Cancer cell – fibroblast crosstalk via HB-EGF/EGFR/MEK signaling promotes macrophage recruitment in squamous cell carcinoma

Interactions between cells in the tumor microenvironment (TME) shape cancer progression and patient outcomes. To gain new insights into how the TME influences cancer outcomes, we derive gene expression signatures indicative of signaling between stromal fibroblasts and cancer cells, and demonstrate their prognostic significance in multiple and independent squamous cell carcinoma cohorts. By leveraging information within the signatures, we discover that the HB-EGF/EGFR/MEK axis represents a hub of tumor – stroma crosstalk, promoting the expression of CSF2 and LIF and favoring the recruitment of macrophages. Together these analyses demonstrate the utility of our approach for interrogating the extent and consequences of TME crosstalk. ### Competing Interest Statement G.G. is funded by Merck Sharp & Dohme Corp, New Jersey, USA (LKR190557). A.R. acknowledges funding from the Spanish Society for Medical Oncology (Beca Fundacion SEOM), a CRUK accelerator grant to The Francis Crick Institute, and the Royal Marsden NIHR/BRC Bridge funding program. E.H. is supported by the Japan Society for the Promotion of Science (JSPS) Kakenhi Grant [20H03510]. Y.N was supported by a Grant-in-aid for JSPS Overseas Research Fellowship (No. 201860634). K.H. and A.M. acknowledge funding by the Wellcome Trust, ICR/RM NIHR Biomedical Research Centre, The Institute of Cancer Research/Royal Marsden Hospital Centre for Translational Immunotherapy, CRUK Head and Neck Programme Grant (C7224/A23275) and ICR/RM CRUK RadNet Centre of Excellence (C7224/A28724). Disclosures: Honoraria: Arch Oncology (Inst), AstraZeneca (Inst), BMS (Inst), Boehringer Ingelheim (Inst), Codiak Biosciences (Inst), F-Star Therapeutics (Inst), Inzen Therapeutics (Inst), Merck Serono (Inst), MSD (Inst), Oncolys Biopharma (Inst), Pfizer (Inst), Replimune (Inst), VacV Biotherapeutics (Inst); Consulting or Advisory Role: Arch Oncology (Inst), AstraZeneca (Inst), BMS (Inst), Boehringer Ingelheim (Inst), Inzen Therapeutics (Inst), Merck Serono (Inst), MSD (Inst), Oncolys BioPharma (Inst), Replimune (Inst); Speakers Bureau: BMS (Inst), Merck Serono (Inst), MSD (Inst); Research Funding: AstraZeneca (Inst), Boehringer Ingelheim (Inst), Merck Sharp & Dohme (Inst), Replimune (Inst). S.B. declares no conflict of Interest and funding support from ICR/RM NIHR Biomedical Research Centre, CRUK Head and Neck Programme Grant (C7224/A23275), ICR/RM CRUK RadNet Centre of Excellence (C7224/A28724) and the Medical Research Council Developmental Pathway Funding Scheme [MR/R015589/1]. D.B. was supported by funding from a Cancer Research UK (CRUK) Early Detection and Diagnosis Project award, the Idea to Innovation (i2i) Crick translation scheme supported by the Medical Research Council, the National Institute for Health Research Biomedical Research Centre and the Breast Cancer Research Foundation (BCRF). D.B. reports personal fees from NanoString and AstraZeneca, and has a patent PCT/GB2020/050221 issued on methods for cancer prognostication. C.S. acknowledges grant support from AstraZeneca, Boehringer-Ingelheim, Bristol Myers Squibb, Pfizer, Roche-Ventana, Invitae (previously Archer Dx Inc collaboration in minimal residual disease sequencing technologies), and Ono Pharmaceutical. He is an AstraZeneca Advisory Board member and Chief Investigator for the AZ MeRmaiD 1 and 2 clinical trials and is also Co-Chief Investigator of the NHS Galleri trial funded by GRAIL and a paid member of GRAIL Scientific Advisory Board. He receives consultant fees from Achilles Therapeutics (also SAB member), Bicycle Therapeutics (also a SAB member), Genentech, Medicxi, Roche Innovation Centre Shanghai, Metabomed (until July 2022), and the Sarah Cannon Research Institute. C.S has received honoraria from