CART-EGFR-IL13Ra2 in EGFR Amplified Recurrent GBM

Phase 1, Open-label Study Evaluating the Safety and Feasibility of CART-EGFR-IL13Ra2 Cells in Patients With EGFR-Amplified Recurrent Glioblastoma

This is an open-label phase 1 study to assess the safety and feasibility of autologous T cells co-expressing two CARs targeting the cryptic EGFR epitope 806 and IL13Ra2 (referred to as "CART-EGFR-IL13Ra2 cells") in patients with EGFR-amplified glioblastoma, IDH-wildtype that has recurred following prior radiotherapy.

This is a Phase 1 study evaluating the safety and feasibility of CART-EGFR-IL13Ra2 cells in a 3+3 dose escalation design as described below. • Cohort 1 (N = 3-6): will receive a single fixed dose of 1x107 CART-EGFR-IL13Ra2 cells via intrathecal administration on Day 0. This dose level will be evaluated as follows: If 1 DLT/3 subjects occurs, the study will enroll an additional 3 subjects at this dose level. If 0 DLT/3 subjects or 1 DLT/6 subjects occur, the study will advance to Cohort 2. In the event that 2 or more DLTs occur in Cohort 1, then enrollment into Cohort 1 will be stopped and the dose will be de-escalated to 5x106 CART-EGFR-IL13Ra2 cells. This de-escalated cohort will be identified as Cohort -1. • Cohort -1 (N = 3-6): will receive a single fixed dose of 5x106 CART-EGFR-IL13Ra2 cells via intrathecal administration on Day 0. This dose level will be evaluated as follows: If 0 DLT/3 or 1 DLT/3 subjects occurs, the study will enroll an additional 3 subjects at this dose level. If ≥ 2 DLTs occur at any time, enrollment onto this cohort will be stopped. In the event of 0 DLT/3 subjects or 1 DLT/6 subjects in Cohort 1, then the study will advance to Cohort 2. Cohort 2 (N = 3-6): will receive a single fixed dose of 2.5x107 CART-EGFR-IL13Ra2 cells via intrathecal administration on Day 0. This dose level will be evaluated as follows: If 1 DLT/3 subjects occurs, the study will enroll an additional 3 subjects at this dose level. If 0 DLT/3 subjects or 1 DLT/6 subjects occur, the study will advance to Cohort 3. If 2 DLTs occur at any time, enrollment in this Cohort will be stopped. If less than 6 subjects were treated at the previous dose level (Cohort 1), additional subjects will be enrolled in that Cohort to reach a minimum of 6 evaluable subjects. Cohort 3 (N = 3-6): will receive a single fixed dose of 1x108 CART-EGFR-IL13Ra2 cells via intrathecal administration on Day 0. This dose level will be evaluated as follows: If 1 DLT/3 subjects occurs, the study will enroll an additional 3 subjects at this dose level. If 0 DLT/3 subjects occurs, the study will enroll an additional 3 subjects at this dose level. If 2 DLTs occur at any time, enrollment in this Cohort will be stopped. If less than 6 subjects were treated at the previous dose level (Cohort 2), additional subjects will be enrolled in that Cohort to reach a minimum of 6 evaluable subjects. The maximum tolerated dose (MTD) is defined as the highest dose at which 0 or 1 DLT occurs in 6 evaluable subjects. The DLT observation period is 28 days post-initial treatment with CART-EGFR-IL13Ra2 cells (Day 0). In order to allow for appropriate monitoring/assessment of toxicities, the CART-EGFR-IL13Ra2 injections in the 1st and 2nd subjects in each cohort must be staggered by at least 28 days. If there are no emergent safety concerns identified in the first subject treated, subsequent subject injections within that same cohort do not need to be staggered and may occur sequentially (e.g. CART-EGFR-IL13Ra2 injections in the 2nd and 3rd subjects may occur in parallel without additional staggering requirements). Formal DLT evaluations will be performed after the 3rd subject in each cohort reaches the Day 28 safety follow-up visit, and will allow for a formal decision regarding cohort progression, expansion, or dose de-escalation. Formal DLT evaluations will be determined by the Clinical PI and Sponsor Medical Director in accordance with the definition in the protocol, Section 8.1.7. Subjects must receive the target dose of CART-EGFR-IL13Ra2 cells as per their cohort assignment in order to be considered evaluable for dose escalation decisions and MTD determination. Subjects who do not receive the dose of CART-EGFR-IL13Ra2 cells as per their cohort assignment will not be considered evaluable for this purpose and will be replaced in this cohort. However, these subjects will still be included in the overall safety analysis, as well as the analyses of secondary and exploratory objectives.

