Project Funding Details

Although, great improvements have been made in the field of oncology, patients with recurrent and refractory (R/R) malignancies still maintain an unfavorable clinical outcome. Several immunotherapeutic approaches have been suggested including T and NK cells expressing a chimeric antigen receptor (CAR) but also in this setting has been proved that the immunosuppressive tumor microenvironment (TME) limits their efficacy. Furthermore, the CAR targeted antigens are mostly infrequent, shredded and often confined to a particular tumor entity. Generate an innovative and safe immunotherapeutical platform based on third-party genetically-modified (gm) NK capable to: 1. overcome the problem in the use of patient-derived cells and exploit intrinsic NK cell alloreactivity; 2. recognize "common" tumor receptors such as immune checkpoint (IC)-ligands expressed by many different tumors; 3. convert the negative signals of TME into an activating signal inducing strong antitumor response. Develop a ready-to-use donor-derived NK products armed with a broadly-reactive anti-tumor activating IC able to overcoming, rather than suffering, the inhibitory effect of TME. The IC-ligand targeting strategy offers the flexibility to treat a plethora of R/R neoplasms without re-editing the cell product for each patient. As model, neuroblastoma and lung cancer will be used. However, since IC-ligands are commonly expressed and/or strongly upregulated by most tumors, this approach can be applied to a broad number of other tumors. The project will aim to: WP1 Generate gmNK cells encoding for chimeric proteins containing an extracellular domain of IC (exIC) and an intracellular activating domain (NKG2D.4-1BB) together with an anti-NKG2A and IL15. WP2 Expand donor-derived NK cells in presence of different metabolites, capable to imprint a long-lasting metabolic reprogramming and protect them from metabolic exhaustion within TME. WP3 Characterize (phenotype and function) exIC-NK cells using static and dynamic 2D and 3D in vitro systems and mouse models. WP4 Perform a deeper analysis of TME to classify different tumor types and identify intratumoral differences between primary tumor and metastasis based on IC-ligand expression. WP5 Apply integrative data analysis together with haplotype-matching to establish the best patient/exIC-NK cell match. We expect to define a new immunotherapy capable to improve outcome of patients with R/R tumors. The success of this project will lead in the manufacturing of a ready-to-use, off-the-shelf NK product able to counteract immunosuppressive TME. A deeper analysis of TME will allow identifying intra-tumoral differences and selecting the best "personalized" NK-cellular product for each tumor and patient. This approach provides new knowledges and paving the way for a new generation of immunotherapy products circumventing several challenges and obstacles achieving long-term NK efficacy and persistence, better overall survival, guarantee economical sustainability and improve the quality-of-life of patients. This project is very innovative and with high translational impact since it uses an optimized immunotherapy able to counteract simultaneously different tumor-driven suppressive mechanisms. Our study could greatly improve cell-based immunotherapies because its mechanisms of action may allow wider applications to many patients and platforms. Furthermore, the use of third-party NK cells may represent an "off-the-shelf" product that is cost-reducing and may be standardized and applied rapidly to cancer patients.