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Predictive value of galectin-1 in the development and progression of HIV-associated lymphoma

Vase, Maja Ølholma,*; Ludvigsen, Majaa,b,*; Bendix, Knudc; Dutoit, Stephen H.c; Hjortebjerg, Rikkeb; Petruskevicius, Irmaa; Møller, Michael B.d; Pedersen, Gittee; Denton, Paul W.b,f; Honoré, Bentg; Rabinovich, Gabriel A.h; Larsen, Carsten S.f; d’Amore, Francescoa

doi: 10.1097/QAD.0000000000001622
Correspondence
Free

aDepartment of Hematology, Aarhus University Hospital

bDepartment of Clinical Medicine, Aarhus University

cInstitute of Pathology, Aarhus University Hospital, Aarhus

dDepartment of Pathology, Odense University Hospital, Odense

eDepartment of Infectious Diseases, Aalborg University Hospital, Aalborg

fDepartment of Infectious Diseases, The Danish HIV Cohort, Aarhus University Hospital, Aarhus

gDepartment of Biomedicine, Aarhus University, Aarhus, Denmark

hLaboratorio de Inmunopatología, Instituto de Biología y Medicina Experimental (IBYME), Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina.

*Maja Ølholm Vase and Maja Ludvigsen equally contributed to this article.

Correspondence to Maja Ølholm Vase, MD PhD, Department of Hematology, Aarhus University Hospital, Tage Hansensgade 2, DK-8000 Aarhus C, Denmark. Tel: +45 2721 9330; fax: +45 467598; e-mail: majvas@rm.dk

Received 14 May, 2017

Revised 24 July, 2017

Accepted 8 August, 2017

At HIV-1 infection, the binding of the viral envelope proteins to CD4+ is essential for viral transmission, and this process is facilitated by interaction with the highly conserved host lectin, galectin-1 (Gal-1) [1–3]. Within the tumor microenvironment, Gal-1 is expressed by both tumor and stromal cells where it promotes tumor immune escape and favors hypoxia-driven angiogenesis [4–6]. In sporadically occurring Hodgkin lymphoma, high Gal-1 expression at diagnosis is associated with poorer treatment response [7], and high soluble Gal-1 (sGal-1) correlates with adverse disease characteristics [8]. Previous studies have shown that targeted inhibition of Gal-1 prevents tumor-induced immunosuppression [9,10] and inhibits tumor growth and metastasis in various tumor models [6,11–13].

Recently, we published a proteomic profiling study of pretreatment serum samples from HIV-infected patients, identifying several differentially expressed proteins associated with lymphoma development [14]. In this cohort, we have now evaluated serum levels of sGal-1 and correlated this with clinical parameters, including lymphoma development. In addition, we have investigated the intratumoral expression and prognostic value of Gal-1 in HIV-associated lymphomas, and, for comparison, sGal-1 serum levels in 30 healthy blood donors [15]

Circulating sGal-1 levels were measured using a time-resolved immunofluorometric assay and immunohistochemistry and the evaluation of tumoral Gal-1 expression were performed as described previously [7,14,15].

Pretreatment sGal-1 serum levels were assessed in 19 HIV-positive individuals at time of HIV diagnosis. There were no sex-related differences (P = 0.450) and sGal-1 levels neither correlate with peripheral CD4+ cell count nor with viral load at HIV diagnosis (ρ = −0.491 P = 0.852 and ρ = −0.009 P = 0.974, respectively).

HIV-infected individuals had significantly lower levels of sGal-1 compared with healthy controls (43.6 vs. 84.9 ng/ml; P < 0.001; Fig. 1a). Within the entire study cohort (healthy controls and HIV-infected individuals), those patients who would later develop lymphoma also had significantly lower levels of sGal-1 at time of HIV-diagnosis (Fig. 1b; P = 0.016). There was no significant difference in sGal-1 within the HIV cohort (Fig. 1c, P = 0.130).

