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The First-In-Class Anti-AXL×CD3ε Pronectin™-Based Bispecific T-Cell Engager Is Active in Preclinical Models of Human
Nicoletta Polerà1, Antonia Mancuso1, Caterina Riillo1
1Department of Experimental and Clinical Medicine, Magna Græcia University, 88100 Catanzaro, Italy.
Abstract:
Sarcomas are heterogeneous malignancies with limited therapeutic options and a poor prognosis. We developed an innovative immunotherapeutic agent, a first-in-class Pronectin™-based Bispecific T-Cell Engager (pAXL×CD3ε), for the targeting of AXL, a TAM family tyrosine kinase receptor highly expressed in sarcomas. AXL expression was first analyzed by flow cytometry, qRT-PCR, and Western blot on a panel of sarcoma cell lines. The T-cell-mediated pAXL×CD3ε cytotoxicity against sarcoma cells was investigated by flow cytometry, luminescence assay, and fluorescent microscopy imaging. The activation and degranulation of T cells induced by pAXL×CD3ε were evaluated by flow cytometry. The antitumor activity induced by pAXL×CD3ε in combination with trabectedin was also investigated. In vivo activity studies of pAXL×CD3ε were performed in immunocompromised mice (NSG), engrafted with human sarcoma cells and reconstituted with human peripheral blood mononuclear cells from healthy donors. Most sarcoma cells showed high expression of AXL. pAXL×CD3ε triggered T-lymphocyte activation and induced dose-dependent T-cell-mediated cytotoxicity. The combination of pAXL×CD3ε with trabectedin increased cytotoxicity. pAXL×CD3ε inhibited the in vivo growth of human sarcoma xenografts, increasing the survival of treated mice. Our data demonstrate the antitumor efficacy of pAXL×CD3ε against sarcoma cells, providing a translational framework for the clinical development of pAXL×CD3ε in the treatment of human sarcomas, aggressive and still-incurable malignancies.
Insights
A novel bispecific T-cell engager targeting AXL (pAXL×CD3ε) shows significant antitumor activity against sarcoma cells. This immunotherapy agent activates T-cells and inhibits tumor growth in vivo, offering a promising new treatment for aggressive sarcomas.
Area of Science:
- Oncology
- Immunotherapy
- Molecular Biology
Background:
- Sarcomas are aggressive cancers with limited treatment options.
- AXL receptor tyrosine kinase is highly expressed in many sarcomas.
- Novel immunotherapeutic strategies are needed to improve sarcoma treatment outcomes.
Purpose of the Study:
- To develop and evaluate a novel Pronectin™-based Bispecific T-Cell Engager (pAXL×CD3ε) targeting AXL in sarcomas.
- To assess the in vitro and in vivo efficacy of pAXL×CD3ε as a potential sarcoma immunotherapy.
Main Methods:
- AXL expression analysis in sarcoma cell lines using flow cytometry, qRT-PCR, and Western blot.
- In vitro assessment of T-cell activation, degranulation, and cytotoxicity induced by pAXL×CD3ε.
- In vivo efficacy studies of pAXL×CD3ε in human sarcoma xenograft models in immunocompromised mice.
Main Results:
- High AXL expression was observed in most sarcoma cell lines.
- pAXL×CD3ε demonstrated dose-dependent T-cell activation and potent T-cell-mediated cytotoxicity against sarcoma cells.
- Combination therapy with trabectedin enhanced cytotoxicity, and pAXL×CD3ε inhibited tumor growth in vivo, improving mouse survival.
Conclusions:
- pAXL×CD3ε exhibits significant antitumor efficacy against sarcomas.
- This first-in-class agent provides a promising translational framework for clinical development in treating human sarcomas.
