Exploitable mechanisms of antibody and CAR mediated macrophage cytotoxicity

Abstract Macrophages infiltrate solid tumors and either support survival or induce cancer cell death through phagocytosis or cytotoxicity. To uncover regulators of macrophage cytotoxicity towards cancer cells, we perform two co-culture CRISPR screens using CAR-macrophages targeting different tumor a...

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Main Authors: Tianyi Liu, Meng Zhang, Tatyanah Farsh, Haolong Li, Audrey Kishishita, Abhilash Barpanda, Stanley G. Leung, Jun Zhu, Hyuncheol Jung, Junjie Tony Hua, Xiaolin Zhu, Alexander B. Kim, Young Ah Goo, Minsoo Son, Jaenyeon Kim, Aish Subramanian, Martin Sjöström, Katherine C. Fuh, Jocelyn S. Chapman, Julia Carnevale, Luke A. Gilbert, Aparna Lakkaraju, Peter M. Bruno, David Quigley, Arun P. Wiita, Felix Y. Feng, Carl J. DeSelm
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-60745-x
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author Tianyi Liu
Meng Zhang
Tatyanah Farsh
Haolong Li
Audrey Kishishita
Abhilash Barpanda
Stanley G. Leung
Jun Zhu
Hyuncheol Jung
Junjie Tony Hua
Xiaolin Zhu
Alexander B. Kim
Young Ah Goo
Minsoo Son
Jaenyeon Kim
Aish Subramanian
Martin Sjöström
Katherine C. Fuh
Jocelyn S. Chapman
Julia Carnevale
Luke A. Gilbert
Aparna Lakkaraju
Peter M. Bruno
David Quigley
Arun P. Wiita
Felix Y. Feng
Carl J. DeSelm
author_facet Tianyi Liu
Meng Zhang
Tatyanah Farsh
Haolong Li
Audrey Kishishita
Abhilash Barpanda
Stanley G. Leung
Jun Zhu
Hyuncheol Jung
Junjie Tony Hua
Xiaolin Zhu
Alexander B. Kim
Young Ah Goo
Minsoo Son
Jaenyeon Kim
Aish Subramanian
Martin Sjöström
Katherine C. Fuh
Jocelyn S. Chapman
Julia Carnevale
Luke A. Gilbert
Aparna Lakkaraju
Peter M. Bruno
David Quigley
Arun P. Wiita
Felix Y. Feng
Carl J. DeSelm
author_sort Tianyi Liu
collection DOAJ
description Abstract Macrophages infiltrate solid tumors and either support survival or induce cancer cell death through phagocytosis or cytotoxicity. To uncover regulators of macrophage cytotoxicity towards cancer cells, we perform two co-culture CRISPR screens using CAR-macrophages targeting different tumor associated antigens. Both identify ATG9A as an important regulator of this cytotoxic activity. In vitro and in vivo, ATG9A depletion in cancer cells sensitizes them to macrophage-mediated killing. Proteomic and lipidomic analyses reveal that ATG9A deficiency impairs the cancer cell response to macrophage-induced plasma membrane damage through defective lysosomal exocytosis, reduced ceramide production, and disrupted caveolar endocytosis. Depleting non-cytotoxic macrophages using CSF1R inhibition while preventing ATG9A-mediated tumor membrane repair enhances the anti-tumor activity of therapeutic antibodies in mice. Thus, macrophage cytotoxicity plays an important role in tumor elimination during antibody or CAR-macrophage treatment, and inhibiting tumor membrane repair via ATG9A, particularly in combination with cytotoxic macrophage enrichment through CSF1R inhibition, improves tumor-targeting macrophage efficacy.
