American Association for Cancer Research
00085472can160598-sup-162803_2_supp_3775814_lhw5sx.pdf (3.25 MB)

Supplementary Figures & Tables from RORγt+ Innate Lymphoid Cells Promote Lymph Node Metastasis of Breast Cancers

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journal contribution
posted on 2023-03-31, 01:10 authored by Sheeba Irshad, Fabian Flores-Borja, Katherine Lawler, James Monypenny, Rachel Evans, Victoria Male, Peter Gordon, Anthony Cheung, Patrycja Gazinska, Farzana Noor, Felix Wong, Anita Grigoriadis, Gilbert O. Fruhwirth, Paul R. Barber, Natalie Woodman, Dominic Patel, Manuel Rodriguez-Justo, Julie Owen, Stewart G. Martin, Sarah E. Pinder, Cheryl E. Gillett, Simon P. Poland, Simon Ameer-Beg, Frank McCaughan, Leo M. Carlin, Uzma Hasan, David R. Withers, Peter Lane, Borivoj Vojnovic, Sergio A. Quezada, Paul Ellis, Andrew N.J. Tutt, Tony Ng

Figure S1, Related to Figure 1 FACS and immunofluorescence validation of Lin-CD127+CD90.2+NKP46- gated ILC3 cells Figure S2, Related to Figure 2 ELISA quantification of CXCL13 and CCL21 chemokines levels in conditioned media (CM) obtained from the bone marrow derived MSC cell line (HS-5) and from the breast cancer cell line 4T1.2 cell line is shown. Figure S3, Related to Figure 2 Proliferation assay of ILC3 cells and ELISA quantification of CXCL13 and CCL21 from siRNA transfected MSC cells. Figure S4, Related to Figure 4 CXCR5 expression on MSC cells. Figure S5, Related to Figure 5 Identification of a RORγT+CD127+CD3- ILC3 cells within the tertiary lymphoid structures within breast cancers. Figure S6, Related to Figure 5 Expression of lymphoid chemokine and chemokine receptor genes in breast cancer datasets. Table S1, Related to Figure 5 Table S1: Clinico-pathological characteristics for the METABRIC sample.



Cancer cells tend to metastasize first to tumor-draining lymph nodes, but the mechanisms mediating cancer cell invasion into the lymphatic vasculature remain little understood. Here, we show that in the human breast tumor microenvironment (TME), the presence of increased numbers of RORγt+ group 3 innate lymphoid cells (ILC3) correlates with an increased likelihood of lymph node metastasis. In a preclinical mouse model of breast cancer, CCL21-mediated recruitment of ILC3 to tumors stimulated the production of the CXCL13 by TME stromal cells, which in turn promoted ILC3–stromal interactions and production of the cancer cell motile factor RANKL. Depleting ILC3 or neutralizing CCL21, CXCL13, or RANKL was sufficient to decrease lymph node metastasis. Our findings establish a role for RORγt+ILC3 in promoting lymphatic metastasis by modulating the local chemokine milieu of cancer cells in the TME. Cancer Res; 77(5); 1083–96. ©2017 AACR.