Abstract
DLBCL is the most common lymphoma subtype occurring in older populations as well as in younger HIV infected patients. The current treatment options for DLBCL are effective for most patients yet the relapse rate is high. While many biomarkers for DLBCL exist, they are not in clinical use due to low sensitivity and specificity. In addition, these biomarkers have not been studied in the HIV context. Therefore, the identification of new biomarkers for HIV negative and HIV positive DLBCL, may lead to a better understanding of the disease pathology and better therapeutic design. Protein biomarkers for DLBCL were determined using MALDI imaging mass spectrometry (IMS) and characterised using LC–MS. The expression of one of the biomarkers, heat shock protein (Hsp) 70, was confirmed on a separate cohort of samples using immunohistochemistry. The biomarkers identified in the study consisted of four protein clusters including glycolytic enzymes, ribosomal proteins, histones and collagen. These proteins could differentiate between control and tumour tissue, and the DLBCL immunohistochemical subtypes in both cohorts. The majority (41/52) of samples in the confirmation cohort were negative for Hsp70 expression. The HIV positive DLBCL cases had a higher percentage of cases expressing Hsp70 than their HIV negative counterparts. The non-GC subtype also frequently overexpressed Hsp70, confirming MALDI IMS data. The expression of Hsp70 did not correlate with survival in both the HIV negative and HIV positive cohort. This study identified potential biomarkers for HIV negative and HIV positive DLBCL from FFPE tissue sections. These may be used as diagnostic and prognostic markers complementary to current clinical management programmes for DLBCL.
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References
Álvarez-chaver P, Otero-estévez O, Páez M et al (2014) Proteomics for discovery of candidate colorectal cancer biomarkers. World J Gastroenterol 20:3804–3824. doi:10.3748/wjg.v20.i14.3804
Baijnath S, Naiker S, Shobo A et al (2015) Evidence for the presence of clofazimine and its distribution in the healthy mouse brain. J Mol Histol 46:439–442. doi:10.1007/s10735-015-9634-3
Balluff B, Schöne C, Höfler H, Walch A (2011) MALDI imaging mass spectrometry for direct tissue analysis: technological advancements and recent applications. Histochem Cell Biol 136:227–244. doi:10.1007/s00418-011-0843-x
Balluff B, Frese CK, Maier SK et al (2015) De novo discovery of phenotypic intratumour heterogeneity using imaging mass spectrometry. J Pathol 235:3–13. doi:10.1002/path.4436
Baptista MJ, Garcia O, Morgades M et al (2015) HIV-infection impact on clinical–biological features and outcome of diffuse large B-cell lymphoma treated with R-CHOP in the combination antiretroviral therapy era. AIDS 29:811–818. doi:10.1097/QAD.0000000000000624
Barbini L, Rodrı J, Dominguez F, Vega F (2007) Glyceraldehyde-3-phosphate dehydrogenase exerts different biologic activities in apoptotic and proliferating hepatocytes according to its subcellular localization. Mol Cell Biochem 300:19–28. doi:10.1007/s11010-006-9341-1
Barnard G, Mori M, Staniunas R et al (1995) Ubiquitin fusion proteins are overexpressed in colon cancer but not in gastric cancer. Biochim Biophys Acta 1272:147–153
Basso K, Dalla-favera R (2015) Germinal centres and B cell lymphomagenesis E l. Nat Publ Gr 15:172–184. doi:10.1038/nri3814
Caprioli RM, Farmer TB, Gile J (1997) Molecular imaging of biological samples: localization of peptides and proteins using MALDI-TOF MS. Anal Chem 69:4751–4760
Carbone A, Gloghini A (2014) Diffuse large B cell lymphoma: using pathologic and molecular biomarkers to define subgroups for novel therapy. Ann Hematol 93:1263–1277. doi:10.1007/s00277-014-2116-y
