvorsen, D. A. Rowbotham, E. A. Vucic, A. W. Plumb, D. D. Becker-Santos, et al. 2016. Emerging roles of T helper 17 and regulatory T cells in lung cancer progression and metastasis. Mol. Cancer 15: 67.
42. Kedl, R. M., W. A. Rees, D. A. Hildeman, B. Schaefer, T. Mitchell, J. Kappler, and P. Marrack. 2000. T cells compete for access to antigen-bearing antigenpresenting cells. J. Exp. Med. 192: 1105-1113.
43. Denton, A. E., R. Wesselingh, S. Gras, C. Guillonneau, M. R. Olson, J. D. Mintern, W. Zeng, D. C. Jackson, J. Rossjohn, P. D. Hodgkin, et al. 2011. Affinity thresholds for naive CD8+ CTL activation by peptides and engineered influenza A viruses. J. Immunol. 187: 5733-5744.
44. Thompson, E. D., H. L. Enriquez, Y. X. Fu, and V. H. Engelhard. 2010. Tumor masses support naive T cell infiltration, activation, and differentiation into effectors. J. Exp. Med. 207: 1791-1804.
45. Paley, M. A., D. C. Kroy, P. M. Odorizzi, J. B. Johnnidis, D. V. Dolfi, B. E. Barnett, E. K. Bikoff, E. J. Robertson, G. M. Lauer, S. L. Reiner, and E. J. Wherry. 2012. Progenitor and terminal subsets of CD8+ T cells cooperate to contain chronic viral infection. Science 338: 1220-1225.
46. Tsoumakidou, M., A. V. Koutsopoulos, N. Tzanakis, K. Dambaki, E. Tzortzaki, S. Zakynthinos, P. K. Jeffery, and N. M. Siafakas. 2009. Decreased small airway and alveolar CD83+ dendritic cells in COPD. Chest 136: 726-733.
47. Robbins, C. S., F. Franco, M. Mouded, M. Cernadas, and S. D. Shapiro. 2008. Cigarette smoke exposure impairs dendritic cell maturation and T cell proliferation in thoracic lymph nodes of mice. J. Immunol. 180: 6623-6628.
48. Gibbons Johnson, R. M., and H. Dong. 2017. Functional expression of programmed death-ligand 1 (B7-H1) by immune cells and tumor cells. Front. Immunol. 8: 961.
49. Bhat, T. A., L. Panzica, S. G. Kalathil, and Y. Thanavala. 2015. Immune dysfunction in patients with chronic obstructive pulmonary disease. Ann. Am. Thorac. Soc. 12(Suppl. 2): S169-S175.
50. Kalathil, S. G., A. A. Lugade, V. Pradhan, A. Miller, G. I. Parameswaran, S. Sethi, and Y. Thanavala. 2014. T-regulatory cells and programmed death 1+ T cells contribute to effector T-cell dysfunction in patients with chronic obstructive pulmonary disease. Am. J. Respir. Crit. Care Med. 190: 40-50.
51. Biton, J., H. Ouakrim, A. Dechartres, M. Alifano, A. Mansuet-Lupo, H. Si, R. Halpin, T. Creasy, C. Bantsimba-Malanda, J. Arrondeau, et al. 2018. Impaired tumor-infiltrating T cells in patients with COPD impacts lung cancer response to PD-1 blockade. Am. J. Respir. Crit. Care Med. DOI: 10.1164/rccm.201706-1110OC.
52. Mark, N. M., J. Kargl, S. E. Busch, G. H. Y. Yang, H. E. Metz, H. Zhang, J. J. Hubbard, S. N. J. Pipavath, D. K. Madtes, and A. M. Houghton. 2018. Chronic obstructive pulmonary disease alters immune cell composition and immune checkpoint inhibitor efficacy in non-small cell lung cancer. Am. J. Respir. Crit. Care Med. 197: 325-336.
