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Heterotypic interactions between stromal, immune and malignant epithelial cells play important roles in solid tumour progression and therapeutic response. Such insights present promising candidates to further investigate for new therapeutic strategies in the treatment of TNBCs. Importantly, the investigation of gene signatures from inflammatory-CAFs and differentiated-PVL cells in independent TNBC patient cohorts revealed strong associations with cytotoxic T-cell dysfunction and exclusion, respectively. Using cell signalling predictions, we provide evidence that stromal-immune crosstalk acts via a diverse array of immunoregulatory molecules.

We showed that these stromal states have distinct morphologies, spatial relationships and functional properties in regulating the extracellular matrix. PVL cells clustered into two states consistent with a differentiated and immature phenotype. CAFs clustered into two states: the first with features of myofibroblasts and the second characterised by high expression of growth factors and immunomodulatory molecules. Single-cell RNA sequencing of five TNBCs revealed two cancer-associated fibroblast (CAF) and two perivascular-like (PVL) subpopulations. Stromal heterogeneity in human triple-negative breast cancers (TNBCs) remains poorly understood, limiting the development of stromal-targeted therapies. The tumour stroma regulates nearly all stages of carcinogenesis. 16 Australian Clinical Laboratories, Northern Beaches Hospital, Frenchs Forest, NSW, Australia.15 UNSW Cellular Genomics Futures Institute, University of New South Wales, Sydney, NSW, Australia.14 RIKEN Center for Integrative Medical Sciences, Yokohama, Japan.13 Royal Prince Alfred Institute of Academic Surgery, Sydney University, Sydney, NSW, Australia.12 Department of Breast Surgery, Chris O'Brien Lifehouse, Camperdown, NSW, Australia.

11 Southside Clinical Unit, Faculty of Medicine, University of Queensland, Brisbane, Qld, Australia.10 Pathology Queensland, Princess Alexandra Hospital, Brisbane, Qld, Australia.9 Royal Prince Alfred Hospital, Camperdown, NSW, Australia.8 St Vincent's Hospital, Darlinghurst, NSW, Australia.7 The Strathfield Breast Centre, Strathfield, NSW, Australia.6 Chris O'Brien Lifehouse, Camperdown, NSW, Australia.5 Garvan-Weizmann Centre for Cellular Genomics, Garvan Institute of Medical Research, Sydney, NSW, Australia.4 Harry Perkins Institute of Medical Research, QEII Medical Centre and Centre for Medical Research, The University of Western Australia, Nedlands, Perth, WA, Australia.Chan School of Public Health, Boston, MA, USA 3 Department of Data Sciences, Center for Functional Cancer Epigenetics, Dana-Farber Cancer Institute, Harvard T.H.2 St Vincent's Clinical School, Faculty of Medicine, UNSW Sydney, Sydney, NSW, Australia.

1 The Kinghorn Cancer Centre and Cancer Research Division, Garvan Institute of Medical Research, Darlinghurst, NSW, Australia.

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