Fig. 1

Fig. 1

A cut-off value of 2.4 ng/ml generated by receiver operating curve (ROC) analysis separated HIV-infected individuals who later developed lymphoma from the remaining cohort of HIV patients and controls with a specificity of 82% and a sensitivity of 100%. Based on this cut-off value, 13 (31%) HIV-infected patients were allocated to the low sGal-1 group, including all future lymphoma patients (N = 5).

Tumoral Gal-1 expression correlated positively with a proinflammatory signature of the microenvironment, including the macrophage marker CD68, the cytotoxic markers CD8 and granzyme B, as well as the activation marker CD30 [CD68 (ρ = 0.740; P < 0.001), CD8 (ρ = 0.379; P = 0.027), granzyme B (ρ = 0.579; P < 0.001) and CD30 (ρ = 0.467; P = 0.006)]. Clinical features of the cohort included in the tissue microarray have been described previously [14].

Gal-1 was widely expressed in all lymphoma subtypes. Based on a ROC generated cut-off value for high vs. low intratumoral Gal-1 expression, 59% (N = 10) of diffuse large B-cell lymphoma (DLBCL) patients had a high level of intratumoral Gal-1 expression (>24.8% positive cells). In the total lymphoma cohort (all diagnoses), two-thirds of the patients (N = 22; 65%) were high expressers. This latter group more often had nodal disease and B-symptoms (P = 0.006). Gal-1 did not correlate with tumoral Epstein–Barr virus (EBV) status, EBV latency type, international prognostic index (IPI), clinical stage, or cell of origin.

In HIV-associated DLBCL, patients with higher levels of intratumoral Gal-1 expression had a significantly better outcome with a 5-year overall survival of 70.0% (95% confidence interval 32.9–89.2%) vs. 14.3% (95% confidence interval 0.7–46.5%). In a multivariate analysis, adjusting for IPI and rituximab treatment, both Gal-1 expression (P = 0.021) and IPI (P = 0.049) retained an independent prognostic value.

HIV infection has a profound influence on the host immune system including altered cytokine and protein expression years prior to lymphoma diagnosis [14,16–18]. Gal-1 is secreted by most immune cells [19] and the significantly lower levels of sGal-1 in newly diagnosed HIV-infected individuals (compared with healthy controls), as found in our study, may reflect the dramatically altered immune constitution of these patients. This may lead to a proinflammatory although nonefficient T-cell response, ultimately leading to lymphoma development.

We found a relatively high intratumoral expression of Gal-1 in our cohort of HIV-associated DLBCL, as compared with the immunocompetent setting [20]. This may partly reflect different evaluation techniques, but inherent disparities in lymphoma microenvironment may also be involved [21,22]. Gal-1 is largely produced by macrophages [23]. The correlation between high Gal-1 expression and improved outcome in HIV-associated DLBCL may therefore be explained by a higher level of macrophages because they have been shown to improve the efficacy of antibody-driven immunotherapy [24].

In conclusion, the results of our study indicate that Gal-1 is significantly associated with risk of lymphoma in HIV-infected individuals and may represent an attractive future target for the management of HIV-associated lymphoma.

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Acknowledgements

The authors wish to thank Erik Hagen Nielsen, Vibeke Ellerup Jensen, and Kristina Lystlund Lauridsen for expert technical assistance, and Betina S. Sørensen for facilitating access to healthy donors samples.

Conceived and designed the study: M.Ø.V., M.L., C.S.L., F.d’A. Provided study material: G.A.R., I.P., G.P., C.S.L., M.B.M., K.B., S.H.D. Performed the experiments: R.H., M.L. Analyzed data: M.Ø.V., M.L., B.H., P.W.D. Wrote the paper: M.Ø.V. and M.L. Final editing and approval of the manuscript: all authors.