Castellino S, Groseclose MR, Wagner D (2011) MALDI imaging mass spectrometry: bridging biology and chemistry in drug development. Bioanalysis 3:2427–2441. doi:10.4155/bio.11.232
Chadburn A, Chiu A, Lee JY et al (2009) Immunophenotypic analysis of AIDS-related diffuse large B-cell lymphoma and clinical implications in patients from AIDS Malignancies Consortium clinical trials 010 and 034. J Clin Oncol 27:5039–5048. doi:10.1200/JCO.2008.20.5450
Chiche J, Pommier S, Beneteau M et al (2015) GAPDH enhances the aggressiveness and the vascularization of non-Hodgkin’s B lymphomas via NF-κB-dependent induction of HIF-1α. Leukemia 29:1163–1176. doi:10.1038/leu.2014.324
Coutinho R, Pria AD, Gandhi S et al (2014) HIV status does not impair the outcome of patients diagnosed with diffuse large B-cell lymphoma treated with R-CHOP in the cART era. AIDS 28:689–697. doi:10.1097/QAD.0000000000000133
Davis RE, Brown KD, Siebenlist U, Staudt LM (2001) Constitutive nuclear factor KB activity is required for survival of activated B cell—like diffuse large B cell lymphoma cells. J Exp Med 194:1861–1874
Deeb SJ, D’Souza RCJ, Cox J et al (2012) Super-SILAC allows classification of diffuse large B-cell lymphoma subtypes by their protein expression profiles. Mol Cell Proteomics 11:77–89. doi:10.1074/mcp.M111.015362
Diaz-Ramos A, Roig-borrellas A, Garc A, Roser L (2012) α -Enolase, a Multifunctional Protein: Its Role on Pathophysiological Situations. J Biomed Biotechnol p 12. doi: 10.1155/2012/156795
Evans LS, Hancock BW (2003) Non-Hodgkin lymphoma. Lancet 362:139–146. doi:10.1016/S0140-6736(03)13868-8
Fang X, Jiang Y, Feng L et al (2013) Blockade of PI3 K/AKT pathway enhances sensitivity of Raji cells to chemotherapy through down-regulation of HSP70. Cancer Cell Int 13:1. doi:10.1186/1475-2867-13-48
Gloghini A, Dolcetti R, Carbone A (2013) Lymphomas occurring specifically in HIV-infected patients: from pathogenesis to pathology. Semin Cancer Biol 23:457–467. doi:10.1016/j.semcancer.2013.08.004
Han X, Lee M, Yu G et al (2012) Altered dynamics of ubiquitin hybrid proteins during tumor cell apoptosis. Cell Death Dis 3:e255. doi:10.1038/cddis.2011.142
Hans CP, Weisenburger DD, Greiner TC et al (2004) Confirmation of the molecular classification of diffuse large B-cell lymphoma by immunohistochemistry using a tissue microarray. Blood 103:275–282. doi:10.1182/blood-2003-05-1545
Henry JL, Coggin DL, King CR (1993) High-level expression of the ribosomal protein L19 in human breast tumors that overexpress erbB-21. Cancer Res 53:1403–1408
Hunt KE, Reichard KK (2008) Diffuse large B-cell lymphoma. Arch Pathol Lab Med 132:118–124. doi:10.1043/1543-2165(2008)132[118:DLBL]2.0.CO;2
Hwang S-I, Thumar J, Lundgren DH et al (2007) Direct cancer tissue proteomics: a method to identify candidate cancer biomarkers from formalin-fixed paraffin-embedded archival tissues. Oncogene 26:65–76. doi:10.1038/sj.onc.1209755
Israelsen WJ, Vander MG (2015) Seminars in Cell & Developmental Biology Pyruvate kinase: function, regulation and role in cancer. Semin Cell Dev Biol 43:43–51. doi:10.1016/j.semcdb.2015.08.004
Jiang B, Liang P, Deng G et al (2011) Increased stability of Bcl-2 in HSP70-mediated protection against apoptosis induced by oxidative stress. Cell Stress Chaperones 16:143–152. doi:10.1007/s12192-010-0226-6
Kasai H, Nadano D, Hidaka E, Higuchi K, Kawakubo M, Sato T-A, Nakayama J (2003) Differential expression of ribosomal proteins in human normal and neoplastic colorectum. J Histochem Cytochem 51(5):567–573
Kreunin P, Yoo C, Urquidi V et al (2007) Differential expression of ribosomal proteins in a human metastasis model identified by coupling 2-D liquid chromatography and mass spectrometry. Cancer Genomics Proteomics 4:329–340
