摘要
弥漫性大B细胞淋巴瘤(DLBCL)是成人中最常见的B型非霍奇金淋巴瘤。 引入利妥昔单抗治疗DLBCL后,当前的一线治疗是R-CHOP方案。 该方案极大地改善了患者的预后,但是,复发或难治性病例屡见不鲜,主要是由于对利妥昔单抗的耐药性。 尽管已经进行了大量实验来研究利妥昔单抗的耐药性,但是其exac机制和解决方案仍不清楚。 这篇综述主要探讨了对利妥昔单抗耐药的可能机制和目前对DLBCL利妥昔单抗耐药的新有效治疗方法。
关键词: 弥漫性大B细胞淋巴瘤,CD20,利妥昔单抗耐药,生物标志物,非霍奇金淋巴瘤,预后。
[1]
Martelli, M.; Ferreri, A.J.; Agostinelli, C.; Di Rocco, A.; Pfreundschuh, M.; Pileri, S.A. Diffuse large B-cell lymphoma. Crit. Rev. Oncol. Hematol., 2013, 87(2), 146-171.
[2]
Schneider, C.; Pasqualucci, L.; Dalla-Favera, R. Molecular pathogenesis of diffuse large B-cell lymphoma. Semin. Diagn. Pathol., 2011, 28(2), 167-177.
[3]
Shaffer, A.L., III; Young, R.M.; Staudt, L.M. Pathogenesis of human B cell lymphomas. Annu. Rev. Immunol., 2012, 30, 565-610.
[4]
Nastoupil, L.J.; Rose, A.C.; Flowers, C.R. Diffuse large B-cell lymphoma: current treatment approaches. Oncology (Williston Park), 2012, 26(5), 488-495.
[5]
Tedder, T.F.; Disteche, C.M.; Louie, E.; Adler, D.A.; Croce, C.M.; Schlossman, S.F.; Saito, H. The gene that encodes the human CD20 (B1) differentiation antigen is located on chromosome 11 near the t(11;14)(q13;q32) translocation site. J. Immunol., 1989, 142(7), 2555-2559.
[6]
Henry, C.; Deschamps, M.; Rohrlich, P.S.; Pallandre, J.R.; Rémy-Martin, J.P.; Callanan, M.; Traverse-Glehen, A.; GrandClément, C.; Garnache-Ottou, F.; Gressin, R.; Deconinck, E.; Salles, G.; Robinet, E.; Tiberghien, P.; Borg, C.; Ferrand, C. Identification of an alternative CD20 transcript variant in B-cell malignancies coding for a novel protein associated to rituximab resistance. Blood, 2010, 115(12), 2420-2429.
[7]
Maeshima, A.M.; Taniguchi, H.; Fukuhara, S.; Morikawa, N.; Munakata, W.; Maruyama, D.; Kim, S.W.; Watanabe, T.; Kobayashi, Y.; Tobinai, K.; Tsuda, H. Follow-up data of 10 patients with B-cell non-Hodgkin lymphoma with a CD20-negative phenotypic change after rituximab-containing therapy. Am. J. Surg. Pathol., 2013, 37(4), 563-570.
[8]
Matsuda, I.; Hirota, S. Bone marrow infiltration of CD20-negative follicular lymphoma after rituximab therapy: A histological mimicker of hematogones and B-cell acute lymphoblastic leukemia/lymphoma. Int. J. Clin. Exp. Pathol., 2015, 8(8), 9737-9741.
[9]
Tsutsumi, Y.; Ohigashi, H.; Ito, S.; Shiratori, S.; Teshima, T. 5-Azacytidine partially restores CD20 expression in follicular lymphoma that lost CD20 expression after rituximab treatment: a case report. J. Med. Case Reports, 2016, 10, 27.
[10]
Small, G.W.; McLeod, H.L.; Richards, K.L. Analysis of innate and acquired resistance to anti-CD20 antibodies in malignant and nonmalignant B cells. Peer J, 2013. 1e31
[11]
Terui, Y.; Mishima, Y.; Sugimura, N.; Kojima, K.; Sakurai, T.; Mishima, Y.; Kuniyoshi, R.; Taniyama, A.; Yokoyama, M.; Sakajiri, S.; Takeuchi, K.; Watanabe, C.; Takahashi, S.; Ito, Y.; Hatake, K. Identification of CD20 C-terminal deletion mutations associated with loss of CD20 expression in non-Hodgkin’s lymphoma. Clin. Cancer Res., 2009, 15(7), 2523-2530.
