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814 Enhancing anti-tumor response in ICI-refractory non-small cell lung cancer through intravenous administration of oncolytic adenovirus armed with hTNFα and hIL-2 in combination with aPD-1 blockade
  1. Tatiana V Kudling1,
  2. James Clubb1,2,
  3. Santeri A Pakola1,
  4. Dafne Alves Quixabeira1,2,
  5. Iris Lähdeniemi3,4,
  6. Camilla Heiniö1,
  7. Victor Arias1,
  8. Susanna AM Grönberg-Vähä-Koskela1,5,
  9. Riikka Havunen1,2,
  10. Victor Cervera-Carrascon1,2,
  11. Joao Manuel Santos1,2,
  12. Eva Sutinen4,6,7,
  13. Jari Räsänen8,
  14. Kristian Borenius4,8,
  15. Mikko Mäyränpää9,
  16. Eero Altonen10,
  17. Suvi Sorsa1,2,
  18. Otto Hemminki1,
  19. Anna Kanerva1,
  20. Emmy W Verschuren3,4,
  21. Ilkka Ilonen4,8 and
  22. Akseli Hemminki1,2,5
  1. 1Cancer Gene Therapy Group, Translational Immunology Research Program, University of Helsinki, Helsinki, Finland
  2. 2TILT Biotherapeutics Ltd., Helsinki, Finland
  3. 3Translational Lung Cancer Research Group, Institute for Molecular Medicine Finland (FIMM), HiLIFE, University of Helsinki, Helsinki, Finland
  4. 4iCAN Digital Precision Cancer Medicine Flagship, Helsinki, Finland
  5. 5Comprehensive Cancer Center, Helsinki University Hospital, Helsinki, Finland
  6. 6Individualized Drug Therapy Research Program, Faculty of Medicine, University of Helsinki, Helsinki, Finland
  7. 7Department of Pulmonary Medicine, Heart and Lung Center, Helsinki University Hospital, Helsinki, Finland
  8. 8General Thoracic and Esophageal Surgery, Heart and Lung Center, Helsinki University Hospital and Faculty of Medicine, University of Helsinki, Helsinki, Finland
  9. 9Pathology, University of Helsinki and Helsinki University Hospital (HUSLAB), Helsinki, Finland
  10. 10Institute for Molecular Medicine Finland (FIMM), HiLIFE, University of Helsinki, Helsinki, Finland
  • Journal for ImmunoTherapy of Cancer (JITC) preprint. The copyright holder for this preprint are the authors/funders, who have granted JITC permission to display the preprint. All rights reserved. No reuse allowed without permission.


Background Despite significant achievements in targeted and immunotherapies, including immune checkpoint inhibitors (ICI), treatment outcomes for advanced non-small cell lung cancer (NSCLC) remain challenging, particularly in patients who develop therapy resistance. This study explores the efficacy of a combination therapy involving aPD-1 checkpoint blockade and Ad5/3-E2F-d24-hTNFα-IRES-hIL-2 (a.k.a. TILT-123), a serotype chimeric oncolytic adenovirus armed with tumor necrosis factor alpha (hTNFα) and interleukin 2 (hIL-2), administered intravenously. The aim of this study is to investigate the potential of this approach for overcoming checkpoint resistance and improving anti-tumor response.

Methods We utilized immunocompetent murine pre-clinical models, including ICI-refractory subcutaneous model LLC1 and orthotopic metastasis-prone histopathology-specific NSCLC adenosquamous carcinoma model (ASC) as well as clinical samples obtained from the patients undergoing surgical resection for NSCLC. We evaluated tumor growth, cells activation, cytokine profiles, and immune cell populations within the tumor microenvironment. Additionally, a depletion study was conducted to assess the role of key immune cells in therapeutic efficacy.

Results The combination treatment of aPD-1 checkpoint blockade with intravenous administration of armed oncolytic adenovirus demonstrated a significant reduction in tumor growth, even in the presence of neutralizing antibodies against the viral treatment. This therapy led to an increased activation of cytotoxic tumor-infiltrating lymphocytes, including tumor-specific CD8+ T cells. Furthermore, the treatment resulted in a decrease in immunosuppressive tumor-associated macrophages, enhanced dendritic cell maturation, and expansion of the tumor-specific memory T cell compartment.

Conclusions Our findings highlight the potential of intravenously delivered oncolytic adenovirus armed with hTNFα and hIL-2 in combination with aPD-1 checkpoint blockade to overcome checkpoint resistance and improve treatment outcomes in advanced NSCLC. This approach is promising for addressing the major unmet clinical need in patients with checkpoint refractory/resistant NSCLC and supports further investigation in clinical trials.

Ethics Approval All animal experiments were approved by the Provincial Government of Southern Finland and the Experimental Animal Committee of the University of Helsinki (license number ESAVI/12559/2021). Cancer samples were collected from patients undergoing surgical resection at Helsinki University Central Hospital (HUS, Helsinki, Finland). Sample collection was approved by the HUS Ethics Committee (47§/17.3.2021; HUS/552/2021), and study permits were obtained (17.05.2021; reference number HUS/259/2021). Written informed consent was obtained from all participants.

This is an open access article distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited, appropriate credit is given, any changes made indicated, and the use is non-commercial. See

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