Development and Manufacturing Optimization of CAR NKT Cell Therapy for High-Risk Neuroblastoma

• Applicant: Dr. Tina Trautmann
• Subject Area: Pediatric and Adolescent Medicine ; Hematology, Oncology
• Term: since 2023
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 524613699

Neuroblastoma (NB) is the most common extracranial solid malignancy of childhood and accounts for 15% of cancer-related deaths in pediatrics. Half of all children with NB present with high-risk disease, of which less than 50% attain long-term remissions. Hence, new therapies are urgently needed to improve the survival of affected patients. Adoptive cell therapy (ACT) with genetically engineered T cells has excelled with unprecedented success in patients with leukemias and lymphomas. However, ACT in solid neoplasms like NB has shown limited antitumor efficiency in clinical testing. The most developed ACT in NB is GD2-targeting chimeric antigen receptor (CAR) T cell therapy. Despite the excellent tolerability without on-target/off-tumor toxicity, GD2-CAR T cells have yielded overall disappointing antitumor responses in the clinic. Major roadblocks to better efficacy are in part caused by (1) the restricted availability of tumor-specific antigens, (2) the insufficient expansion and persistence of T cells, and (3) the immunosuppressive tumor microenvironment. More refined CAR engineering and manufacturing could overcome some of these challenges and enhance the efficacy of future CAR products in solid tumors. We will utilize a 2nd generation CAR that targets a new antigen called glypican-2 (GPC2) to develop a GPC2-CAR NKT cell therapy against high-risk NB. We hypothesize that the use of cytokine combinations in the manufacturing process and constitutively expressed on the NKT cell surface can enhance the effector cell function of GPC2-CAR NKT cells and serve as an orthogonal approach to enhance antitumor potency of ACT in NB. Our primary objective is to optimize the manufacturing conditions with a combination of the cytokines IL-7, IL15, IL-18, and IL-21 to generate potent GPC2-CAR NKT cells. These cells will be subjected to thorough molecular and phenotypic characterization and functional testing in vitro and in vivo. Our secondary objective is to use the gained information and develop GPC2-CAR NKT cells with constitutive membrane-bound cytokine expression to maintain the cytokine priming effect beyond the stage of cell manufacturing. The successful completion of this project will yield a GPC2-targeted NKT CAR product for further clinical development to improve the treatment and outcomes of children with high-risk NB.

• DFG Programme: WBP Fellowship
• International Connection: USA
• Host: Dr. Rosa Nguyen, Ph.D.