[en] The lymph node (LN) pre-metastatic niche is faintly characterized in lymphophilic human neoplasia, although LN metastasis is considered as the strongest prognostic marker of patient survival. Due to its specific dissemination through a complex bilateral pelvic lymphatic system, early cervical cancer is a relevant candidate for investigating the early nodal metastatic process. In the present study, we analyzed in-depth both the lymphatic vasculature and the immune climate of pre-metastatic sentinel LN (SLN), in 48 cases of FIGO stage IB1 cervical neoplasms. An original digital image analysis methodology was used to objectively determine whole slide densities and spatial distributions of immunostained structures. We observed a marked increase in lymphatic vessel density (LVD) and a specific capsular and subcapsular distribution in pre-metastatic SLN when compared with non-sentinel counterparts. Such features persisted in the presence of nodal metastatic colonization. The inflammatory profile attested by CD8+, Foxp3, CD20 and PD-1expression was also significantly increased in pre-metastatic SLN. Remarkably, the densities of CD20+ B cells and PD-1 expressing germinal centers were positively correlated with LVD. All together, these data strongly support the existence of a pre-metastatic dialog between the primary tumor and the first nodal relay. Both lymphatic and immune responses contribute to the elaboration of a specific pre-metastatic microenvironment in human SLN. Moreover, this work provides evidence that, in the context of early cervical cancer, a pre-metastatic lymphangiogenesis occurs within the SLN (pre-metastatic niche) and is associated with a specific humoral immune response.
Disciplines :
Oncology
Author, co-author :
Balsat, Cédric ; Centre Hospitalier Universitaire de Liège - CHU > Centre d'oncologie
Blacher, Silvia ; Université de Liège > Département des sciences cliniques > Labo de biologie des tumeurs et du développement
Herfs, Michael ; Université de Liège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
Van De Velde, Maureen ; Université de Liège > Département des sciences cliniques > Labo de biologie des tumeurs et du développement
Signolle, Nicolas
Sauthier, Philippe
Pottier, Charles ; Université de Liège - ULiège > Master méd. (ex 4e master en 4 ans)
Gofflot, Stéphanie ; Centre Hospitalier Universitaire de Liège - CHU > Biothèque Ulg
De Cuypere, Marjolein ; Centre Hospitalier Universitaire de Liège - CHU > Service de gynécologie-obstétrique
Delvenne, Philippe ; Université de Liège > Département des sciences biomédicales et précliniques > Anatomie et cytologie pathologiques
Goffin, Frédéric ; Centre Hospitalier Universitaire de Liège - CHU > Service de gynécologie-obstétrique (CHR)
Noël, Agnès ✱; Université de Liège > Département des sciences cliniques > Labo de biologie des tumeurs et du développement
Kridelka, Frédéric ✱; Université de Liège > Département des sciences cliniques > Gynécologie-Obstétrique
Kaplan RN, Riba RD, Zacharoulis S, Bramley AH, Vincent L, Costa C, MacDonald DD, Jin DK, Shido K, Kerns SA, et al. VEGFR1-positive haematopoietic bone marrow progenitors initiate the pre-metastatic niche. Nature 2005; 438:820-7; PMID:16341007; http://dx.doi.org/10.1038/nature04186
Sceneay J, Smyth MJ, Moller A. The pre-metastatic niche:finding common ground. Cancer Metast Rev 2013; 32:449-64; PMID:23636348; http://dx.doi.org/10.1007/s10555-013-9420-1