53. Barnes, P. J. 2016. Inflammatory mechanisms in patients with chronic obstructive pulmonary disease. J. Allergy Clin. Immunol. 138: 16-27.
54. Stockley, R. A. 1999. Neutrophils and protease/antiprotease imbalance. Am. J. Respir. Crit. Care Med. 160(suppl. 1): S49-S52.
55. Domej, W., K. Oettl, and W. Renner. 2014. Oxidative stress and free radicals in COPD-implications and relevance for treatment. Int. J. Chron. Obstruct. Pulmon. Dis. 9: 1207-1224.
56. Johnson, S. R. 2016. Untangling the protease web in COPD: metalloproteinases in the silent zone. Thorax 71: 105-106.
57. Barnes, P. J. 2014. Cellular and molecular mechanisms of chronic obstructive pulmonary disease. Clin. Chest Med. 35: 71-86.
58. Wang, J., Q. Li, J. Xie, and Y. Xu. 2015. Cigarette smoke inhibits BAFF expression and mucosal immunoglobulin A responses in the lung during influenza virus infection. Respir. Res. 16: 37.
59. Lugade, A. A., P. N. Bogner, T. H. Thatcher, P. J. Sime, R. P. Phipps, and Y. Thanavala. 2014. Cigarette smoke exposure exacerbates lung inflammation and compromises immunity to bacterial infection. J. Immunol. 192: 5226-5235.
60. Gotts, J. E., L. F. Chun, J. Abbott, X. Fang, N. Takasaki, S. L. Nishimura, M. L. Springer, S. F. Schick, C. S. Calfee, and M. A. Matthay. 2018. Cigarette smoke exposure worsens acute lung injury in antibiotic-treated bacterial pneumonia in mice. Am. J. Physiol. Lung Cell. Mol. Physiol. 315: L25-L40.
61. Shen, P., M. C. Morissette, G. Vanderstocken, Y. Gao, M. Hassan, A. Roos, D. Thayaparan, M. Merlano, M. G. Dorrington, J. K. Nikota, et al. 2016. Cigarette smoke attenuates the nasal host response to Streptococcus pneumoniae and predisposes to invasive pneumococcal disease in mice. Infect. Immun. 84: 1536-1547.
62. Lanckacker, E. A., K. G. Tournoy, H. Hammad, G. Holtappels, B. N. Lambrecht, G. F. Joos, and T. Maes. 2013. Short cigarette smoke exposure facilitates sensitisation and asthma development in mice. Eur. Respir. J. 41: 1189-1199.
63. Lo Sasso, G., W. K. Schlage, S. Boué, E. Veljkovic, M. C. Peitsch, and J. Hoeng. 2016. The Apoe(-/-) mouse model: a suitable model to study cardiovascular and respiratory diseases in the context of cigarette smoke exposure and harm reduction. J. Transl. Med. 14: 146.
64. Coggins, C. R. 2010. A further review of inhalation studies with cigarette smoke and lung cancer in experimental animals, including transgenic mice. Inhal. Toxicol. 22: 974-983.
65. Ambrogio, C., F. J. Carmona, A. Vidal, M. Falcone, P. Nieto, O. A. Romero, S. Puertas, M. Vizoso, E. Nadal, T. Poggio, et al. 2014. Modeling lung cancer evolution and preclinical response by orthotopic mouse allografts. Cancer Res. 74: 5978-5988.
66. Witschi, H., I. Espiritu, J. L. Peake, K. Wu, R. R. Maronpot, and K. E. Pinkerton. 1997. The carcinogenicity of environmental tobacco smoke. Carcinogenesis 18: 575-586.
67. Kerdidani, D., K. Glynos, S. Magkouta, G. Kazamias, V. Karavana, C. Roussos, D. Rontogianni, I. Kalomenidis, S. Zakynthinos, and M. Tsoumakidou. 2015. Deciphering the link between COPD and lung cancer: cigarette smoke-induced immunosuppression at the crossroads between cancer immune surveillance and escape. Eur. Respir. J. 46: PA1898.
