Phase I/II study of anti-GD2 Chimeric Antigen Receptor-Expressing T cells in pediatric patients affected by High Risk and/or relapsed/refractory Neuroblastoma and other relapsed/refractory GD2+ tumors

Prior treatment

• Phase I/II Exclusion Criteria 1. Severe, uncontrolled active intercurrent infections 2. HIV, or active HCV and/or HBV infection 3. Concurrent or recent prior therapies, before apheresis: a. Systemic steroids (at a dose equivalent to or greater 2 mg/kg prednisone) in the 2 weeks before apheresis collection. Recent or current use of inhaled/topical/non-absorbable steroids is not exclusionary. b. Systemic chemotherapy in the 3 weeks preceding apheresis collection. c. Immunosuppressive agents in the 2 weeks preceding apheresis collection d. Radiation therapy must have been completed at least 3 weeks prior to apheresis. e. I131 MIBG therapy must have been completes at least 6 weeks prior to apheresis f. Anti-GD2 murine monoclonal antibody (ch14.18 antibody) in the 2 weeks preceding apheresis collection g. Other anti-neoplastic investigational agents currently or within 30 days prior to apheresis (i.e. start of protocol therapy); h. Exceptions: Subjects receiving steroid therapy at physiologic replacement doses only are allowed provided there has been no increase in dose for at least 2 weeks prior to starting apheresis

Lab values

• Phase I/II - Patient Exclusion criteria 1. Pregnant or lactating women 2. Severe, uncontrolled active intercurrent infections 3. Active hepatitis B or hepatitis C infection 4. HIV infection 5. Rapidly progressive disease with life-expectancy < 6 weeks 6. History of grade 3 or 4 hypersensitivity to murine protein-containing products 7. Hepatic function: Inadequate liver function defined as total bilirubin > 4x upper limit of normal (ULN) or transaminase (ALT and AST) > 6 x ULN based on age and laboratory specific normal ranges 8. Renal function: serum creatinine > 3x ULN for age. 9. Blood oxygen saturation < 90%. 10. Cardiac function: Left ventricular ejection fraction lower than 45% by ECHO. 11. Marrow function: ANC lower than 500/mm3 and/or platelets lower than 20.000 (not reached by transfusion). 12. Congestive heart failure, cardiac arrhythmia, psychiatric illness, or social situations that would limit compliance with study requirements or in the opinion of the PI would pose an unacceptable risk to the subject. 13. Untreated CNS metastasis; patients with previous CNS tumor involvement that has been treated and is stable for at least 6 weeks following completion of therapy are eligible. 14. Concurrent or recent prior therapies, before infusion: a. Systemic steroids (at a dose equivalent to or greater 2 mg/kg prednisone) in the 2 weeks before infusion. Recent or current use of inhaled/topical/non-absorbable steroids is not exclusionary. b. Systemic chemotherapy in the 2 weeks preceding infusion. c. Immunosuppressive agents less than or equal to 30 days. d. Radiation therapy must have been completed at least 3 weeks prior to enrollment. e. I131-MIBG therapy must have been completes at least 6 weeks prior to enrollment f. Anti-GD2 murine monoclonal antibody (ch14.18 antibody) in the 2 weeks preceding infusion g. Other anti-neoplastic investigational agents currently or within 30 days prior to start of protocol therapy; h. Exceptions: ii. Subjects receiving steroid therapy at physiologic replacement doses only are allowed provided there has been no increase in dose for at least 2 weeks prior to starting apheresis 15. Patient-derived GD2-CART01 production failure: vitality <80%, CD3+ cells <80%, CD3+ CAR+ cells <20%, CD3+ CAR+ antitumor activity <60% in functional coculture assay at an Effector:Targetratio 1:1, viable CAR+ cells upon AP1903 exposition >20%, RCR positivity, Vector Copy Number >10, non-sterility, endotoxin contamination (> 1 EU/ml).

• Phase I - Patient Inclusion Criteria The patient must meet the following eligibility inclusion criteria to be enrolled to receive treatment in the Phase I study. 1. Diagnosis of NBL that have been treated with frontline therapy and is judged to be incurable, based upon the following criteria: a. Relapse after first-line treatment, proved by a positive 123-I-mMIBG-scan b. Persistence/progression of disease after the initiation of the upfront treatment 2. Patients must have measurable or evaluable disease at the time of treatment enrollment, as shown by bone marrow biopsy/aspirate, US or CT/MRI scan or by 123-I-mMIBG scan. 3. Recover from the toxic effect of previous chemotherapies: grade 4 and or 3 non-hematologic toxicities must have resolved to grade ≤2; if some effects of the therapies have become chronic (i.e. treatment associated thrombocytopenia), the patient must be clinically stable, according to the opinion of the treating physicians, and meet all other eligibility criteria. 4. Age: 12 months – 18 years. 5. Voluntary informed consent is given. For subjects < 18 years old their legal guardian must give informed consent. Pediatric subjects will be included in age appropriate discussion and verbal assent will be obtained for those greater than or equal to 12 years of age, when appropriate. 6. Clinical performance status: Patients > 16 years of age: Karnofsky greater than or equal to 60%; Patients less than or equal to 16 years of age: Lansky scale greater than or equal to 60%. 7. Patients of child-bearing or child-fathering potential must be willing to practice birth control from the time of enrollment on this study and for four months after receiving the preparative regimen. 8. Females of child-bearing potential must have a negative pregnancy test because of the potentially dangerous effects on the fetus.
• Phase II - Patients Inclusion criteria The patient must meet the following eligibility inclusion criteria to be enrolled to receive treatment in the Phase II study. 1. Diagnosis of NBL that have been treated with frontline therapy and is judged to be incurable, based upon the following criteria: a. Relapse after first-line treatment, proved by a positive MIBG-scan b. Persistence/progression of disease after the initiation of the upfront treatment OR 2. Diagnosis of extremely High Risk NBL at high risk of relapse, defined by stage III/IV and Myc-N amplification, at the end of the first-line treatment according to the Standard of Care, even if NED. OR 3. Diagnosis of GD2+ tumors other than Neuroblastoma, considered incurable with conventional treatments by the treating physician (Phase II only). 4. Patients with relapsed/refractory disease must have measurable or evaluable disease at the time of treatment enrollment, as shown by bone marrow biopsy/aspirate, US or CT/MRI scan or by MIBG-scan. 5. Recover from the toxic effect of previous chemotherapies: grade 4 and or 3 non-hematologic toxicities must have resolved to grade ≤2; if some effects of the therapies have become chronic (i.e. treatment associated thrombocytopenia), the patient must be clinically stable, according to the opinion of the treating physicians, and meet all other eligibility criteria. 6. Age: 12 months –35 years. 7. Voluntary informed consent is given. For subjects < 18 years old their legal guardian must give informed consent. Pediatric subjects will be included in age appropriate discussion and verbal assent will be obtained for those greater than or equal to 12 years of age, when appropriate. 8. Clinical performance status: Patients > 16 years of age: Karnofsky greater than or equal to 60%; Patients less than or equal to 16 years of age: Lansky scale greater than or equal to 60%. 9. Patients of child-bearing or child-fathering potential must be willing to practice birth control from the time of enrollment on this study and for four months after receiving the preparative regimen. 10. Females of child-bearing potential must have a negative pregnancy test because of the potentially dangerous effects on the fetus