The work was supported by unrestricted grants from Dagmar Marshalls Foundation, Manufacturer Einer Willumsen's Memorial Foundation, The Harboe Foundation, The Krista and Viggo Petersens Foundation, Fonden til Lægevidenskabens fremme, Director Emil C. Hertz and his wife Inger Hertz Foundation, The Foundation of 17 December 1981, Architect Holger Hjortenberg and wife Dagmar Hjortenberg's Foundation; Frits, Georg, and Marie Cecilie Glud's Foundation, Danish Diabetes Academy supported by the Novo Nordisk Foundation, Inger and Max Wørzner's Memorial Foundation, and The MEMBRANES Center at Aarhus University.

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Conflicts of interest

There are no conflicts of interest.

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References

1. Ouellet M, Mercier S, Pelletier I, Bounou S, Roy J, Hirabayashi J, et al. Galectin-1 acts as a soluble host factor that promotes HIV-1 infectivity through stabilization of virus attachment to host cells. J Immunol 2005; 174:4120–4126.
2. Mercier S, St-Pierre C, Pelletier I, Ouellet M, Tremblay MJ, Sato S. Galectin-1 promotes HIV-1 infectivity in macrophages through stabilization of viral adsorption. Virology 2008; 371:121–129.
3. St-Pierre C, Manya H, Ouellet M, Clark GF, Endo T, Tremblay MJ, Sato S. Host-soluble galectin-1 promotes HIV-1 replication through a direct interaction with glycans of viral gp120 and host CD4. J Virol 2011; 85:11742–11751.
4. Juszczynski P, Ouyang J, Monti S, Rodig SJ, Takeyama K, Abramson J, et al. The AP1-dependent secretion of galectin-1 by Reed Sternberg cells fosters immune privilege in classical Hodgkin lymphoma. Proc Natl Acad Sci U S A 2007; 104:13134–13139.
5. Cedeno-Laurent F, Watanabe R, Teague JE, Kupper TS, Clark RA, Dimitroff CJ. Galectin-1 inhibits the viability, proliferation, and Th1 cytokine production of nonmalignant T cells in patients with leukemic cutaneous T-cell lymphoma. Blood 2012; 119:3534–3538.
6. Croci DO, Salatino M, Rubinstein N, Cerliani JP, Cavallin LE, Leung HJ, et al. Disrupting galectin-1 interactions with N-glycans suppresses hypoxia-driven angiogenesis and tumorigenesis in Kaposi's sarcoma. J Exp Med 2012; 209:1985–2000.
7. Kamper P, Ludvigsen M, Bendix K, Hamilton-Dutoit S, Rabinovich GA, Moller MB, et al. Proteomic analysis identifies galectin-1 as a predictive biomarker for relapsed/refractory disease in classical Hodgkin lymphoma. Blood 2011; 117:6638–6649.
8. Ouyang J, Plütschow A, Pogge von Strandmann E, Reiners KS, Ponader S, Rabinovich GA, et al. Galectin-1 serum levels reflect tumor burden and adverse clinical features in classical Hodgkin lymphoma. Blood 2013; 121:3431–3433.
9. Rubinstein N, Alvarez M, Zwirner NW, Toscano MA, Ilarregui JM, Bravo A, et al. Targeted inhibition of galectin-1 gene expression in tumor cells results in heightened T cell-mediated rejection; a potential mechanism of tumor-immune privilege. Cancer Cell 2004; 5:241–251.
10. Ouyang J, Juszczynski P, Rodig SJ, Green MR, O’Donnell E, Currie T, et al. Viral induction and targeted inhibition of galectin-1 in EBV+ posttransplant lymphoproliferative disorders. Blood 2011; 117:4315–4322.
11. Dalotto-Moreno T, Croci DO, Cerliani JP, Martinez-Allo VC, Dergan-Dylon S, Méndez-Huergo SP, et al. Targeting galectin-1 overcomes breast cancer-associated immunosuppression and prevents metastatic disease. Cancer Res 2013; 73:1107–1117.
