Tumor cell invasion from the marginal sinus into extranodal veins during early-stage lymph node metastasis can be a starting point for hematogenous metastasis

Abstract

Aim: To investigate whether tumor cells in a lymph node (LN) can invade from the marginal sinus into extranodal veins via vessel branches that communicate with intranodal veins and whether this can be a starting point for hematogenous metastasis at the early stage of LN metastasis.

Methods: Vascular and lymphatic networks of LNs in MXH10/Mo-lpr/lpr mice were investigated using three-dimensional micro-computed tomography and histological methods. Flow in the blood vessel networks of LNs was investigated by fluorescence microscopy. Tumor cells were injected into the subiliac LNs of MXH10/Mo-lpr/lpr mice to induce metastasis to the proper axillary LNs. Tumor development in the proper axillary LN was detected using an in vivo bioluminescence imaging system. A two-dimensional image of the proper axillary LN microvasculature was reconstructed using a contrast-enhanced high-frequency ultrasound system.

Results: Extranodal veins communicated with intranodal veins via branches that penetrated the capsule, and blood flowed from intranodal veins to extranodal veins. Tumor cells that had metastasized to the marginal sinus invaded these communicating veins to develop hematogenous metastases.

Conclusion: Metastatic LNs that would be considered by clinical imaging to be stage N0 can be a starting point for hematogenous metastasis. The study findings highlight the need for the development of novel techniques for the diagnosis and treatment of early-stage LN metastasis, i.e., when standard diagnostic imaging might incorrectly classify the LN as stage N0.

Keywords

Lymph node / metastasis / N0 / lymph node-mediated hematogenous metastasis

References

[1] Starz H,Kramer KU,Wang H.A micromorphometry-based concept for routine classification of sentinel lymph node metastases and its clinical relevance for patients with melanoma..Cancer2001;91:2110-21

[2] Kelch ID,Sands GB,LeGrice IJ.Organ-wide 3D-imaging and topological analysis of the continuous microvascular network in a murine lymph node..Sci Rep2015;5:16534 PMCID:PMC4645097

[3] Jeong HS,Liao S,Cui CH.Investigation of the lack of angiogenesis in the formation of lymph node metastases..J Natl Cancer Inst2015;107:djv155 PMCID:PMC4651102

[4] Mikada M,Miura Y,Sakamoto M.Evaluation of the enhanced permeability and retention effect in the early stages of lymph node metastasis..Cancer Sci2017;108:846-52 PMCID:PMC5448659

[5] Qian CN,Tsarfaty G,Kort EJ.Preparing the “soil”: the primary tumor induces vasculature reorganization in the sentinel lymph node before the arrival of metastatic cancer cells..Cancer Res2006;66:10365-76

[6] Brown M,Leithner A,Schachner H.Lymph node blood vessels provide exit routes for metastatic tumor cell dissemination in mice..Science2018;359:1408-11

[7] Weaver DL,Krag DN,Anderson SJ.Effect of occult metastases on survival in node-negative breast cancer..N Engl J Med2011;364:412-21 PMCID:PMC3044504

[8] Shao L,Sakamoto M,Kodama T.Activation of latent metastases in the lung after resection of a metastatic lymph node in a lymph node metastasis mouse model..Biochem Biophys Res Commun2015;460:543-8

[9] Takeda K,Kodama T.Study of fluid dynamics reveals direct communications between lymphatic vessels and venous blood vessels at lymph nodes of mice..J Immunol Methods2017;445:1-9

[10] Shao L,Yagishita Y,Hatakeyama Y.Lymphatic mapping of mice with systemic lymphoproliferative disorder: usefulness as an inter-lymph node metastasis model of cancer..J Immunol Methods2013;389:69-78

[11] Nose M,Miyazaki T.Genomics of vasculitis: lessons from mouse models..Ann Vasc Dis2013;6:16-21 PMCID:PMC3634994

[12] Van den Broeck W,Simoens P.Anatomy and nomenclature of murine lymph nodes: descriptive study and nomenclatory standardization in BALB/cAnNCrl mice..J Immunol Methods2006;312:12-9

[13] Kochi T,Takeda A,Mori S.Characterization of the arterial anatomy of the murine hindlimb: functional role in the design and understanding of ischemia models..PLoS One2013;8:e84047 PMCID:PMC3875518

[14] Li L,Sakamoto M,Kodama T.Mouse model of lymph node metastasis via afferent lymphatic vessels for development of imaging modalities..PLoS One2013;8:e55797 PMCID:PMC3565997

[15] Miura Y,Ouchi T,Takeda K.Early diagnosis of lymph node metastasis: importance of intranodal pressures..Cancer Sci2016;107:224-32 PMCID:PMC4814246

[16] Shao L,Kato S,Kodama T.Communication between lymphatic and venous systems in mice..J Immunol Methods2015;424:100-5

[17] Kodama T,Kato S.Visualization of fluid drainage pathways in lymphatic vessels and lymph nodes using a mouse model to test a lymphatic drug delivery system..Biomed Opt Express2015;6:124-34 PMCID:PMC4317120

[18] Li XT,Tang L,Zhang XY.Evaluating local lymph node metastasis with magnetic resonance imaging, endoluminal ultrasound and computed tomography in rectal cancer: a meta-analysis..Colorectal Dis2015;17:O129-35

[19] Ito K,Yanagisawa Y,Mori S.High-accuracy ultrasound contrast agent detection method for diagnostic ultrasound imaging systems..Ultrasound Med Biol2015;41:3120-30

[20] Li L,Kodama M,Takahashi S.Enhanced sonographic imaging to diagnose lymph node metastasis: importance of blood vessel volume and density..Cancer Res2013;73:2082-92

[21] Sato T,Ouchi T,Sakamoto M.Monitoring of blood vessel density using contrast-enhanced high frequency ultrasound may facilitate early diagnosis of lymph node metastasis..J Cancer2017;8:704-15 PMCID:PMC5381158

[22] Iwaki T,Hayakawa T.A color atlas of sectional anatomy of the mouse.2015;TokyoMaruzen

[23] Fisher B.Transmigration of lymph nodes by tumor cells..Science1966;152:1397-8

[24] Fujii H,Takeda K,Kodama T.Optimal range of injection rates for a lymphatic drug delivery system..J Biophotonics2018;11:e201700401

[25] Kodama T,Tada A,Mori S.New concept for the prevention and treatment of metastatic lymph nodes using chemotherapy administered via the lymphatic network..Sci Rep2016;6:32506 PMCID:PMC5007471

[26] Tada A,Mori S.Therapeutic effect of cisplatin given with a lymphatic drug delivery system on false-negative metastatic lymph nodes..Cancer Sci2017;108:2115-21 PMCID:PMC5666029

[27] Morton DL,Cochran AJ,Nieweg OE.Final trial report of sentinel-node biopsy versus nodal observation in melanoma..N Engl J Med2014;370:599-609 PMCID:PMC4058881