Kumari S, Malla R (2015) New insight on the role of plasminogen receptor in cancer progression. Cancer Growth Metastasis 8:35–42. doi:10.4137/CGM.S27335.TYPE
Lee HW, Lee EH, Kim S-H et al (2013) Heat shock protein 70 (HSP70) expression is associated with poor prognosis in intestinal type gastric cancer. Virchows Arch 463:489–495. doi:10.1007/s00428-013-1461-x
Leopardi O, Naughten W, Giannulis I et al (2001) HSP70 is selectively over-expressed in the blast cells of the germinal centres and paracortex in reactive lymph nodes. Histopathology 39:566–571
Liapis K, Clear A, Owen A et al (2013) The microenvironment of AIDS-related diffuse large B-cell lymphoma provides insight into the pathophysiology and indicates possible therapeutic strategies. Blood 122:424–433. doi:10.1182/blood-2013-03-488171
Liu W, Chen Y, Lu G et al (2011) Down-regulation of HSP70 sensitizes gastric epithelial cells to apoptosis and growth retardation triggered by H. pylori. BMC Gastroenterol 11:146. doi:10.1186/1471-230X-11-146
Liu Y, Zeng L, Zhang S et al (2013) Identification of differentially expressed proteins in chemotherapy-sensitive and chemotherapy-resistant diffuse large B cell lymphoma by proteomic methods. Med Oncol 30:528. doi:10.1007/s12032-013-0528-5
López-Guillermo A, Colomo L, Jiménez M et al (2005) Diffuse large B-cell lymphoma: clinical and biological characterization and outcome according to the nodal or extranodal primary origin. J Clin Oncol 23:2797–2804. doi:10.1200/JCO.2005.07.155
Mafune K, Wong JM, Staniunas RJ et al (1991) Ubiquitin hybrid protein gene expression during human colon cancer progression. Arch Surg 126:462–466
Mao-de L, Jing X (2007) Ribosomal proteins and colorectal cancer. Curr Genomics 8:43–49
Marín-Hernández A, Gallardo-Pérez JC, Rodríguez-Enríquez S et al (2011) Modeling cancer glycolysis. Biochim Biophys Acta 1807:755–767. doi:10.1016/j.bbabio.2010.11.006
Marino-Ramirez L, Kann MG, Shoemaker BA, Landsman D (2005) Histone structure and nucleosome stability. Expert Rev Proteomics 2:719–729
Martinez-balibrea E, Plasencia C, Ginés A et al (2009) A proteomic approach links decreased pyruvate kinase M2 expression to oxaliplatin resistance in patients with colorectal cancer and in human cell lines. Mol Cancer Ther 8:771–779. doi:10.1158/1535-7163.MCT-08-0882
Mey U, Hitz F, Lohri A et al (2012) Diagnosis and treatment of diffuse large B-cell lymphoma. Swiss Med Wkly 142:w13511. doi:10.4414/smw.2012.13511
Mjahed H, Girodon F, Fontenay M, Garrido C (2012) Heat shock proteins in hematopoietic malignancies. Exp Cell Res 318:1946–1958. doi:10.1016/j.yexcr.2012.05.012
Montes-Moreno S, Gonzalez-Medina A-R, Rodriguez-Pinilla S-M et al (2010) Aggressive large B-cell lymphoma with plasma cell differentiation: immunohistochemical characterization of plasmablastic lymphoma and diffuse large B-cell lymphoma with partial plasmablastic phenotype. Haematologica 95:1342–1349. doi:10.3324/haematol.2009.016113
Muller P, Ruckova E, Halada P et al (2013) C-terminal phosphorylation of Hsp70 and Hsp90 regulates alternate binding to co-chaperones CHIP and HOP to determine cellular protein folding/degradation balances. Oncogene 32:3101–3110
Murphy ME (2013) The HSP70 family and cancer. Carcinogenesis 34:1181–1188. doi:10.1093/carcin/bgt111
Nanbu K, Konishi M, Mandai M et al (1998) Prognostic significance of heat shock proteins HSP70 and HSP90 in endometrial carcinomas. Cancer Detect Prev 22:549–555
Pancholi V (2001) Multifunctional alpha-enolase: its role in diseases. Cell Mol Life Sci 58:902–920
Pontén F, Jirström K, Uhlen M (2008) The human protein Atlas—a tool for pathology. J Pathol 216:387–393. doi:10.1002/path.2440
Qi H, Yukun ZU, Xiangning FU, Tangchun WU (2005) Expression of heat shock protein 70 and 27 in non-small cell lung cancer and its clinical significance. J Huazhong Univ Sci Technol 25:693–695