[12]
Mishima, Y.; Terui, Y.; Takeuchi, K.; Matsumoto-Mishima, Y.; Matsusaka, S.; Utsubo-Kuniyoshi, R.; Hatake, K. The identification of irreversible rituximab-resistant lymphoma caused by CD20 gene mutations. Blood Cancer J., 2011, 1(4) e15
[13]
Johnson, N.A.; Leach, S.; Woolcock, B.; deLeeuw, R.J.; Bashashati, A.; Sehn, L.H.; Connors, J.M.; Chhanabhai, M.; Brooks-Wilson, A.; Gascoyne, R.D. CD20 mutations involving the rituximab epitope are rare in diffuse large B-cell lymphomas and are not a significant cause of R-CHOP failure. Haematologica, 2009, 94(3), 423-427.
[14]
Sar, A.; Perizzolo, M.; Stewart, D.; Mansoor, A.; Difrancesco, L.M.; Demetrick, D.J. Mutation or polymorphism of the CD20 gene is not associated with the response to R-CHOP in diffuse large B cell lymphoma patients. Leuk. Res., 2009, 33(6), 792-797.
[15]
Davis, T.A.; Czerwinski, D.K.; Levy, R. Therapy of B-cell lymphoma with anti-CD20 antibodies can result in the loss of CD20 antigen expression. Clin. Cancer Res., 1999, 5(3), 611-615.
[16]
Pedersen, A.E.; Jungersen, M.B.; Pedersen, C.D. Monocytes mediate shaving of B-cell-bound anti-CD20 antibodies. Immunology, 2011, 133(2), 239-245.
[17]
Czuczman, M.S.; Olejniczak, S.; Gowda, A.; Kotowski, A.; Binder, A.; Kaur, H.; Knight, J.; Starostik, P.; Deans, J.; Hernandez-Ilizaliturri, F.J. Acquirement of rituximab resistance in lymphoma cell lines is associated with both global CD20 gene and protein down-regulation regulated at the pretranscriptional and posttranscriptional levels. Clin. Cancer Res., 2008, 14(5), 1561-1570.
[18]
Ziepert, M.; Hasenclever, D.; Kuhnt, E.; Glass, B.; Schmitz, N.; Pfreundschuh, M.; Loeffler, M. Standard International prognostic index remains a valid predictor of outcome for patients with aggressive CD20+ B-cell lymphoma in the rituximab era. J. Clin. Oncol., 2010, 28(14), 2373-2380.
[19]
Hatjiharissi, E.; Xu, L.; Santos, D.D.; Hunter, Z.R.; Ciccarelli, B.T.; Verselis, S.; Modica, M.; Cao, Y.; Manning, R.J.; Leleu, X.; Dimmock, E.A.; Kortsaris, A.; Mitsiades, C.; Anderson, K.C.; Fox, E.A.; Treon, S.P. Increased natural killer cell expression of CD16, augmented binding and ADCC activity to rituximab among individuals expressing the FcgammaRIIIa-158 V/V and V/F polymorphism. Blood, 2007, 110(7), 2561-2564.
[20]
Danielou-Lazareth, A.; Henry, G.; Geromin, D.; Khaznadar, Z.; Briere, J.; Tamouza, R.; Cayuela, J.M.; Thieblemont, C.; Toubert, A.; Dulphy, N. At diagnosis, diffuse large B-cell lymphoma patients show impaired rituximab-mediated NK-cell cytotoxicity. Eur. J. Immunol., 2013, 43(5), 1383-1388.
[21]
Weng, W.K.; Levy, R. Two immunoglobulin G fragment C receptor polymorphisms independently predict response to rituximab in patients with follicular lymphoma. J. Clin. Oncol., 2003, 21(21), 3940-3947.
[22]
Vega, M.I.; Huerta-Yepez, S.; Martinez-Paniagua, M.; Martinez-Miguel, B.; Hernandez-Pando, R.; González-Bonilla, C.R.; Chinn, P.; Hanna, N.; Hariharan, K.; Jazirehi, A.R.; Bonavida, B. Rituximab-mediated cell signaling and chemo/immuno-sensitization of drug-resistant B-NHL is independent of its Fc functions. Clin. Cancer Res., 2009, 15(21), 6582-6594.