12. Laderach DJ, Gentilini LD, Giribaldi L, Delgado VC, Nugnes L, Croci DO, et al. A unique galectin signature in human prostate cancer progression suggests galectin-1 as a key target for treatment of advanced disease. Cancer Res 2013; 73:86–96.
13. Croci DO, Cerliani JP, Dalotto-Moreno T, Méndez-Huergo SP, Mascanfroni ID, Dergan-Dylon S, et al. Glycosylation-dependent lectin-receptor interactions preserve angiogenesis in anti-VEGF refractory tumors. Cell 2014; 156:744–758.
14. Vase MØ, Ludvigsen M, Bendix K, Hamilton-Dutoit S, Mller MB, Pedersen C, et al. Proteomic profiling of pretreatment serum from HIV-infected patients identifies candidate markers predictive of lymphoma development. AIDS 2016; 30:1889–1898.
15. Petruskevicius I, Ludvigsen M, Hjortebjerg R, Sørensen BS, Kamper P, Vase M, et al. Clinical relevance of galectin-1 in hematologic malignancies treated with non-myeloablative hemopoietic stem cell transplantation. Bone Marrow Transplant 2016; 51:1387–1390.
16. Rabkin CS, Engels EA, Landgren O, Schuurman R, Camargo MC, Pfeiffer R, Goedert JJ. Circulating cytokine levels, Epstein-Barr viremia and risk of acquired immunodeficiency syndrome-related non-Hodgkin lymphoma. Am J Hematol 2011; 86:875–878.
17. Breen EC, Hussain SK, Magpantay L, Jacobson LP, Detels R, Rabkin CS, et al. B-cell stimulatory cytokines and markers of immune activation are elevated several years prior to the diagnosis of systemic AIDS-associated non-Hodgkin B-cell lymphoma. Cancer Epidemiol Biomarkers Prev 2011; 20:1303–1314.
18. Hussain SK, Hessol NA, Levine AM, Breen EC, Anastos K, Cohen M, et al. Serum biomarkers of immune activation and subsequent risk of non-Hodgkin B-cell lymphoma among HIV-infected women. Cancer Epidemiol Biomark Prev 2013; 22:2084–2093.
19. Toscano MA, Campagna L, Molinero LL, Cerliani JP, Croci DO, Ilarregui JM, et al. Nuclear factor (NF)-kB controls expression of the immunoregulatory glycan-binding protein galectin-1. Mol Immunol 2011; 48:1940–1949.
20. Rodig SJ, Ouyang J, Juszczynski P, Currie T, Law K, Neuberg DS, et al. AP1-dependent galectin-1 expression delineates classical Hodgkin and anaplastic large cell lymphomas from other lymphoid malignancies with shared molecular features. Clin Cancer Res 2008; 14:3338–3344.
21. Liapis K, Clear A, Owen A, Coutinho R, Greaves P, Lee AM, et al. The microenvironment of AIDS-related diffuse large B-cell lymphoma provides insight into the pathophysiology and indicates possible therapeutic strategies. Blood 2013; 122:424–433.
22. Chao C, Silverberg MJ, Xu L, Chen LH, Castor B, Martínez-Maza O, et al. A comparative study of molecular characteristics of diffuse large B-cell lymphoma from patients with and without human immunodeficiency virus infection. Clin Cancer Res 2015; 21:1429–1437.
23. Barrionuevo P, Beigier-Bompadre M, Ilarregui JM, Toscano MA, Bianco GA, Isturiz MA, Rabinovich GA. A novel function for galectin-1 at the crossroad of innate and adaptive immunity: galectin-1 regulates monocyte/macrophage physiology through a nonapoptotic ERK-dependent pathway. J Immunol 2007; 178:436–445.
24. Riihijärvi S, Fiskvik I, Taskinen M, Vajavaara H, Tikkala M, Yri O, et al. Prognostic influence of macrophages in patients with diffuse large B-cell lymphoma: a correlative study from a Nordic phase II trial. Haematologica 2015; 100:238–245.
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