Thromb Haemost 2018; 118(10): 1752-1764
DOI: 10.1055/s-0038-1669921
Cellular Haemostasis and Platelets
Georg Thieme Verlag KG Stuttgart · New York

Proteasome Inhibition with Bortezomib Induces Apoptosis of Long-Lived Plasma Cells in Steroid-Resistant or Relapsed Immune Thrombocytopaenia

Guosheng Li*
1   Department of Haematology, Qilu Hospital, Shandong University, Jinan, China
,
Shuang Wang*
1   Department of Haematology, Qilu Hospital, Shandong University, Jinan, China
,
Na Li
2   Key Laboratory of Cardiovascular Remodelling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Jinan, China
,
Yang Liu
1   Department of Haematology, Qilu Hospital, Shandong University, Jinan, China
,
Qi Feng
1   Department of Haematology, Qilu Hospital, Shandong University, Jinan, China
,
Xinyi Zuo
1   Department of Haematology, Qilu Hospital, Shandong University, Jinan, China
,
Xin Li
1   Department of Haematology, Qilu Hospital, Shandong University, Jinan, China
,
Yu Hou
1   Department of Haematology, Qilu Hospital, Shandong University, Jinan, China
,
Linlin Shao
1   Department of Haematology, Qilu Hospital, Shandong University, Jinan, China
,
Chunhong Ma
3   Department of Immunology, Shandong University School of Medicine, Shandong University, Jinan, China
,
Chengjiang Gao
3   Department of Immunology, Shandong University School of Medicine, Shandong University, Jinan, China
,
Ming Hou
2   Key Laboratory of Cardiovascular Remodelling and Function Research, Chinese Ministry of Education and Chinese Ministry of Health, Jinan, China
4   Shandong Provincial Key Laboratory of Immunohematology, Qilu Hospital, Shandong University, Jinan, China
,
Jun Peng
1   Department of Haematology, Qilu Hospital, Shandong University, Jinan, China
› Author Affiliations
Funding This work was supported by grants from the Major Research plan of the National Natural Science Foundation of China (91442204), National Natural Science Foundation for Distinguished Young Scholars of China (81125002), National Natural Science Foundation of China (81370623, 81770133, 81770114, 81470284 and 81500094), Natural Science Foundation of Shandong Province (ZR2017PH022, ZR2017PH041, ZR2016HP01) and Tai Shan Scholar Foundation.
Further Information

Publication History

14 January 2018

06 August 2018

Publication Date:
20 September 2018 (online)

Abstract

Primary immune thrombocytopaenia (ITP) is the most common haemorrhagic disease. Although most patients respond initially to mainstream therapies, such as corticosteroids, immunosuppressants or rituximab, a large proportion of patients fail to respond or relapse. These treatments only affect B lymphocytes or short-lived plasma cells, but not already existing long-lived plasma cells (LLPCs) which persistently secrete antibodies. We hypothesized that LLPCs may play a role in the corticosteroid-resistant or relapsed ITP patients, and bortezomib, a proteasome inhibitor, may act on plasma cells and offer a therapeutic effect. Although a significant difference in the proportion of CD19CD38hiCD138+ total LLPCs was not observed by flow cytometry, a glycoprotein (GP) IIb/IIIa-specific enzyme-linked immunosorbent spot (ELISpot) assay of sorted CD19CD138+ LLPCs confirmed the existence of anti-platelet antibody-secreting LLPCs in ITP patients in contrast to healthy controls. Moreover, the LLPCs could be eliminated in the presence of bortezomib by ELISpot assay, which was also confirmed by flow cytometry. Accordingly, a modified monoclonal antibody immobilization of platelet antigen assay of sorted CD19CD138+ LLPCs revealed that the concentration of anti-platelet antibodies decreased remarkably when cultured with 0.25 ng/mL bortezomib for 5 days. The apoptosis assay demonstrated that bortezomib could induce apoptosis of LLPCs in a concentration-dependent manner. The proteasome activity assay showed that bortezomib significantly reduced the proteasome activity in sorted CD19CD138+ LLPCs. Furthermore, in active ITP murine models, bortezomib eliminated LLPCs in vivo and alleviated thrombocytopaenia. We conclude that LLPCs participate in the pathogenesis of ITP and bortezomib may have potential as a novel therapeutic regimen.