Hirakawa S, Kodama S, Kunstfeld R, Kajiya K, Brown LF, Detmar M. VEGF-A induces tumor and sentinel lymph node lymphangiogenesis and promotes lymphatic metastasis. J Exp Med 2005; 201:1089-99; PMID:15809353; http://dx.doi.org/10.1084/jem.20041896
Qian CN, Berghuis B, Tsarfaty G, Bruch M, Kort EJ, Ditlev J, Tsarfaty I, Hudson E, Jackson DG, Petillo D, et al. Preparing the “soil”:the primary tumor induces vasculature reorganization in the sentinel lymph node before the arrival of metastatic cancer cells. Cancer Res 2006; 66:10365-76; PMID:17062557; http://dx.doi.org/10.1158/0008-5472.CAN-06-2977
Hirakawa S, Brown LF, Kodama S, Paavonen K, Alitalo K, Detmar M. VEGF-C-induced lymphangiogenesis in sentinel lymph nodes promotes tumor metastasis to distant sites. Blood 2007; 109:1010-7; PMID:17032920; http://dx.doi.org/10.1182/blood-2006-05-021758
Harrell MI, Iritani BM, Ruddell A. Tumor-induced sentinel lymph node lymphangiogenesis and increased lymph flow precede melanoma metastasis. Am J Pathol 2007; 170:774-86; PMID:17255343; http://dx.doi.org/10.2353/ajpath.2007.060761
Ozasa R, Ohno J, Iwahashi T, Taniguchi K. Tumor-induced lymphangiogenesis in cervical lymph nodes in oral melanoma-bearing mice. J Exp Clin Cancer Res 2012; 31:83; PMID:23031500; http://dx.doi.org/10.1186/1756-9966-31-83
Liersch R, Hirakawa S, Berdel WE, Mesters RM, Detmar M. Induced lymphatic sinus hyperplasia in sentinel lymph nodes by VEGF-C as the earliest premetastatic indicator. Int J Oncol 2012; 41:2073-8; PMID:23076721; http://dx.doi.org/10.3892/ijo.2012.1665
Ogawa F, Amano H, Eshima K, Ito Y, Matsui Y, Hosono K, Kitasato H, Iyoda A, Iwabuchi K, Kumagai Y, et al. Prostanoid induces premetastatic niche in regional lymph nodes. J Clin Investigat 2014; 124:4882-94; PMID:25271626; http://dx.doi.org/10.1172/JCI73530
Kawai H, Minamiya Y, Ito M, Saito H, Ogawa J. VEGF121 promotes lymphangiogenesis in the sentinel lymph nodes of non-small cell lung carcinoma patients. Lung Cancer 2008; 59:41-7; PMID:17868952; http://dx.doi.org/10.1016/j.lungcan.2007.08.001
Zhao YC, Ni XJ, Wang MH, Zha XM, Zhao Y, Wang S. Tumor-derived VEGF-C, but not VEGF-D, promotes sentinel lymph node lymphangiogenesis prior to metastasis in breast cancer patients. Med Oncol 2012; 29:2594-600; PMID:22562155; http://dx.doi.org/10.1007/s12032-012-0205-0
Ishii H, Chikamatsu K, Sakakura K, Miyata M, Furuya N, Masuyama K. Primary tumor induces sentinel lymph node lymphangiogenesis in oral squamous cell carcinoma. Oral Oncol 2010; 46:373-8; PMID:20308006; http://dx.doi.org/10.1016/j.oraloncology.2010.02.014
Chung MK, Do IG, Jung E, Son YI, Jeong HS, Baek CH. Lymphatic vessels and high endothelial venules are increased in the sentinel lymph nodes of patients with oral squamous cell carcinoma before the arrival of tumor cells. Ann Curg Oncol 2012; 19:1595-601; PMID:22124758; http://dx.doi.org/10.1245/s10434-011-2154-9
Wakisaka N, Hasegawa Y, Yoshimoto S, Miura K, Shiotani A, Yokoyama J, Sugasawa M, Moriyama-Kita M, Endo K, Yoshizaki T. Primary tumor-secreted lymphangiogenic factors induce pre-metastatic lymphvascular niche formation at sentinel lymph nodes in oral squamous cell carcinoma. PloS One 2015; 10:e0144056; PMID:26630663; http://dx.doi.org/10.1371/journal.pone.0144056
Garmy-Susini B, Avraamides CJ, Desgrosellier JS, Schmid MC, Foubert P, Ellies LG, Lowy AM, Blair SL, Vandenberg SR, Datnow B, et al. PI3Kalpha activates integrin alpha4beta1 to establish a metastatic niche in lymph nodes. Proc Natl Acad Sci USA 2013; 110:9042-7; PMID:23671068; http://dx.doi.org/10.1073/pnas.1219603110
