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Combination immunotherapy including OncoVEXmGMCSF creates a favorable tumor immune micro-environment in transgenic BRAF murine melanoma

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Abstract

Talimogene Laherparepvec (OncoVEXmGMCSF), an oncolytic virus, immune checkpoint inhibitor anti-programmed cell death protein 1 (anti-PD1), and BRAF inhibition (BRAFi), are all clinically approved for treatment of melanoma patients and are effective through diverse mechanisms of action. Individually, these therapies also have an effect on the tumor immune microenvironment (TIME). Evaluating the combination effect of these three therapies on the TIME can help determine when combination therapy is most appropriate for further study. In this study, we use a transgenic murine melanoma model (Tyr::CreER; BRAFCA/+; PTENflox/flox), to evaluate the TIME in response to combinations of BRAFi, anti-PD1, and OncoVEXmGMCSF. We find that mice treated with the triple combination BRAFi + anti-PD1 + OncoVEXmGMCSF have decreased tumor growth compared to BRAFi alone and prolonged survival compared to control. Flow cytometry shows an increase in percent CD8 + /CD3 + cytotoxic T Lymphocytes (CTLs) and a decrease in percent FOXP3 + /CD4 + T regulatory cells (Tregs) in tumors treated with OncoVEXmGMCSF compared to mice not treated with OncoVEXmGMCSF. Immunogenomic analysis at 30d post-treatment shows an increase in Th1 and interferon-related genes in mice receiving OncoVEXmGMCSF + BRAFi. In summary, treatment with combination BRAFi + anti-PD1 + OncoVEXmGMCSF is more effective than any single treatment in controlling tumor growth, and groups receiving OncoVEXmGMCSF had more tumoral infiltration of CTLs and less intratumoral Tregs in the TIME. This study provides rational basis to combine targeted agents, oncolytic viral therapy, and checkpoint inhibitors in the treatment of melanoma.

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Data availability

All expression data is available in supplemental files. For any other data please email corresponding author.

Abbreviations

4-HT:

4-Hydroxytamoxifen

anti-PD1:

Anti-programmed cell death protein 1

ASBL-2:

Animal biosafety level 2

BRAFi:

BRAF inhibition

CTLs:

Cytotoxic T Lymphocytes

CA:

Conditionally active

CUIMC:

Columbia university irving medical center

FFPE:

Formalin-fixed paraffin-embedded

gC:

Glycoprotein C

GM-CSF:

Human granulocyte–macrophage colony-stimulating factor

HSV-1:

Herpes simplex virus type 1

IACUC:

Institutional animal care and use committee

IP:

Intraperitoneal injection

IT:

Intratumoral injection

PBS:

Phosphate buffered saline

RT-PCR:

Transcription-polymerase chain reaction

TIME:

Tumor immune microenvironment

Tregs:

T regulatory cells

OncoVEXmGMCSF :

Talimogene laherparepvec

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Acknowledgments

We would like to thank the Human Immune Monitoring Core (HIMC) and Molecular Pathology Core at Columbia University Irving Medical Center for their help in processing the samples. We would also like to thank the Institute of Comparative Medicine for their help in caring for the animals.

Funding

The authors of this publication were supported by the National Institutes of Health through Grant Numbers R01FD006108, Project 1R01CA260375-01 (Y.M. Saenger), and KL2TR001874 (R.D. Gartrell). F. Lozano is supported by Spanish Ministerio de Economía y Competitividad (MINECO) through grant SAF-2016–80535-R co-financed by European Development Regional Fund. I. Simoes was supported by Fundação para a Ciência e a Tecnologia through a fellowship (SFRH/ BD/75738/2011). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. Yvonne Saenger is also supported by an Irving Assistant Professorship at Columbia University’s NIH/NCATS CTSA Program hub: UL1TR001873 and the Amgen-CUMC-MRA (Melanoma Research Alliance) Established Investigator Academic-Industry Partnership Award. Robyn Gartrell is also supported by Swim Across America. The funding sources had no role preparation of the manuscript or the decision to submit for publication.

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Author notes

  1. Robyn D. Gartrell and Zoë Blake are equally contributed for this work.

Authors and Affiliations

  1. Department of Pediatrics, Columbia University Irving Medical Center, 1130 St. Nicholas Avenue, ICRC 916A, New York, NY, 10032, USA

    Robyn D. Gartrell & Hanna E. Minns

  2. Department of Medicine, Columbia University Irving Medical Center, 630 W 168th Street, PS 9-428, New York, NY, 10032, USA

    Zoë Blake, Emanuelle M. Rizk, Manas Mondal, Samuel W. Wang, Benjamin T. Fullerton & Yvonne M. Saenger

  3. Department of Dermatology, Columbia University Irving Medical Center, 1150 St. Nicholas Avenue, Russ Berrie Medical Science Pavillion Room 307, New York, NY, 10032, USA

    Rolando Perez-Lorenzo

  4. Department of Pathology and Cell Biology, Columbia University Irving Medical Center, 630 W 168th Street, New York, NY, 10032, USA

    Stuart P. Weisberg

  5. Immunoreceptors del Sistema Innat I Adaptatiu, Institut d’Investigacions Biomediques August Pi I Sunyer, Barcelona, Catalunya, Spain

    Ines Simoes & Francisco Lozano

  6. Herbert Irving Comprehensicve Cancer Center, Columbia University Irving Medical Center, 161 Fort Washington Avenue, New York, NY, 10032, USA

    Camden Esancy

  7. Department of Medicine, Mount Sinai Hospital, 1468 Madison Avenue, New York, NY, 10029, USA

    Yichun Fu

  8. University of North Carolina School of Medicine, 140 W Franklin Street, Unit 506, Chapel Hill, NC, 27516, USA

    Danielle R. Davari

  9. Valegos College of Physicians and Surgeons, Columbia University, 630 W 168th Street, New York, NY, 10032, USA

    Luke Barker & Grace Finkel

  10. Department of Biostatistics, Columbia University Irving Medical Center, 722 W 168th Street, Room 646, New York, NY, 10032, USA

    Codruta Chiuzan

  11. Department of Pathology, University of British Columbia, Vancouver, BC, Canada

    Basil Horst

  12. Servei d’Immunologia, Hospital Clínic de Barcelona, Barcelona, Spain

    Francisco Lozano

  13. Departament de Biomedicina, Universitat de Barcelona, Barcelona, Spain

    Francisco Lozano

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  1. Robyn D. Gartrell
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Contributions

RG, ZB, and YS contributed to study design and conception and developed the first draft of the manuscript. ER, RPL, SW, MM, HM, SW, and BF contributed to material preparation and creation of figures and tables. Data collection and analysis were performed by RG, ZB, FL, CC, and YS. RG, ZB, IS, CE, YF, DD, LB, GF, HM, ER, SW, BF, BH, and YS discussed the manuscript sections and contributed with updated references. All authors commented on previous versions of the manuscript and all authors read and approved the final manuscript.

Corresponding author

Correspondence to Yvonne M. Saenger.

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Authors have no disclosures.

Ethical Approval

All animal experiments were performed in accordance with institutional and national guidelines and were approved by the Institutional Animal Care and Use Committee (IACUC) at Columbia University Irving Medical Center (CUIMC).

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Gartrell, R.D., Blake, Z., Rizk, E.M. et al. Combination immunotherapy including OncoVEXmGMCSF creates a favorable tumor immune micro-environment in transgenic BRAF murine melanoma. Cancer Immunol Immunother 71, 1837–1849 (2022). https://doi.org/10.1007/s00262-021-03088-y

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