Authors' Contributions

G.L. and S.W. designed and performed research, analysed data and wrote the paper. N.L., Y.L., Q.F., X.L. and X.Z. performed research and analysed data. Y.H., C.M., L.S. and C.G. evaluated the data and corrected the article. M.H. and J.P. designed and performed research and reviewed the work. All authors read and edited the manuscript.


* Guosheng Li and Shuang Wang contributed equally to this study.


Supplementary Material

 
  • References

  • 1 Yang L, Wang L, Zhao CH. , et al. Contributions of TRAIL-mediated megakaryocyte apoptosis to impaired megakaryocyte and platelet production in immune thrombocytopenia. Blood 2010; 116 (20) 4307-4316
  • 2 Halliley JL, Tipton CM, Liesveld J. , et al. Long-lived plasma cells are contained within the CD19(-)CD38(hi)CD138(+) subset in human bone marrow. Immunity 2015; 43 (01) 132-145
  • 3 Jourdan M, Caraux A, Caron G. , et al. Characterization of a transitional preplasmablast population in the process of human B cell to plasma cell differentiation. J Immunol 2011; 187 (08) 3931-3941
  • 4 Gomez AM, Willcox N, Molenaar PC. , et al. Targeting plasma cells with proteasome inhibitors: possible roles in treating myasthenia gravis?. Ann N Y Acad Sci 2012; 1274: 48-59
  • 5 Radbruch A, Muehlinghaus G, Luger EO. , et al. Competence and competition: the challenge of becoming a long-lived plasma cell. Nat Rev Immunol 2006; 6 (10) 741-750
  • 6 Jourdan M, Cren M, Robert N. , et al. IL-6 supports the generation of human long-lived plasma cells in combination with either APRIL or stromal cell-soluble factors. Leukemia 2014; 28 (08) 1647-1656
  • 7 Ho F, Lortan JE, MacLennan IC, Khan M. Distinct short-lived and long-lived antibody-producing cell populations. Eur J Immunol 1986; 16 (10) 1297-1301
  • 8 Hiepe F, Dörner T, Hauser AE, Hoyer BF, Mei H, Radbruch A. Long-lived autoreactive plasma cells drive persistent autoimmune inflammation. Nat Rev Rheumatol 2011; 7 (03) 170-178
  • 9 Bortnick A, Chernova I, Quinn III WJ, Mugnier M, Cancro MP, Allman D. Long-lived bone marrow plasma cells are induced early in response to T cell-independent or T cell-dependent antigens. J Immunol 2012; 188 (11) 5389-5396
  • 10 Gomez AM, Willcox N, Vrolix K. , et al. Proteasome inhibition with bortezomib depletes plasma cells and specific autoantibody production in primary thymic cell cultures from early-onset myasthenia gravis patients. J Immunol 2014; 193 (03) 1055-1063
  • 11 Tran H, Brighton T, Grigg A. , et al. A multi-centre, single-arm, open-label study evaluating the safety and efficacy of fixed dose rituximab in patients with refractory, relapsed or chronic idiopathic thrombocytopenic purpura (R-ITP1000 study). Br J Haematol 2014; 167 (02) 243-251
  • 12 Kuter DJ, Macahilig C, Grotzinger KM. , et al. Treatment patterns and clinical outcomes in patients with chronic immune thrombocytopenia (ITP) switched to eltrombopag or romiplostim. Int J Hematol 2015; 101 (03) 255-263
  • 13 Neubert K, Meister S, Moser K. , et al. The proteasome inhibitor bortezomib depletes plasma cells and protects mice with lupus-like disease from nephritis. Nat Med 2008; 14 (07) 748-755
  • 14 Hoyer BF, Moser K, Hauser AE. , et al. Short-lived plasmablasts and long-lived plasma cells contribute to chronic humoral autoimmunity in NZB/W mice. J Exp Med 2004; 199 (11) 1577-1584
  • 15 Tiburzy B, Kulkarni U, Hauser AE, Abram M, Manz RA. Plasma cells in immunopathology: concepts and therapeutic strategies. Semin Immunopathol 2014; 36 (03) 277-288