[23]
Rezvani, A.R.; Maloney, D.G. Rituximab resistance. Best Pract. Res. Clin. Haematol., 2011, 24(2), 203-216.
[24]
Takei, K.; Yamazaki, T.; Sawada, U.; Ishizuka, H.; Aizawa, S. Analysis of changes in CD20, CD55, and CD59 expression on established rituximab-resistant B-lymphoma cell lines. Leuk. Res., 2006, 30(5), 625-631.
[25]
Ziller, F.; Macor, P.; Bulla, R.; Sblattero, D.; Marzari, R.; Tedesco, F. Controlling complement resistance in cancer by using human monoclonal antibodies that neutralize complement-regulatory proteins CD55 and CD59. Eur. J. Immunol., 2005, 35(7), 2175-2183.
[26]
Macor, P.; Tripodo, C.; Zorzet, S.; Piovan, E.; Bossi, F.; Marzari, R.; Amadori, A.; Tedesco, F. In vivo targeting of human neutralizing antibodies against CD55 and CD59 to lymphoma cells increases the antitumor activity of rituximab. Cancer Res., 2007, 67(21), 10556-10563.
[27]
Hu, W.; Ge, X.; You, T.; Xu, T.; Zhang, J.; Wu, G. Human CD59 inhibitor sensitizes rituximab-resistant lymphoma cells to complement-mediated cytolysis. Cancer Res., 2011, 71(6), 2298-2307.
[28]
Mamidi, S.; Höne, S.; Teufel, C.; Sellner, L.; Zenz, T.; Kirschfink, M. Neutralization of membrane complement regulators improves complement-dependent effector functions of therapeutic anticancer antibodies targeting leukemic cells. OncoImmunology, 2015, 4(3) e979688
[29]
Makou, E.; Herbert, A.P.; Barlow, P.N. Functional anatomy of complement factor H. Biochemistry, 2013, 52(23), 3949-3962.
[30]
Jazirehi, A.R.; Vega, M.I.; Bonavida, B. Development of rituximab-resistant lymphoma clones with altered cell signaling and cross-resistance to chemotherapy. Cancer Res., 2007, 67(3), 1270-1281.
[31]
Gomez-Gelvez, J.C.; Salama, M.E.; Perkins, S.L.; Leavitt, M.; Inamdar, K.V. Prognostic impact of tumor microenvironment in diffuse large B-cell lymphoma uniformly treated with R-CHOP chemotherapy. Am. J. Clin. Pathol., 2016, 145(4), 514-523.
[32]
Fridman, W.H.; Pagès, F.; Sautès-Fridman, C.; Galon, J. The immune contexture in human tumours: impact on clinical outcome. Nat. Rev. Cancer, 2012, 12(4), 298-306.
[33]
Zhong, W.; Xu, X.; Zhu, Z.; Du, Q.; Du, H.; Yang, L.; Ling, Y.; Xiong, H.; Li, Q. Increased expression of IRF8 in tumor cells inhibits the generation of Th17 cells and predicts unfavorable survival of diffuse large B cell lymphoma patients. Oncotarget, 2017, 8(30), 49757-49772.
[34]
Lv, X.; Feng, L.; Ge, X.; Lu, K.; Wang, X. Interleukin-9 promotes cell survival and drug resistance in diffuse large B-cell lymphoma. J. Exp. Clin. Cancer Res., 2016, 35(1), 106.
[35]
Lykken, J.M.; Horikawa, M.; Minard-Colin, V.; Kamata, M.; Miyagaki, T.; Poe, J.C.; Tedder, T.F. Galectin-1 drives lymphoma CD20 immunotherapy resistance: validation of a preclinical system to identify resistance mechanisms. Blood, 2016, 127(15), 1886-1895.
[36]
Alas, S.; Emmanouilides, C.; Bonavida, B. Inhibition of interleukin 10 by rituximab results in down-regulation of bcl-2 and sensitization of B-cell non-Hodgkin’s lymphoma to apoptosis. Clin. Cancer Res., 2001, 7(3), 709-723.