68. Tsoumakidou, M., I. K. Demedts, G. G. Brusselle, and P. K. Jeffery. 2008. Dendritic cells in chronic obstructive pulmonary disease: new players in an old game. Am. J. Respir. Crit. Care Med. 177: 1180-1186.
69. Wherry, E. J., and M. Kurachi. 2015. Molecular and cellular insights into T cell exhaustion. Nat. Rev. Immunol. 15: 486-499.
70. Tassi, E., G. Grazia, C. Vegetti, I. Bersani, G. Bertolini, A. Molla, P. Baldassari, F. Andriani, L. Roz, G. Sozzi, et al. 2017. Early effector T lymphocytes coexpress multiple inhibitory receptors in primary non-small cell lung cancer. Cancer Res. 77: 851-861.
71. Schietinger, A., M. Philip, V. E. Krisnawan, E. Y. Chiu, J. J. Delrow, R. S. Basom, P. Lauer, D. G. Brockstedt, S. E. Knoblaugh, G. J. Hämmerling, et al. 2016. Tumorspecific T cell dysfunction is a dynamic antigen-driven differentiation program initiated early during tumorigenesis. Immunity 45: 389-401.
72. Pauken, K. E., and E. J. Wherry. 2015. Overcoming T cell exhaustion in infection and cancer. Trends Immunol. 36: 265-276.
73. Demoor, T., K. R. Bracke, G. F. Joos, and G. G. Brusselle. 2010. Increased Tregulatory cells in lungs and draining lymph nodes in a murine model of COPD. Eur. Respir. J. 35: 688-689.
74. Sade-Feldman, M., J. Kanterman, E. Ish-Shalom, M. Elnekave, E. Horwitz, and M. Baniyash. 2013. Tumor necrosis factor-A blocks differentiation and enhances suppressive activity of immature myeloid cells during chronic inflammation. Immunity 38: 541-554.
75. Han, S., A. Asoyan, H. Rabenstein, N. Nakano, and R. Obst. 2010. Role of antigen persistence and dose for CD4+ T-cell exhaustion and recovery. Proc. Natl. Acad. Sci. USA 107: 20453-20458.
76. Motz, G. T., B. L. Eppert, S. C. Wesselkamper, J. L. Flury, and M. T. Borchers. 2010. Chronic cigarette smoke exposure generates pathogenic T cells capable of driving COPD-like disease in Rag2-/-mice. Am. J. Respir. Crit. Care Med. 181: 1223-1233.
77. Maeno, T., A. M. Houghton, P. A. Quintero, S. Grumelli, C. A. Owen, and S. D. Shapiro. 2007. CD8+ T cells are required for inflammation and destruction in cigarette smoke-induced emphysema in mice. J. Immunol. 178: 8090-8096.
78. Podolin, P. L., J. P. Foley, D. C. Carpenter, B. J. Bolognese, G. A. Logan, E. Long, III, O. J. Harrison, and P. T. Walsh. 2013. T cell depletion protects against alveolar destruction due to chronic cigarette smoke exposure in mice. Am. J. Physiol. Lung Cell. Mol. Physiol. 304: L312-L323.
79. Borghaei, H., L. Paz-Ares, L. Horn, D. R. Spigel, M. Steins, N. E. Ready, L. Q. Chow, E. E. Vokes, E. Felip, E. Holgado, et al. 2015. Nivolumab versus docetaxel in advanced nonsquamous non-small-cell lung cancer. N. Engl. J. Med. 373: 1627-1639.
80. Rittmeyer, A., F. Barlesi, D. Waterkamp, K. Park, F. Ciardiello, J. von Pawel, S. M. Gadgeel, T. Hida, D. M. Kowalski, M. C. Dols, et al; OAK Study Group. 2017. Atezolizumab versus docetaxel in patients with previously treated nonsmall-cell lung cancer (OAK): a phase 3, open-label, multicentre randomised controlled trial. Lancet 389: 255-265.