Cohort 1 (N = 3-6): will receive a single fixed dose of 1x107 CART-EGFR-IL13Ra2 cells via intrathecal administration on Day 0.

Cohort -1 (N = 3-6): will receive a single fixed dose of 5x106 CART-EGFR-IL13Ra2 cells via intrathecal administration on Day 0.

Cohort 2 (N = 3-6): will receive a single fixed dose of 2.5x107 CART-EGFR-IL13Ra2 cells via intrathecal administration on Day 0.

Cohort 3 (N = 3-6): will receive a single fixed dose of 1x108 CART-EGFR-IL13Ra2 cells via intrathecal administration on Day 0.

autologous T cells transduced with a bicistronic lentiviral vector containing a murine scFv targeting EGFR epitope 806 and a humanized scFv targeting IL13Ra2

Number of Subjects with treatment related adverse events using NCI Common Terminology Criteria for Adverse Events (CTCAE) V5.0

Inclusion Criteria: 1. Signed, written informed consent Male or female age ≥ 18 years Patients with glioblastoma, IDH-wildtype (as defined by WHO 2021 Classification of CNS Tumors) that has recurred following prior radiotherapy11. For patients with tumors harboring methylation of the MGMT promoter, at least 12 weeks must have elapsed since completion of first-line radiotherapy. Tumor tissue positive for wild-type EGFR amplification by NeoGenomics Laboratories. Archival tumor from patient's initial surgery at time of original diagnosis or recently collected tumor from time of recurrence are acceptable. Surgical tumor resection for disease control/management or tumor biopsy to confirm tumor recurrence is clinically indicated in the opinion of the physician-investigator. Adequate organ function defined as: Serum creatinine ≤ 1.5 x ULN or estimated creatinine clearance ≥ 30 ml/min and not on dialysis. ALT/AST ≤ 3 x upper limit of normal range and total bilirubin ≤ 2.0 mg/dl, except for patients in whom hyperbilirubinemia is attributed to Gilbert's syndrome (≤ 3.0 mg/dl). Left Ventricular Ejection Fraction (LVEF) ≥ 45% confirmed by ECHO/MUGA Must have a minimum level of pulmonary reserve defined as ≤ Grade 1 dyspnea and pulse oxygen > 92% on room air Karnofsky Performance Status ≥ 60%. Subjects of reproductive potential must agree to use acceptable birth control methods. Exclusion Criteria: 1. Active hepatitis B or hepatitis C infection. Any other active, uncontrolled infection. Class III/IV cardiovascular disability according to the New York Heart Association Classification. Tumors primarily localized to the brain stem or spinal cord. Severe, active co-morbidity in the opinion of the physician-investigator that would preclude participation in this study. Receipt of bevacizumab within 3 months prior to physician-investigator confirmation of eligibility. Active autoimmune disease requiring systemic immunosuppressive treatment equivalent to ≥ 10 mg daily of prednisone. Patients with autoimmune neurological diseases (such as MS or Parkinson's) will be excluded. Pregnant or nursing (lactating) women. History of allergy or hypersensitivity to study product excipients (human serum albumin, DMSO, and Dextran 40).