[37]
Challa-Malladi, M.; Lieu, Y.K.; Califano, O.; Holmes, A.B.; Bhagat, G.; Murty, V.V.; Dominguez-Sola, D.; Pasqualucci, L.; Dalla-Favera, R. Combined genetic inactivation of β2-Microglobulin and CD58 reveals frequent escape from immune recognition in diffuse large B cell lymphoma. Cancer Cell, 2011, 20(6), 728-740.
[38]
Bittenbring, J.T.; Neumann, F.; Altmann, B.; Achenbach, M.; Reichrath, J.; Ziepert, M.; Geisel, J.; Regitz, E.; Held, G.; Pfreundschuh, M. Vitamin D deficiency impairs rituximab-mediated cellular cytotoxicity and outcome of patients with diffuse large B-cell lymphoma treated with but not without rituximab. J. Clin. Oncol., 2014, 32(29), 3242-3248.
[39]
Roschewski, M.; Staudt, L.M.; Wilson, W.H. Diffuse large B-cell lymphoma-treatment approaches in the molecular era. Nat. Rev. Clin. Oncol., 2014, 11(1), 12-23.
[40]
Bai, D.; Ueno, L.; Vogt, P.K. Akt-mediated regulation of NF kappaB and the essentialness of NF kappaB for the oncogenicity of PI3K and Akt. Int. J. Cancer, 2009, 125, 2863-2870.
[41]
Fan, Y.; Mao, R.; Yang, J. NF-κB and STAT3 signaling pathways collaboratively link inflammation to cancer. Protein Cell, 2013, 4(3), 176-185.
[42]
Ma, Y.; Zhang, P.; Gao, Y.; Fan, H.; Zhang, M.; Wu, J. Evaluation of AKT phosphorylation and PTEN loss and their correlation with the resistance of rituximab in DLBCL. Int. J. Clin. Exp. Pathol., 2015, 8(11), 14875-14884.
[43]
Zhong, W.; Xu, X.; Zhu, Z.; Yang, L.; Du, H.; Xia, Z.; Yuan, Z.; Xiong, H.; Du, Q.; Wei, Y.; Li, Q. Increased interleukin-17A levels promote rituximab resistance by suppressing p53 expression and predict an unfavorable prognosis in patients with diffuse large B cell lymphoma. Int. J. Oncol., 2018, 4299.
[44]
Sarkozy, C.; Traverse-Glehen, A.; Coiffier, B. Double-hit and double-protein-expression lymphomas: Aggressive and refractory lymphomas. Lancet Oncol., 2015, 16(15), e555-e567.
[45]
Burotto, M.; Berkovits, A.; Dunleavy, K. Double hit lymphoma: from biology to therapeutic implications. Expert Rev. Hematol., 2016, 9(7), 669-678.
[46]
Juskevicius, D.; Jucker, D.; Klingbiel, D.; Mamot, C.; Dirnhofer, S.; Tzankov, A. Mutations of CREBBP and SOCS1 are independent prognostic factors in diffuse large B cell lymphoma: mutational analysis of the SAKK 38/07 prospective clinical trial cohort. J. Hematol. Oncol., 2017, 10(1), 70.
[47]
Knudsen, S.; Hother, C.; Grønbæk, K.; Jensen, T.; Hansen, A.; Mazin, W.; Dahlgaard, J.; Møller, M.B.; Ralfkiær, E. Brown, Pde. N. Development and blind clinical validation of a microRNA based predictor of response to treatment with R-CHO(E)P in DLBCL. PLoS One, 2015, 10(2)e0115538
[48]
Song, G.; Song, G.; Ni, H.; Gu, L.; Liu, H.; Chen, B.; He, B.; Pan, Y.; Wang, S.; Cho, W.C. Deregulated expression of miR-224 and its target gene: CD59 predicts outcome of diffuse large B-cell lymphoma patients treated with R-CHOP. Curr. Cancer Drug Targets, 2014, 14(7), 659-670.
[49]
Iqbal, J.; Shen, Y.; Huang, X.; Liu, Y.; Wake, L. Global microRNA expression profiling uncovers molecular markers for classification and prognosis in aggressive B-cell lymphoma. Blood, 2015, 125, 1137-1145.