  • 16 Tourigny MR, Ursini-Siegel J, Lee H. , et al. CDK inhibitor p18(INK4c) is required for the generation of functional plasma cells. Immunity 2002; 17 (02) 179-189
  • 17 Hauser AE, Debes GF, Arce S. , et al. Chemotactic responsiveness toward ligands for CXCR3 and CXCR4 is regulated on plasma blasts during the time course of a memory immune response. J Immunol 2002; 169 (03) 1277-1282
  • 18 Orlowski RZ, Kuhn DJ. Proteasome inhibitors in cancer therapy: lessons from the first decade. Clin Cancer Res 2008; 14 (06) 1649-1657
  • 19 Adams J. The proteasome: a suitable antineoplastic target. Nat Rev Cancer 2004; 4 (05) 349-360
  • 20 Hideshima T, Chauhan D, Richardson P. , et al. NF-kappa B as a therapeutic target in multiple myeloma. J Biol Chem 2002; 277 (19) 16639-16647
  • 21 Ichikawa HT, Conley T, Muchamuel T. , et al. Beneficial effect of novel proteasome inhibitors in murine lupus via dual inhibition of type I interferon and autoantibody-secreting cells. Arthritis Rheum 2012; 64 (02) 493-503
  • 22 Hainz N, Thomas S, Neubert K. , et al. The proteasome inhibitor bortezomib prevents lupus nephritis in the NZB/W F1 mouse model by preservation of glomerular and tubulointerstitial architecture. Nephron, Exp Nephrol 2012; 120 (02) e47-e58
  • 23 Nikolova-Ganeva KA, Gesheva VV, Todorov TA, Voll RE, Vassilev TL. Targeted silencing of DNA-specific B cells combined with partial plasma cell depletion displays additive effects on delaying disease onset in lupus-prone mice. Clin Exp Immunol 2013; 174 (02) 221-228
  • 24 Gomez AM, Vrolix K, Martínez-Martínez P. , et al. Proteasome inhibition with bortezomib depletes plasma cells and autoantibodies in experimental autoimmune myasthenia gravis. J Immunol 2011; 186 (04) 2503-2513
  • 25 van der Vlag J, Berden JH. Proteasome inhibition: a new therapeutic option in lupus nephritis?. Nephrol Dial Transplant 2008; 23 (12) 3771-3772
  • 26 Neunert C, Lim W, Crowther M, Cohen A, Solberg Jr L, Crowther MA. ; American Society of Hematology. The American Society of Hematology 2011 evidence-based practice guideline for immune thrombocytopenia. Blood 2011; 117 (16) 4190-4207
  • 27 Hou M, Peng J, Shi Y. , et al. Mycophenolate mofetil (MMF) for the treatment of steroid-resistant idiopathic thrombocytopenic purpura. Eur J Haematol 2003; 70 (06) 353-357
  • 28 Kiefel V, Santoso S, Weisheit M, Müeller-Eckhardt C. Monoclonal antibody--specific immobilization of platelet antigens (MAIPA): a new tool for the identification of platelet-reactive antibodies. Blood 1987; 70 (06) 1722-1726
  • 29 Balvan J, Krizova A, Gumulec J. , et al. Multimodal holographic microscopy: distinction between apoptosis and oncosis. PLoS One 2015; 10 (03) e0121674
  • 30 Wang R, Qin P, Ji XB, Hou M. The impact of platelet membrane autoantibodies on high-dose dexamethasone therapy in patients with idiopathic thrombocytopenic purpura [in Chinese]. Zhonghua Xue Ye Xue Za Zhi 2009; 30 (09) 619-621
  • 31 Auger S, Duny Y, Rossi JF, Quittet P. Rituximab before splenectomy in adults with primary idiopathic thrombocytopenic purpura: a meta-analysis. Br J Haematol 2012; 158 (03) 386-398
  • 32 Kim HJ, Krenn V, Steinhauser G, Berek C. Plasma cell development in synovial germinal centers in patients with rheumatoid and reactive arthritis. J Immunol 1999; 162 (05) 3053-3062
  • 33 Mahévas M, Michel M, Vingert B. , et al. Emergence of long-lived autoreactive plasma cells in the spleen of primary warm auto-immune hemolytic anemia patients treated with rituximab. J Autoimmun 2015; 62: 22-30
  • 34 Szyszko EA, Skarstein K, Jonsson R, Brokstad KA. Distinct phenotypes of plasma cells in spleen and bone marrow of autoimmune NOD.B10.H2b mice. Autoimmunity 2011; 44 (05) 415-426
