Targeting the polarization of tumor-associated macrophages and modulating mir-155 expression might be a new approach to treat diffuse large B-cell lymphoma of the elderly
Aging immune deterioration and Epstein–Barr (EBV) intrinsic mechanisms play an essential role in EBV-positive diffuse large B-cell lymphoma (DLBCL) of the elderly (EBV + DLBCLe) pathogenesis, through the expression of viral proteins, interaction with host molecules and epigenetic regulation, such as miR-155, required for induction of M1 phenotype of macrophages. This study aims to evaluate the relationship between macrophage polarization pattern in the tumor microenvironment and relative expression of miR-155 in EBV + DLBCLe and EBV-negative DLBCL patients. We studied 28 EBV + DLBCLe and 65 EBV-negative DLBCL patients. Tumor-associated macrophages (TAM) were evaluated by expression of CD68, CD163 and CD163/CD68 ratio (degree of M2 polarization), using tissue microarray. RNA was extracted from paraffin-embedded tumor samples for miR-155 relative expression study. We found a significantly higher CD163/CD68 ratio in EBV + DLBCLe compared to EBV-negative DLBCL. In EBV-negative DLBCL, CD163/CD68 ratio was higher among advanced-staged/high-tumor burden disease and overexpression of miR-155 was associated with decreased polarization to the M2 phenotype of macrophages. The opposite was observed in EBV + DLBCLe patients: we found a positive association between miR-155 relative expression and CD163/CD68 ratio, which was not significant after outlier exclusion. We believe that the higher CD163/CD68 ratio in this group is probably due to the presence of the EBV since it directly affects macrophage polarization towards M2 phenotype through cytokine secretion in the tumor microenvironment. Therapeutic strategies modulating miR-155 expression or preventing immuno-regulatory and pro-tumor macrophage polarization could be adjuvants in EBV + DLBCLe therapy since this entity has a rich infiltration of M2 macrophages in its tumor microenvironment.
KeywordsNon-Hodgkin lymphoma EBV Immunohistochemistry Tumor-associated macrophages microRNA
Diffuse large B-cell lymphoma
Diffuse large B-cell lymphoma of the elderly
Formalin-fixed paraffin embedded
International Prognostic Index
Quantitative real-time polymerase chain reaction
Rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone
Wagner A. Poles and Gisele W. B. Colleoni designed the work, acquisition, analysis, and interpretation of data, wrote the manuscript; Erika E. Nishi, Tathiana A. de Andrade, and Ruy R. de Campos Jr. analyzed data and critically reviewed the manuscript for important intellectual content; Mariana B. de Oliveira and Angela I. P. Eugênio wrote and critically reviewed the manuscript; Antonio Hugo F. M. Campos, Gilles Landman, José Vassallo, Cristovam Scapulatempo Neto, Roberto Antonio Pinto Paes, and Antonio C. Alves reviewed samples, analyzed data and critically reviewed the manuscript; Maria Cláudia N. Zerbini contributed substantially to the conception, analyzed data, and critically reviewed the work.
Wagner A. Poles was partially supported by Fundaçao de Amparo à Pesquisa do Estado de Sao Paulo (FAPESP), Brazil. This work was supported by Fundaçao de Amparo à Pesquisa do Estado de Sao Paulo (FAPESP), Brazil 2010/17668-6.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest to disclose.
This study was approved by the ethical committees of all collaborating centers: Department of Clinical and Experimental Oncology, Universidade Federal de São Paulo, Brazil; Department of Physiology, Universidade Federal de São Paulo, Brazil; Department of Pathology, AC Camargo Cancer Center, Brazil; Department of Pathology, Universidade Federal de São Paulo, Brazil; Molecular Oncology Research Center, Barretos Cancer Hospital, Brazil; Faculty of Medical Sciences of Santa Casa of São Paulo; Department of Pathology, Universidade de São Paulo, Brazil, and was in accordance with the ethical standards of the institutional committees and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
Informed consent was obtained from all individual participants included in the study.
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