[50]
Gu, J.J.; Hernandez-Ilizaliturri, F.J.; Mavis, C.; Czuczman, N.M.; Deeb, G.; Gibbs, J.; Skitzki, J.J.; Patil, R.; Czuczman, M.S. MLN2238, a proteasome inhibitor, induces caspase-dependent cell death, cell cycle arrest, and potentiates the cytotoxic activity of chemotherapy agents in rituximab-chemotherapy-sensitive or rituximab-chemotherapy-resistant B-cell lymphoma preclinical models. Anticancer Drugs, 2013, 24(10), 1030-1038.
[51]
Olejniczak, S.H.; Blickwedehl, J.; Belicha-Villanueva, A.; Bangia, N.; Riaz, W.; Mavis, C.; Clements, J.L.; Gibbs, J.; Hernandez-Ilizaliturri, F.J.; Czuczman, M.S. Distinct molecular mechanisms responsible for bortezomib-induced death of therapy-resistant versus -sensitive B-NHL cells. Blood, 2010, 116(25), 5605-5614.
[52]
Barr, P.; Fisher, R.; Friedberg, J. The role of bortezomib in the treatment of lymphoma. Cancer Invest., 2007, 25(8), 766-775.
[53]
Bil, J.; Winiarska, M.; Nowis, D.; Bojarczuk, K.; Dabrowska-Iwanicka, A.; Basak, G.W.; Sułek, K.; Jakobisiak, M.; Golab, J. Bortezomib modulates surface CD20 in B-cell malignancies and affects rituximab-mediated complement-dependent cytotoxicity. Blood, 2010, 115(18), 3745-3755.
[54]
Advani, R.H.; Buggy, J.J.; Sharman, J.P.; Smith, S.M.; Boyd, T.E.; Grant, B.; Kolibaba, K.S.; Furman, R.R.; Rodriguez, S.; Chang, B.Y.; Sukbuntherng, J.; Izumi, R.; Hamdy, A.; Hedrick, E.; Fowler, N.H. Bruton tyrosine kinase inhibitor ibrutinib (PCI-32765) has significant activity in patients with relapsed/refractory B-cell malignancies. J. Clin. Oncol., 2013, 31(1), 88-94.
[55]
Johnston, P.B.; LaPlant, B.; McPhail, E.; Habermann, T.M.; Inwards, D.J.; Micallef, I.N. Everolimus combined with R-CHOP-21 for new, untreated, diffuse large B-cell lymphoma (NCCTG 1085 [Alliance]): Safety and efficacy results of a phase 1 and feasibility trial. Lancet Haematol., 2016, 3(7), e309-e316.
[56]
Galustian, C.; Meyer, B.; Labarthe, M.C.; Dredge, K.; Klaschka, D.; Henry, J.; Todryk, S.; Chen, R.; Muller, G.; Stirling, D.; Schafer, P.; Bartlett, J.B.; Dalgleish, A.G. The anti-cancer agents lenalidomide and pomalidomide inhibit the proliferation and function of T regulatory cells. Cancer Immunol. Immunother., 2009, 58(7), 1033-1045.
[57]
Nowakowski, G.S.; LaPlant, B.; Macon, W.R.; Reeder, C.B.; Foran, J.M.; Nelson, G.D.; Thompson, C.A.; Rivera, C.E.; Inwards, D.J.; Micallef, I.N.; Johnston, P.B.; Porrata, L.F.; Ansell, S.M.; Gascoyne, R.D.; Habermann, T.M.; Witzig, T.E. Lenalidomide combined with R-CHOP overcomes negative prognostic impact of non-germinal center B-cell phenotype in newly diagnosed diffuse large B-Cell lymphoma: a phase II study. J. Clin. Oncol., 2015, 33(3), 251-257.
[58]
Chiappella, A.; Tucci, A.; Castellino, A.; Pavone, V.; Baldi, I.; Carella, A.M.; Orsucci, L.; Zanni, M.; Salvi, F.; Liberati, A.M.; Gaidano, G.; Bottelli, C.; Rossini, B.; Perticone, S.; De Masi, P.; Ladetto, M.; Ciccone, G.; Palumbo, A.; Rossi, G.; Vitolo, U. Lenalidomide plus cyclophosphamide, doxorubicin, vincristine, prednisone and rituximab is safe and effective in untreated, elderly patients with diffuse large B-cell lymphoma: A phase I study by the Fondazione Italiana Linfomi. Haematologica, 2013, 98(11), 1732-1738.