  • 35 Taddeo A, Khodadadi L, Voigt C. , et al. Long-lived plasma cells are early and constantly generated in New Zealand Black/New Zealand White F1 mice and their therapeutic depletion requires a combined targeting of autoreactive plasma cells and their precursors. Arthritis Res Ther 2015; 17: 39
  • 36 Winter O, Dame C, Jundt F, Hiepe F. Pathogenic long-lived plasma cells and their survival niches in autoimmunity, malignancy, and allergy. J Immunol 2012; 189 (11) 5105-5111
  • 37 Chu VT, Berek C. The establishment of the plasma cell survival niche in the bone marrow. Immunol Rev 2013; 251 (01) 177-188
  • 38 Peperzak V, Vikström I, Walker J. , et al. Mcl-1 is essential for the survival of plasma cells. Nat Immunol 2013; 14 (03) 290-297
  • 39 McMillan R, Longmire RL, Yelenosky R, Lang JE, Heath V, Craddock CG. Immunoglobulin synthesis by human lymphoid tissues: normal bone marrow as a major site of IgG production. J Immunol 1972; 109 (06) 1386-1394
  • 40 Terstappen LW, Johnsen S, Segers-Nolten IM, Loken MR. Identification and characterization of plasma cells in normal human bone marrow by high-resolution flow cytometry. Blood 1990; 76 (09) 1739-1747
  • 41 Cogbill CH, Spears MD, Vantuinen P, Harrington AM, Olteanu H, Kroft SH. Morphologic and cytogenetic variables affect the flow cytometric recovery of plasma cell myeloma cells in bone marrow aspirates. Int J Lab Hematol 2015; 37 (06) 797-808
  • 42 Nadav L, Katz BZ, Baron S. , et al. Diverse niches within multiple myeloma bone marrow aspirates affect plasma cell enumeration. Br J Haematol 2006; 133 (05) 530-532
  • 43 Smock KJ, Perkins SL, Bahler DW. Quantitation of plasma cells in bone marrow aspirates by flow cytometric analysis compared with morphologic assessment. Arch Pathol Lab Med 2007; 131 (06) 951-955
  • 44 Kozutsumi Y, Segal M, Normington K, Gething MJ, Sambrook J. The presence of malfolded proteins in the endoplasmic reticulum signals the induction of glucose-regulated proteins. Nature 1988; 332 (6163): 462-464
  • 45 Kuhn DJ, Chen Q, Voorhees PM. , et al. Potent activity of carfilzomib, a novel, irreversible inhibitor of the ubiquitin-proteasome pathway, against preclinical models of multiple myeloma. Blood 2007; 110 (09) 3281-3290
  • 46 Csizmadia V, Raczynski A, Csizmadia E, Fedyk ER, Rottman J, Alden CL. Effect of an experimental proteasome inhibitor on the cytoskeleton, cytosolic protein turnover, and induction in the neuronal cells in vitro. Neurotoxicology 2008; 29 (02) 232-243
  • 47 Shin YK, Jang SY, Lee HK. , et al. Pathological adaptive responses of Schwann cells to endoplasmic reticulum stress in bortezomib-induced peripheral neuropathy. Glia 2010; 58 (16) 1961-1976
  • 48 Poruchynsky MS, Sackett DL, Robey RW, Ward Y, Annunziata C, Fojo T. Proteasome inhibitors increase tubulin polymerization and stabilization in tissue culture cells: a possible mechanism contributing to peripheral neuropathy and cellular toxicity following proteasome inhibition. Cell Cycle 2008; 7 (07) 940-949
  • 49 Bringhen S, Larocca A, Rossi D. , et al. Efficacy and safety of once-weekly bortezomib in multiple myeloma patients. Blood 2010; 116 (23) 4745-4753
  • 50 Moreau P, Pylypenko H, Grosicki S. , et al. Subcutaneous versus intravenous administration of bortezomib in patients with relapsed multiple myeloma: a randomised, phase 3, non-inferiority study. Lancet Oncol 2011; 12 (05) 431-440
  • 51 Tsukaguchi M, Shibano M, Matsuura A, Mukai S. The protective effects of lafutidine for bortezomib induced peripheral neuropathy. J Blood Med 2013; 4: 81-85