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spelling doaj-art-23c81c9621ef4e5f88e89d09abffe8c52025-08-20T03:03:33ZengNature PortfolioNature Communications2041-17232025-07-0116112210.1038/s41467-025-60745-xExploitable mechanisms of antibody and CAR mediated macrophage cytotoxicityTianyi Liu0Meng Zhang1Tatyanah Farsh2Haolong Li3Audrey Kishishita4Abhilash Barpanda5Stanley G. Leung6Jun Zhu7Hyuncheol Jung8Junjie Tony Hua9Xiaolin Zhu10Alexander B. Kim11Young Ah Goo12Minsoo Son13Jaenyeon Kim14Aish Subramanian15Martin Sjöström16Katherine C. Fuh17Jocelyn S. Chapman18Julia Carnevale19Luke A. Gilbert20Aparna Lakkaraju21Peter M. Bruno22David Quigley23Arun P. Wiita24Felix Y. Feng25Carl J. DeSelm26University of California, San Francisco, Helen Diller Family Comprehensive Cancer CenterUniversity of California, San Francisco, Helen Diller Family Comprehensive Cancer CenterUniversity of California, San Francisco, Helen Diller Family Comprehensive Cancer CenterUniversity of California, San Francisco, Helen Diller Family Comprehensive Cancer CenterDepartment of Pharmaceutical Chemistry, University of California, San FranciscoDepartment of Laboratory Medicine, University of California, San FranciscoUniversity of California, San Francisco, Helen Diller Family Comprehensive Cancer CenterUniversity of California, San Francisco, Helen Diller Family Comprehensive Cancer CenterGladstone–UCSF Institute of Genomic ImmunologyUniversity of California, San Francisco, Helen Diller Family Comprehensive Cancer CenterUniversity of California, San Francisco, Helen Diller Family Comprehensive Cancer CenterDepartment of Radiation Oncology, Washington University School of MedicineMass Spectrometry Technology Access Center at McDonnell Genome Institute (MTAC@MGI) at Washington University School of MedicineMass Spectrometry Technology Access Center at McDonnell Genome Institute (MTAC@MGI) at Washington University School of MedicineMass Spectrometry Technology Access Center at McDonnell Genome Institute (MTAC@MGI) at Washington University School of MedicineUniversity of California, San Francisco, Helen Diller Family Comprehensive Cancer CenterDepartment of Radiation Oncology, University of California, San FranciscoUniversity of California, San Francisco, Helen Diller Family Comprehensive Cancer CenterUniversity of California, San Francisco, Helen Diller Family Comprehensive Cancer CenterUniversity of California, San Francisco, Helen Diller Family Comprehensive Cancer CenterUniversity of California, San Francisco, Helen Diller Family Comprehensive Cancer CenterPharmaceutical Sciences and Pharmacogenomics, University of California San FranciscoUniversity of California, San Francisco, Helen Diller Family Comprehensive Cancer CenterUniversity of California, San Francisco, Helen Diller Family Comprehensive Cancer CenterDepartment of Laboratory Medicine, University of California, San FranciscoUniversity of California, San Francisco, Helen Diller Family Comprehensive Cancer CenterDepartment of Radiation Oncology, Washington University School of MedicineAbstract Macrophages infiltrate solid tumors and either support survival or induce cancer cell death through phagocytosis or cytotoxicity. To uncover regulators of macrophage cytotoxicity towards cancer cells, we perform two co-culture CRISPR screens using CAR-macrophages targeting different tumor associated antigens. Both identify ATG9A as an important regulator of this cytotoxic activity. In vitro and in vivo, ATG9A depletion in cancer cells sensitizes them to macrophage-mediated killing. Proteomic and lipidomic analyses reveal that ATG9A deficiency impairs the cancer cell response to macrophage-induced plasma membrane damage through defective lysosomal exocytosis, reduced ceramide production, and disrupted caveolar endocytosis. Depleting non-cytotoxic macrophages using CSF1R inhibition while preventing ATG9A-mediated tumor membrane repair enhances the anti-tumor activity of therapeutic antibodies in mice. Thus, macrophage cytotoxicity plays an important role in tumor elimination during antibody or CAR-macrophage treatment, and inhibiting tumor membrane repair via ATG9A, particularly in combination with cytotoxic macrophage enrichment through CSF1R inhibition, improves tumor-targeting macrophage efficacy.https://doi.org/10.1038/s41467-025-60745-x
spellingShingle Tianyi Liu
Meng Zhang
Tatyanah Farsh
Haolong Li
Audrey Kishishita
Abhilash Barpanda
Stanley G. Leung
Jun Zhu
Hyuncheol Jung
Junjie Tony Hua
Xiaolin Zhu
Alexander B. Kim
Young Ah Goo
Minsoo Son
Jaenyeon Kim
Aish Subramanian
Martin Sjöström
Katherine C. Fuh
Jocelyn S. Chapman
Julia Carnevale
Luke A. Gilbert
Aparna Lakkaraju
Peter M. Bruno
David Quigley
Arun P. Wiita
Felix Y. Feng
Carl J. DeSelm
Exploitable mechanisms of antibody and CAR mediated macrophage cytotoxicity
Nature Communications
title Exploitable mechanisms of antibody and CAR mediated macrophage cytotoxicity
title_full Exploitable mechanisms of antibody and CAR mediated macrophage cytotoxicity
title_fullStr Exploitable mechanisms of antibody and CAR mediated macrophage cytotoxicity
title_full_unstemmed Exploitable mechanisms of antibody and CAR mediated macrophage cytotoxicity
title_short Exploitable mechanisms of antibody and CAR mediated macrophage cytotoxicity
title_sort exploitable mechanisms of antibody and car mediated macrophage cytotoxicity
url https://doi.org/10.1038/s41467-025-60745-x
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