[59]
Kohrt, H.E.; Houot, R.; Goldstein, M.J.; Weiskopf, K.; Alizadeh, A.A.; Brody, J.; Müller, A.; Pachynski, R.; Czerwinski, D.; Coutre, S.; Chao, M.P.; Chen, L.; Tedder, T.F.; Levy, R. CD137 stimulation enhances the antilymphoma activity of anti-CD20 antibodies. Blood, 2011, 117(8), 2423-2432.
[60]
Kohrt, H.E.; Thielens, A.; Marabelle, A.; Sagiv-Barfi, I.; Sola, C.; Chanuc, F.; Fuseri, N.; Bonnafous, C.; Czerwinski, D.; Rajapaksa, A.; Waller, E.; Ugolini, S.; Vivier, E.; Romagné, F.; Levy, R.; Bléry, M.; André, P. Anti-KIR antibody enhancement of anti-lymphoma activity of natural killer cells as monotherapy and in combination with anti-CD20 antibodies. Blood, 2014, 123(5), 678-686.
[61]
Deguine, J.; Breart, B.; Lemaître, F.; Bousso, P. Cutting edge: tumor-targeting antibodies enhance NKG2D-mediated NK cell cytotoxicity by stabilizing NK cell-tumor cell interactions. J. Immunol., 2012, 189(12), 5493-5497.
[62]
Bhatt, S.; Parvin, S.; Zhang, Y.; Cho, H.M.; Kunkalla, K.; Vega, F.; Timmerman, J.M.; Shin, S.U.; Rosenblatt, J.D.; Lossos, I.S. Anti-CD20-Interleukin-21 fusokine targets malignant B-cells via direct apoptosis and NK-cell dependent cytotoxicity. Blood, 2017, 129(16), 2246-2256.
[63]
Jurczak, W.; Zinzani, P.L.; Gaidano, G.; Goy, A.; Provencio, M.; Nagy, Z.; Robak, T.; Maddocks, K.; Buske, C.; Ambarkhane, S.; Winderlich, M.; Dirnberger-Hertweck, M.; Korolkiewicz, R.; Blum, K.A. Phase IIa study of the CD19 antibody MOR208 in patients with relapsed or refractory B-cell non-Hodgkin’s lymphoma. Ann. Oncol., 2018, 29(5), 1266-1272.
[64]
Schuster, S.J.; Svoboda, J.; Chong, E.A.; Nasta, S.D.; Mato, A.R.; Anak, Ö.; Brogdon, J.L. Chimeric antigen receptor T cells in refractory B-cell lymphomas. N. Engl. J. Med., 2017, 377(26), 2545-2554.
[65]
Lesokhin, A.M.; Ansell, S.M.; Armand, P.; Scott, E.C.; Halwani, A.; Gutierrez, M.; Millenson, M.M.; Cohen, A.D.; Schuster, S.J.; Lebovic, D.; Dhodapkar, M. Nivolumab in patients with relapsed or refractory hematologic malignancy: Preliminary results of a phase 1b study. J. Clin. Oncol., 2016, 34(23), 2698-2704.
[66]
Hayashi, K.; Nagasaki, E.; Kan, S.; Ito, M.; Kamata, Y.; Homma, S.; Aiba, K. Gemcitabine enhances rituximab-mediated complement-dependent cytotoxicity to B cell lymphoma by CD20 upregulation. Cancer Sci., 2016, 107(5), 682-689.
[67]
Vega, G.G.; Franco-Cea, L.A.; Huerta-Yepez, S.; Mayani, H.; Morrison, S.L.; Bonavida, B.; Vega, M.I. Overcoming rituximab drug-resistance by the genetically engineered anti-CD20-hIFN-α fusion protein: Direct cytotoxicity and synergy with chemotherapy. Int. J. Oncol., 2015, 47(5), 1735-1748.
[68]
Richter, M.; Yumul, R.; Saydaminova, K.; Wang, H.; Gough, M.; Baldessari, A.; Cattaneo, R.; Lee, F.; Wang, C.H.; Jang, H.; Astier, A.; Gopal, A.; Carter, D.; Lieber, A. Preclinical safety, pharmacokinetics, pharmacodynamics, and biodistribution studies with Ad35K++ protein: a novel rituximab cotherapeutic. Mol. Ther. Methods Clin. Dev., 2016, 5, 16013.