Amgen, AstraZeneca, Pfizer, Novartis, GlaxoSmithKline, MSD, Bristol Myers Squibb, Illumina, and Roche-Ventana. C.S. had stock options in Apogen Biotechnologies and GRAIL until June 2021, and currently has stock options in Epic Bioscience, Bicycle Therapeutics, and has stock options and is co-founder of Achilles Therapeutics. C.S. holds patents relating to assay technology to detect tumor recurrence (PCT/GB2017/053289); to targeting neoantigens (PCT/EP2016/059401), identifying patent response to immune checkpoint blockade (PCT/EP2016/071471), determining HLA LOH (PCT/GB2018/052004), predicting survival rates of patients with cancer (PCT/GB2020/050221), identifying patients who respond to cancer treatment (PCT/GB2018/051912), US patent relating to detecting tumor mutations (PCT/US2017/28013), methods for lung cancer detection (US20190106751A1) and both a European and US patent related to identifying insertion/deletion mutation targets (PCT/GB2018/051892). C.S. is a Royal Society Napier Research Professor (RSRP\R\210001). This work was supported by the Francis Crick Institute that receives its core funding from Cancer Research UK (CC2041), the UK Medical Research Council (CC2041), and the Wellcome Trust (CC2041). For the purpose of Open Access, the author has applied a CC BY public copyright licence to any Author Accepted Manuscript version arising from this submission. C.S. is funded by Cancer Research UK (TRACERx (C11496/A17786), PEACE (C416/A21999) and CRUK Cancer Immunotherapy Catalyst Network); Cancer Research UK Lung Cancer Centre of Excellence (C11496/A30025); the Rosetrees Trust, Butterfield and Stoneygate Trusts; NovoNordisk Foundation (ID16584); Royal Society Professorship Enhancement Award (RP/EA/180007); National Institute for Health Research (NIHR) University College London Hospitals Biomedical Research Centre; the Cancer Research UK-University College London Centre; Experimental Cancer Medicine Centre; the Breast Cancer Research Foundation (US) BCRF 22 157; Cancer Research UK Early Detection an Diagnosis Primer Award (Grant EDDPMA Nov21/100034); and The Mark Foundation for Cancer Research Aspire Award (Grant 21 029 ASP). This work was supported by a Stand Up To Cancer‐LUNGevity-American Lung Association Lung Cancer Interception Dream Team Translational Research Grant (Grant Number: SU2C AACR DT23 17 to S.M. Dubinett and A.E. Spira). Stand Up To Cancer is a division of the Entertainment Industry Foundation. Research grants are administered by the American Association for Cancer Research, the Scientific Partner of SU2C. C.S. is in receipt of an ERC Advanced Grant (PROTEUS) from the European Research Council under the European Union Horizon 2020 research and innovation programme (grant agreement no. 835297). E.S. is supported by the Francis Crick Institute which receives its core funding from Cancer Research UK (CC2040), the UK Medical Research Council (CC2040), and the Wellcome Trust (CC2040). E.S. is additionally funded by the European Research Council ERCAdG CAN\_ORGANISE 101019366. E.S. declares research funding from Merck Sharp & Dohme and Astrazeneca and consultancy work for Phenomic AI and Theolytics. For the purpose of Open Access, the author has applied a CC BY public copyright licence to any Author Accepted Manuscript version arising from this submission. RNA sequencing of the UK\_HPV positive cohort was funded by the NIHR through the RMH BRC Pump Priming theme (grant B140). The INOVATE study was funded by the Medical Research Council Developmental Pathway Funding Scheme [MR/R015589/1]. The INSIGHT2 trial was supported by CRUK Head and Neck Programme Grant (C7224/A23275). This study represents independent research supported by the National Institute for Health Research (NIHR) Biomedical Research Centre at the Royal Marsden National Health Service Foundation Trust and the Institute of Cancer Research, London, UK.