Abstract
Background: Reprogramming of immunosuppressive tumor-associated macrophages (TAMs) presents an attractive therapeutic strategy in cancer. The aim of this study was to explore the role of macrophage CD5L protein in TAM activity and assess its potential as a therapeutic target. Methods: Monoclonal antibodies (mAbs) against recombinant CD5L were raised by subcutaneous immunization of BALB/c mice. Peripheral blood monocytes were isolated from healthy donors and stimulated with IFN/LPS, IL4, IL10, and conditioned medium (CM) from different cancer cell lines in the presence of anti-CD5L mAb or controls. Subsequently, phenotypic markers, including CD5L, were quantified by flow cytometry, IF and RT-qPCR. Macrophage CD5L protein expression was studied in 55 human papillary lung adenocarcinoma (PAC) samples by IHC and IF. Anti-CD5L mAb and isotype control were administered intraperitoneally into a syngeneic Lewis Lung Carcinoma mouse model and tumor growth was measured. Tumor microenvironment (TME) changes were determined by flow cytometry, IHC, IF, Luminex, RNAseq and RT-qPCR. Findings: Cancer cell lines CM induced an immunosuppressive phenotype (increase in CD163, CD206, MERTK, VEGF and CD5L) in cultured macrophages. Accordingly, high TAM expression of CD5L in PAC was associated with poor patient outcome (Log-rank (Mantel–Cox) test p = 0.02). We raised a new anti-CD5L mAb that blocked the immunosuppressive phenotype of macrophages in vitro. Its administration in vivo inhibited tumor progression of lung cancer by altering the intratumoral myeloid cell population profile and CD4+ T-cell exhaustion phenotype, thereby significantly modifying the TME and increasing the inflammatory milieu. Interpretation: CD5L protein plays a key function in modulating the activity of macrophages and their interactions within the TME, which supports its role as a therapeutic target in cancer immunotherapy. Funding: For a full list of funding bodies, please see the Acknowledgements.
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | 104555 |
| Tidsskrift | EBioMedicine |
| Vol/bind | 91 |
| Antal sider | 22 |
| ISSN | 2352-3964 |
| DOI | |
| Status | Udgivet - maj 2023 |
Bibliografisk note
Funding Information:We thank Gerard Requena (Flow Cytometry Platform, IGTP) and M. Pilar Armengol (Translational Genomics Platform and Microscopy Platform, IGTP) for their valuable support in flow cytometry studies, and RNA and microscopy analysis, respectively. We also wish to acknowledge Maria Doladé Botías (Clinical Analyses and Biochemistry Dept. HUGTiP) for analyzing the mouse serum samples; Dr. Jo van Ginderachter (VIB, Belgium) for providing 3LL-R cells; Ramon Bartolí (IGTP-CIBERehd) for help writing the CMCIB ethical protocol for Animal Experimentation; Ana M Aransay for performing the RNAseq experiments (Genome Analysis Platform, CIC bioGUNE, Spain); Imane Chaib (Cancer Biology and Precision Medicine laboratory of Dr. Rosell ICO-IGTP) for providing the A459 and PC9 lung cancer cell lines used in the study; Elena Ramos for help with antibody purification (Dept. of Immunology and Oncology, CNB-CSIC) and Ismael Varela for his technical assistance with in vivo mice experiments. This study was supported by the Instituto de Salud Carlos III (ISCIII), and ERDFs from the EU, ‘Una manera de hacer Europa’, grants PI19/00523, 2019 PROD 00118 AGAUR, CI20-00152 and CC22-10181 Caixa Impulse to MRS, and from the Spanish Ministry of Science and Innovation grant PID2020-119875RB-I00 and PID2020-105404RB-I00 to AFV and LK, respectively. This research was also supported by the Consorcio Centro de Investigación Biomédica en Red (CIBERES and CIBEREHD). LSM, TP, JFD, and CM are supported by Juan de la Cierva (FJC2019-041213-I), PFIS (FI20/00115), FPI (PRE2018-085579), and Margarita Salas (MSGD2021-08) fellowships, respectively, all of which are programs run by the Spanish Government and NextGenerationEU, respectively. This study received funding from the EU's Horizon 2020 Research and Development Program under grant agreement nº 847762. The Innate Immunity lab and the UTE are accredited by the Catalan Agency for Management of University and Research Grants 2017-SGR-490/2021-SGR-01186 and 2017-SGR-500/2021 SGR 00920, respectively. MRS, CV, and CA are researchers at IGTP, which is a member of the CERCA network of institutes supported by the Health Department of the Government of Catalonia.
Funding Information:
This study was supported by the Instituto de Salud Carlos III (ISCIII), and ERDFs from the EU, ‘ Una manera de hacer Europa ’, grants PI19/00523 , 2019 PROD 00118 AGAUR , CI20-00152 and CC22-10181 Caixa Impulse to MRS, and from the Spanish Ministry of Science and Innovation grant PID2020-119875RB-I00 and PID2020-105404RB-I00 to AFV and LK, respectively. This research was also supported by the Consorcio Centro de Investigación Biomédica en Red (CIBERES and CIBEREHD). LSM, TP, JFD, and CM are supported by Juan de la Cierva (FJC2019-041213-I), PFIS (FI20/00115), FPI (PRE2018-085579), and Margarita Salas (MSGD2021-08) fellowships, respectively, all of which are programs run by the Spanish Government and NextGenerationEU, respectively. This study received funding from the EU’s Horizon 2020 Research and Development Program under grant agreement nº 847762. The Innate Immunity lab and the UTE are accredited by the Catalan Agency for Management of University and Research Grants 2017-SGR-490/2021-SGR-01186 and 2017-SGR-500/2021 SGR 00920, respectively. MRS, CV, and CA are researchers at IGTP, which is a member of the CERCA network of institutes supported by the Health Department of the Government of Catalonia.