NEO-TIL Developing durable novel TIL therapies for lung cancer
Abhishek Tomar (1), Rui Li (1), Alan Soo (2), Anne Marie Quinn (3), Paul Loftus (4), Michael Kerin (1), Laura R Barkley (1)
(1) Discipline of Surgery, School of Medicine, Lambe Institute for Translational Research, University of Galway, (2) Department of Cardiothoracic Surgery, Cardiothoracic Surgeon, University Hospital Galway, (3) Consultant Histopathologist, Head of Department of Anatomic Pathology, University Hospital Galway, (4) Orbsen Therapeutics Ltd., Galway Business Park, Galway
Introduction
Lung cancer is among the leading causes of cancer mortality worldwide. The advancement in therapies using monoclonal antibody-based immune checkpoint blockade (ICB) has shown minor improvements in overall survival. Regardless of these improvements, the majority of patients suffer cancer progression either after first-line treatment only or combined treatment with ICB. The development of novel personalized treatment is an urgent need for prolonged tumour regression to enhance the survival rates of lung cancer patients. Adoptive Cell Transfer of polyclonal tumour infiltrating lymphocytes (ACT-TIL) is a potential treatment option for lung cancer patients, due to lung cancers' high mutational load. This study aims to develop a process of successful TIL isolation and expansion from lung cancer tissue and generate a biobank of patient-specific tumour-reactive TILs.
Materials and methods
Ethical approval is in place to obtain cancer tissue from lung cancer patients undergoing surgery at University Hospital Galway. Lung tumour tissue is sliced into small fragments of 1-2 mm2 and seeded into TIL media supplemented with interleukin-2 (IL-2). The TILs are grown for 15 days, cell counts are performed. After expansion, TILs are characterized by flow cytometry analysis to measure T-cell markers (CD3, CD4, and CD8) and T-cell exhaustion markers (PD-1, TIM-3, and LAG-3). Autologous tumour cell lines (TCLs) are established from the same piece of lung cancer tissue. Co-culture experiments with TILs and autologous TCLs are performed and IFN-ϒ release is measured (ELISA) to assess TIL tumour reactivity.
Results and discussion
Using this process, we have isolated TILs from 28 lung cancer patients. We have determined the growth kinetics of these TIL populations. These cells are positive for T cell markers CD3, CD4, and CD8. They also express inhibitory receptors PD-1, LAG-3, TIM-3, which are typically expressed on chronically stimulated CD8+ T cells and would suggest these TILs are tumour reactive. Indeed, initial co-culture experiments show an increase in IFN-ϒ release when TILs are co-cultured with autologous tumour cells, indicating TILs are tumour reactive.
Conclusion
We have developed a research-based process that enables the isolation and growth of TILs from lung cancer tissue. Future studies will advance this TIL expansion process to a GMP-compliant process with the aim of developing a precision medicine for Irish lung cancer patients in the West of Ireland.