Rauser S, Marquardt C, Balluff B et al (2010) Classification of HER2 receptor status in breast cancer tissues by MALDI imaging mass spectrometry. J Proteome Res 9:1854–1863. doi:10.1021/pr901008d
Reyzer ML, Caprioli RM (2005) MALDI mass spectrometry for direct tissue analysis: a new tool for biomarker discovery. J Proteome Res 4:1138–1142. doi:10.1021/pr050095+
Rickert RC (2013) New insights into pre-BCR and BCR signalling with relevance to B cell malignancies. Nat Rev Immunol 13:578–591. doi:10.1038/nri3487
Rohde M, Daugaard M, Jensen MH et al (2005) Members of the heat-shock protein 70 family promote cancer cell growth by distinct mechanisms. Genes Dev 19:570–582. doi:10.1101/gad.305405
Rosenwald A, Wright G, Chan WC et al (2002) The use of molecular profiling to predict survival after chemotherapy for diffuse large-B-cell lymphoma. New Engl J Med 346:1937–1947. doi:10.1056/NEJMoa012914
Rüetschi U, Stenson M, Hasselblom S, et al (2015) SILAC-based quantitative proteomic analysis of diffuse large B-cell lymphoma patients. Int J Proteomics 2015: p 12
Schöne C, Höfler H, Walch A (2013) MALDI imaging mass spectrometry in cancer research: combining proteomic profiling and histological evaluation. Clin Biochem 46:539–545. doi:10.1016/j.clinbiochem.2013.01.018
Shaffer AL, Rosenwald A, Staudt LM (2002) Lymphoid malignancies: the dark side of B-cell differentiation. Nat Rev Immunol 2:920–932. doi:10.1038/nri953
Staudt LM, Dave S (2005) The biology of human lymphoid malignancies revealed by gene expression profiling. Adv Immunol 87:163–208. doi:10.1016/S0065-2776(05)87005-1
Tanaka M, Mun S, Harada A et al (2014) Hsc70 contributes to cancer cell survival by preventing Rab1A degradation under stress conditions. PLoS One 9:e96785. doi:10.1371/journal.pone.0096785
Tang Z, Yuan S, Hu Y et al (2012) Over-expression of GAPDH in human colorectal carcinoma as a prefered target of 3-Bromopyruvate propyl ester. J Bioenergy Biomembr 44:117–125. doi:10.1007/s10863-012-9420-9
Uhlén M, Björling E, Agaton C et al (2005) A human protein atlas for normal and cancer tissues based on antibody proteomics. Mol Cell Proteom 4:1920–1932. doi:10.1074/mcp.M500279-MCP200
Wang H, Zhao L, Li K et al (2006) Overexpression of ribosomal protein L15 is associated with cell proliferation in gastric cancer. BMC Cancer 6:91. doi:10.1186/1471-2407-6-91
Yoo BC, Ku J-L, Hong S-H et al (2004) Dcreased Pyruvate kinase M2 activity linked to Cisplatin resistance in human gastric carcinoma cell lines. Int J Cancer 108:532–539. doi:10.1002/ijc.11604
Young JC, Barral M, Hartl FU (2003) More than folding: localized functions of cytosolic chaperones. Trends Biochem Sci 28:541–547. doi:10.1016/j.tibs.2003.08.009
Zhang X, Wang B, Zhang X et al (2007) Serum diagnosis of diffuse large B-cell lymphomas and further identification of response to therapy using SELDI-TOF-MS and tree analysis patterning. BMC Cancer 7:235. doi:10.1186/1471-2407-7-235
Zhou D, Xie W, Hu K et al (2013) Prognostic values of various clinical factors and genetic subtypes for diffuse large B-cell lymphoma patients: a retrospective analysis of 227 cases. Asian Pac J Cancer Prev 14:929–934
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The work was supported by grants from the National Research Foundation, Cancer Association of South Africa, National Health Laboratory Service Research Trust and the University of Cape Town.
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Magangane, P., Sookhayi, R., Govender, D. et al. Determining protein biomarkers for DLBCL using FFPE tissues from HIV negative and HIV positive patients. J Mol Hist 47, 565–577 (2016). https://doi.org/10.1007/s10735-016-9695-y
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DOI: https://doi.org/10.1007/s10735-016-9695-y