Tumor-Infiltrating B Lymphocyte Profiling Identifies IgG-Biased, Clonally Expanded Prognostic Phenotypes In Triple-Negative Breast Cancer

TY - JOUR

T1 - Tumor-Infiltrating B Lymphocyte Profiling Identifies IgG-Biased, Clonally Expanded Prognostic Phenotypes In Triple-Negative Breast Cancer

AU - Harris, Robert J.

AU - Cheung, Anthony

AU - Ng, Joseph C.F.

AU - Laddach, Roman

AU - Chenoweth, Alicia M.

AU - Crescioli, Silvia

AU - Fittall, Matthew

AU - Dominguez-Rodriguez, Diana

AU - Roberts, James

AU - Levi, Dina

AU - Liu, Fangfang

AU - Alberts, Elena

AU - Quist, Jelmar

AU - Santaolalla, Aida

AU - Pinder, Sarah E.

AU - Gillett, Cheryl

AU - Hammar, Niklas

AU - Irshad, Sheeba

AU - van Hemelrijck, Mieke

AU - Dunn-Walters, Deborah K.

AU - Fraternali, Franca

AU - Spicer, James F.

AU - Lacy, Katie E.

AU - Tsoka, Sophia

AU - Grigoriadis, Anita

AU - Tutt, Andrew N.J.

AU - Karagiannis, Sophia N.

N1 - Funding Information: The authors acknowledge support by Breast Cancer Now (147; KCL-BCN-Q3); the Cancer Research UK King’s Health Partners Center at King’s College London (C604/A25135); Cancer Research UK (C30122/A11527; C30122/A15774); the Medical Research Council (MR/L023091/1); CR UK//NIHR in England/DoH for Scotland, Wales and Northern Ireland Experimental Cancer Medicine Center (C10355/A15587). The research was supported by the National Institute for Health Research Biomedical Research Center based at Guy’s and St Thomas’ NHS Foundation Trust and King’s College London (IS-BRC-1215-20006). The authors are solely responsible for study design, data collection, analysis, decision to publish, and preparation of the manuscript. The views expressed are those of Funding Information: D.K. Dunn-Walters reports grants from MRC during the conduct of the study. A.N.J. Tutt reports grants from Breast Cancer Now Charity and CRUK during the conduct of the study as well as reports personal fees from Inbiomo-tion, CRUK, MD Anderson, and reports other support from Tesaro/ GlaxoSmithKline, AstraZeneca, AstraZeneca, Merck KGAA, as well as personal fees from Pfizer, Vertex, Artios, Prime Oncology, and reports other support from Medivation, Myriad Genetics, and reports personal fees from Gilead, other support from AstraZeneca, and grants from AstraZeneca outside the submitted work; as well as a patent for AstraZeneca with royalties paid to the Institute of Cancer Research with royalties paid from AstraZeneca. S.N. Karagiannis reports grants from Breast Cancer Now, Cancer Research UK, Medical Research Council, and National Institute for Health Research during the conduct of the study as well as reports grants from Epsilogen Ltd. outside the submitted work; and patents on novel antibodies for cancer therapy. No disclosures were reported by the other authors. Publisher Copyright: © 2021 The Authors; Published by the American Association for Cancer Research Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

PY - 2021/8/15

Y1 - 2021/8/15

N2 - In breast cancer, humoral immune responses may contribute to clinical outcomes, especially in more immunogenic subtypes. Here we investigated B lymphocyte subsets, immunoglobulin expression, and clonal features in breast tumors, focusing on aggressive triple-negative breast cancers (TNBC). In samples from TNBC patients and healthy volunteers, circulating and tumor-infiltrating B lymphocyte (TIL-B) were evaluated. CD20+CD27+IgD- isotype-switched B lymphocytes were increased in tumors, compared with matched blood. TIL-B frequently formed stromal clusters with T lymphocytes andengaged in bidirectional functional crosstalk, consistent with gene signatures associated with lymphoid assembly, co-stimulation, cytokine-cytokine receptor interactions, cytotoxic T cell activation, and T cell-dependent B cell activation. TIL-B upregulated B cell receptor (BCR) pathway molecules FOS and JUN, germinal center chemokine regulator RGS1, activation marker CD69, and TNFα signal transduction via NFκB, suggesting BCR-immune complex formation. Expression of genes associated with B lymphocyte recruitment and lymphoid assembly, including CXCL13, CXCR4, DC-LAMP, was elevated in TNBC compared with other subtypes and normal breast. TIL-B-rich tumors showed expansion ofIgG but not IgA isotypes, and IgG isotype-switching positively associated with survival outcomes in TNBC. Clonal expansion was biased towards IgG, showing expansive clonal families with specific variable region gene combinations and narrow repertoires. Stronger positive selection pressure was present in the complementary determining regions (CDRs) of IgG compared to their clonally related IgA in tumor samples. Overall, class-switched B lymphocyte lineage traits were conspicuous in TNBC, associated with improved clinical outcomes, and conferred IgG-biased, clonally expanded, and likely antigen-driven humoralresponses.

AB - In breast cancer, humoral immune responses may contribute to clinical outcomes, especially in more immunogenic subtypes. Here we investigated B lymphocyte subsets, immunoglobulin expression, and clonal features in breast tumors, focusing on aggressive triple-negative breast cancers (TNBC). In samples from TNBC patients and healthy volunteers, circulating and tumor-infiltrating B lymphocyte (TIL-B) were evaluated. CD20+CD27+IgD- isotype-switched B lymphocytes were increased in tumors, compared with matched blood. TIL-B frequently formed stromal clusters with T lymphocytes andengaged in bidirectional functional crosstalk, consistent with gene signatures associated with lymphoid assembly, co-stimulation, cytokine-cytokine receptor interactions, cytotoxic T cell activation, and T cell-dependent B cell activation. TIL-B upregulated B cell receptor (BCR) pathway molecules FOS and JUN, germinal center chemokine regulator RGS1, activation marker CD69, and TNFα signal transduction via NFκB, suggesting BCR-immune complex formation. Expression of genes associated with B lymphocyte recruitment and lymphoid assembly, including CXCL13, CXCR4, DC-LAMP, was elevated in TNBC compared with other subtypes and normal breast. TIL-B-rich tumors showed expansion ofIgG but not IgA isotypes, and IgG isotype-switching positively associated with survival outcomes in TNBC. Clonal expansion was biased towards IgG, showing expansive clonal families with specific variable region gene combinations and narrow repertoires. Stronger positive selection pressure was present in the complementary determining regions (CDRs) of IgG compared to their clonally related IgA in tumor samples. Overall, class-switched B lymphocyte lineage traits were conspicuous in TNBC, associated with improved clinical outcomes, and conferred IgG-biased, clonally expanded, and likely antigen-driven humoralresponses.

KW - B lymphocyte

KW - Memory B lymphocyte

KW - IgG isotype-switching

KW - Breast Cancer

KW - Triple-negative breast cancer

KW - Tumor-infiltrating lymphocytes

KW - Immunoglobulin repertoire

KW - Tumor Microenvironment/immunology

KW - Clonal expansion

UR - http://www.scopus.com/inward/record.url?scp=85112812647&partnerID=8YFLogxK

U2 - 10.1158/0008-5472.CAN-20-3773

DO - 10.1158/0008-5472.CAN-20-3773

M3 - Article

C2 - 34224371

AN - SCOPUS:85112812647

SN - 0008-5472

VL - 81

SP - 4290

EP - 4304

JO - Cancer Research

JF - Cancer Research

IS - 16

ER -