Cell therapy using ex vivo reprogrammed macrophages enhances antitumor immune responses in melanoma

Satish Kumar Noonepalle, Lombardi Comprehensive Cancer Center, Georgetown University, 3970 Reservoir Road, NW, E416 Research Bldg, Washington, DC, 20057, USA.
Maria Gracia-Hernandez, The George Washington University, Washington, DC, USA.
Nima Aghdam, Lombardi Comprehensive Cancer Center, Georgetown University, 3970 Reservoir Road, NW, E416 Research Bldg, Washington, DC, 20057, USA.
Michael Berrigan, The George Washington University, Washington, DC, USA.
Hawa Coulibaly, Lombardi Comprehensive Cancer Center, Georgetown University, 3970 Reservoir Road, NW, E416 Research Bldg, Washington, DC, 20057, USA.
Xintang Li, Lombardi Comprehensive Cancer Center, Georgetown University, 3970 Reservoir Road, NW, E416 Research Bldg, Washington, DC, 20057, USA.
Christian Zevallos-Delgado, The George Washington University, Washington, DC, USA.
Andrew Pletcher, The George Washington University, Washington, DC, USA.
Bryan Weselman, Lombardi Comprehensive Cancer Center, Georgetown University, 3970 Reservoir Road, NW, E416 Research Bldg, Washington, DC, 20057, USA.
Erica Palmer, The George Washington University, Washington, DC, USA.
Tessa Knox, The George Washington University, Washington, DC, USA.
Eduardo Sotomayor, Tampa General Hospital, Tampa, FL, USA.
Katherine B. Chiappinelli, The George Washington University, Washington, DC, USA.
Duncan Wardrop, University of Illinois at Chicago, Chicago, IL, USA.
Anelia Horvath, The George Washington University, Washington, DC, USA.
Brett A. Shook, The George Washington University, Washington, DC, USA.
Norman Lee, The George Washington University, Washington, DC, USA.
Anatoly Dritschilo, Lombardi Comprehensive Cancer Center, Georgetown University, 3970 Reservoir Road, NW, E416 Research Bldg, Washington, DC, 20057, USA.
Rohan Fernandes, The George Washington University, Washington, DC, USA.
Karthik Musunuri, Avstera Therapeutics, Malvern, PA, USA.
Maho Shibata, The George Washington University, Washington, DC, USA.
Alejandro Villagra, Lombardi Comprehensive Cancer Center, Georgetown University, 3970 Reservoir Road, NW, E416 Research Bldg, Washington, DC, 20057, USA. Alejandro.villagra@georgetown.edu.

Document Type

Journal Article

Publication Date

9-14-2024

Journal

Journal of experimental & clinical cancer research : CR

Volume

43

Issue

1

DOI

10.1186/s13046-024-03182-w

Abstract

BACKGROUND: Macrophage-based cell therapies have shown modest success in clinical trials, which can be attributed to their phenotypic plasticity, where transplanted macrophages get reprogrammed towards a pro-tumor phenotype. In most tumor types, including melanoma, the balance between antitumor M1-like and tumor-promoting M2-like macrophages is critical in defining the local immune response with a higher M1/M2 ratio favoring antitumor immunity. Therefore, designing novel strategies to increase the M1/M2 ratio in the TME has high clinical significance and benefits macrophage-based cell therapies. METHODS: In this study, we reprogrammed antitumor and proinflammatory macrophages ex-vivo with HDAC6 inhibitors (HDAC6i). We administered the reprogrammed macrophages intratumorally as an adoptive cell therapy (ACT) in the syngeneic SM1 murine melanoma model and patient-derived xenograft bearing NSG-SGM3 humanized mouse models. We phenotyped the tumor-infiltrated immune cells by flow cytometry and histological analysis of tumor sections for macrophage markers. We performed bulk RNA-seq profiling of murine bone marrow-derived macrophages treated with vehicle or HDAC6i and single-cell RNA-seq profiling of SM1 tumor-infiltrated immune cells to determine the effect of intratumor macrophage ACT on the tumor microenvironment (TME). We further analyzed the single-cell data to identify key cell-cell interactions and trajectory analysis to determine the fate of tumor-associated macrophages post-ACT. RESULTS: Macrophage ACT resulted in diminished tumor growth in both mouse models. We also demonstrated that HDAC6 inhibition in macrophages suppressed the polarization toward tumor-promoting phenotype by attenuating STAT3-mediated M2 reprogramming. Two weeks post-transplantation, ACT macrophages were viable, and inhibition of HDAC6 rendered intratumor transplanted M1 macrophages resistant to repolarization towards protumor M2 phenotype in-vivo. Further characterization of tumors by flow cytometry, single-cell transcriptomics, and single-cell secretome analyses revealed a significant enrichment of antitumor M1-like macrophages, resulting in increased M1/M2 ratio and infiltration of CD8 effector T-cells. Computational analysis of single-cell RNA-seq data for cell-cell interactions and trajectory analyses indicated activation of monocytes and T-cells in the TME. CONCLUSIONS: In summary, for the first time, we demonstrated the potential of reprogramming macrophages ex-vivo with HDAC6 inhibitors as a viable macrophage cell therapy to treat solid tumors.

Department

Anatomy and Regenerative Biology

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