Hirakawa S. From tumor lymphangiogenesis to lymphvascular niche. Cancer Sci 2009; 100:983-9; PMID:19385973; http://dx.doi.org/10.1111/j.1349-7006.2009.01142.x
Cochran AJ, Huang RR, Lee J, Itakura E, Leong SP, Essner R. Tumour-induced immune modulation of sentinel lymph nodes. Nat Rev Immunol 2006; 6:659-70; PMID:16932751; http://dx.doi.org/10.1038/nri1919
Nakamura S, Yaguchi T, Kawamura N, Kobayashi A, Sakurai T, Higuchi H, Takaishi H, Hibi T, Kawakami Y. TGF-beta1 in tumor microenvironments induces immunosuppression in the tumors and sentinel lymph nodes and promotes tumor progression. J Immunother 2014; 37:63-72; PMID:24509168; http://dx.doi.org/10.1097/CJI.0000000000000011
Ruddell A, Harrell MI, Furuya M, Kirschbaum SB, Iritani BM. B lymphocytes promote lymphogenous metastasis of lymphoma and melanoma. Neoplasia 2011; 13:748-57; PMID:21847366; http://dx.doi.org/10.1593/neo.11756
Ruddell A, Kelly-Spratt KS, Furuya M, Parghi SS, Kemp CJ. p19/Arf and p53 suppress sentinel lymph node lymphangiogenesis and carcinoma metastasis. Oncogene 2008; 27:3145-55; PMID:18059331; http://dx.doi.org/10.1038/sj.onc.1210973
Shrestha B, Hashiguchi T, Ito T, Miura N, Takenouchi K, Oyama Y, Kawahara K, Tancharoen S, Ki IY, Arimura N, et al. B cell-derived vascular endothelial growth factor A promotes lymphangiogenesis and high endothelial venule expansion in lymph nodes. J Immunol 2010; 184:4819-26; PMID:20308631; http://dx.doi.org/10.4049/jimmunol.0903063
Card CM, Yu SS, Swartz MA. Emerging roles of lymphatic endothelium in regulating adaptive immunity. J Clin Investigat 2014; 124:943-52; PMID:24590280; http://dx.doi.org/10.1172/JCI73316
Cohen JN, Guidi CJ, Tewalt EF, Qiao H, Rouhani SJ, Ruddell A, Farr AG, Tung KS, Engelhard VH. Lymph node-resident lymphatic endothelial cells mediate peripheral tolerance via Aire-independent direct antigen presentation. J Exp Med 2010; 207:681-8; PMID:20308365; http://dx.doi.org/10.1084/jem.20092465
Lund AW, Duraes FV, Hirosue S, Raghavan VR, Nembrini C, Thomas SN, Issa A, Hugues S, Swartz MA. VEGF-C promotes immune tolerance in B16 melanomas and cross-presentation of tumor antigen by lymph node lymphatics. Cell Rep 2012; 1:191-9; PMID:22832193; http://dx.doi.org/10.1016/j.celrep.2012.01.005
Tewalt EF, Cohen JN, Rouhani SJ, Guidi CJ, Qiao H, Fahl SP, Conaway MR, Bender TP, Tung KS, Vella AT, et al. Lymphatic endothelial cells induce tolerance via PD-L1 and lack of costimulation leading to high-level PD-1 expression on CD8 T cells. Blood 2012; 120:4772-82; PMID:22993390; http://dx.doi.org/10.1182/blood-2012-04-427013
Swartz MA, Lund AW. Lymphatic and interstitial flow in the tumour microenvironment:linking mechanobiology with immunity. Nat Rev Cancer 2012; 12:210-9; PMID:22362216; http://dx.doi.org/10.1038/nrc3186
Balsat C, Signolle N, Goffin F, Delbecque K, Plancoulaine B, Sauthier P, Samouelian V, Beliard A, Munaut C, Foidart JM, et al. Improved computer-assisted analysis of the global lymphatic network in human cervical tissues. Mod Pathol 2014; 27:887-98; PMID:24309324; http://dx.doi.org/10.1038/modpathol.2013.195
Riella LV, Paterson AM, Sharpe AH, Chandraker A. Role of the PD-1 pathway in the immune response. Am J Transplantat 2012; 12:2575-87; PMID:22900886; http://dx.doi.org/10.1111/j.1600-6143.2012.04224.x
De Silva NS, Klein U. Dynamics of B cells in germinal centres. Nat Rev Immunol 2015; 15:137-48; PMID:25656706; http://dx.doi.org/10.1038/nri3804
Pieper K, Grimbacher B, Eibel H. B-cell biology and development. J Allergy Clin Immunol 2013; 131:959-71; PMID:23465663; http://dx.doi.org/10.1016/j.jaci.2013.01.046
Agata Y, Kawasaki A, Nishimura H, Ishida Y, Tsubata T, Yagita H, Honjo T. Expression of the PD-1 antigen on the surface of stimulated mouse T and B lymphocytes. Int Immunol 1996; 8:765-72; PMID:8671665; http://dx.doi.org/10.1093/intimm/8.5.765
Nishimura H, Minato N, Nakano T, Honjo T. Immunological studies on PD-1 deficient mice:implication of PD-1 as a negative regulator for B cell responses. Int Immunol 1998; 10:1563-72; PMID:9796923; http://dx.doi.org/10.1093/intimm/10.10.1563
Good-Jacobson KL, Szumilas CG, Chen L, Sharpe AH, Tomayko MM, Shlomchik MJ. PD-1 regulates germinal center B cell survival and the formation and affinity of long-lived plasma cells. Nat Immunol 2010; 11:535-42; PMID:20453843; http://dx.doi.org/10.1038/ni.1877
Thibult ML, Mamessier E, Gertner-Dardenne J, Pastor S, Just-Landi S, Xerri L, Chetaille B, Olive D. PD-1 is a novel regulator of human B-cell activation. Int Immunol 2013; 25:129-37; PMID:23087177; http://dx.doi.org/10.1093/intimm/dxs098
Grotz TE, Jakub JW, Mansfield AS, Goldenstein R, Enninga EA, Nevala WK, Leontovich AA, Markovic SN. Evidence of Th2 polarization of the sentinel lymph node (SLN) in melanoma. Oncoimmunology 2015; 4:e1026504; PMID:26405583; http://dx.doi.org/10.1080/2162402X.2015.1026504
Fremd C, Schuetz F, Sohn C, Beckhove P, Domschke C. B cell-regulated immune responses in tumor models and cancer patients. Oncoimmunology 2013; 2:e25443; PMID:24073382; http://dx.doi.org/10.4161/onci.25443
Carmi Y, Spitzer MH, Linde IL, Burt BM, Prestwood TR, Perlman N, Davidson MG, Kenkel JA, Segal E, Pusapati GV, et al. Allogeneic IgG combined with dendritic cell stimuli induce antitumour T-cell immunity. Nature 2015; 521:99-104; PMID:25924063; http://dx.doi.org/10.1038/nature14424
Zirakzadeh AA, Marits P, Sherif A, Winqvist O. Multiplex B cell characterization in blood, lymph nodes, and tumors from patients with malignancies. J Immunol 2013; 190:5847-55; PMID:23630345; http://dx.doi.org/10.4049/jimmunol.1203279
Shalapour S, Font-Burgada J, Di Caro G, Zhong Z, Sanchez-Lopez E, Dhar D, Willimsky G, Ammirante M, Strasner A, Hansel DE, et al. Immunosuppressive plasma cells impede T-cell-dependent immunogenic chemotherapy. Nature 2015; 521:94-8; PMID:25924065; http://dx.doi.org/10.1038/nature14395
Ruddell A, Mezquita P, Brandvold KA, Farr A, Iritani BM. B lymphocyte-specific c-Myc expression stimulates early and functional expansion of the vasculature and lymphatics during lymphomagenesis. Am J Pathol 2003; 163:2233-45; PMID:14633598; http://dx.doi.org/10.1016/S0002-9440(10)63581-X
Roozendaal R, Mempel TR, Pitcher LA, Gonzalez SF, Verschoor A, Mebius RE, von Andrian UH, Carroll MC. Conduits mediate transport of low-molecular-weight antigen to lymph node follicles. Immunity 2009; 30:264-76; PMID:19185517; http://dx.doi.org/10.1016/j.immuni.2008.12.014
Christiansen AJ, Dieterich LC, Ohs I, Bachmann SB, Bianchi R, Proulx ST, Hollmen M, Aebischer D, Detmar M. Lymphatic endothelial cells attenuate inflammation via suppression of dendritic cell maturation. Oncotarget 2016; 7:39421-39435; PMID:27270646; http://dx.doi.org/10.18632/oncotarget.9820
