Phase I Study Results of UF-Kure19, a CAR-T Product Manufactured in Less Than 1 Day, in Patients with Relapsed/Refractory Non-Hodgkin’s Lymphoma

CD19 CAR-T therapy has markedly improved outcomes for Non-Hodgkin's Lymphoma (NHL) patients, yet its availability is critically limited. Around 75-80% of eligible patients in the U.S., and many more globally, do not receive it. Major barriers include the complex, slow, and costly manufacturing processes with poor scalability. Additionally, treatment delays contribute to disease progression in many patients. High relapse rates (30-60% within a year) and side effects such as cytokine release syndrome and neurotoxicity are common. To overcome these issues, we developed UF-CAR, an ultra-fast CAR-T manufacturing platform that eliminates the need for ex vivo CAR-T expansion. This platform produces cost-effective CAR-T product directly from peripheral blood mononuclear cells (PBMCs) in under 24 hours. UF-CAR avoids T cell isolation thus reducing time, costs, and equipment needs. This process supports rapid, low-cost, and highly scalable CAR-T production with global applicability. We also anticipate an improved safety profile due to slower tumor-killing kinetics associated with the high proportion of naïve and early memory T cells.

We assessed the safety and efficacy of UF-Kure19, a second-generation 4-1BB CAR, in a single-arm, multi-center Phase 1 study (NCT05400109) involving 10 patients with CD19+ relapsed/refractory NHL. Participants had a median age of 64.5 years (range 45-83) and had undergone a median of 1.5 lines of therapy (range 1-7). Lymphoma subtypes included Mantle cell (n=2), Follicular (n=4), DLBCL (n=3), and Plasmablastic lymphoma (n=1). UF-Kure19 was manufactured in less than one day for all patients and met all predefined release criteria. The infused product’s naïve, central memory, effector memory, and late effector cell percentages were consistent with pre-apheresis levels, highlighting the non-expanded nature of the manufacturing process. Patients received a single dose of UF-Kure19 (17.5 million CAR-T cells) following cyclophosphamide (500 mg/m²) and fludarabine (30 mg/m²) conditioning. As of July 30, 2024, with a median follow-up of 6 months (range 1.6-14.4), the response rate was 80% (8 of 10), with all 8 responders achieving a complete metabolic response. The 2 non-responders initially showed stable disease but later experienced progression. Complete remission was sustained in all responding patients by the cutoff date. Nine patients are alive at a median of 6 months (range 1.6.-14.4) after treatment while one of the non-responders died of disease progression. The median duration of response and progression-free survival have yet to be determined, with updated patient outcomes to be presented.

CAR+ T cells expansion was detected 2 days post-infusion in all but one non-responder. Delayed kinetics of CAR+ T cell expansion was observed as compared to standard CAR-T products as the peak UF-Kure19 CAR+ T cell levels reached 7.07% (range: 2.3%-13.2%) with a peak time of 21 days (range: 6-30 days). Proliferating (Ki67+) CAR-T cells above our limit of quantification were observed only in patients achieving remission (median peak Ki67+CAR T-cells: 1.5%; range 1.4% to 8.8%). B cell aplasia was noted in all patients by day +30 and has persisted in all samples tested (median 90 days, range 30 days to 1 year). Circulating CAR-T cells (>1% of T cells) persisted in all responders, including one patient up to 12 months post-infusion.

Adverse effects included CRS in 3 (30%) patients (grade 1 n=1, grade 2 n=2), with a median onset of 9 days (range 7-11). One patient with grade 2 CRS received tocilizumab, with both grade 2 cases resolving to grade 1 or less within one day. One patient experienced ICANS (grade 3), which resolved within one day. The additional grade 3 or greater adverse events were hypoxia (n=1), neutropenia (n=8), leukopenia (n=6), lymphopenia (n=4), and a thromboembolic event (n=1).

Immunophenotyping showed that the first non-responder (plasmablastic lymphoma) had the lowest levels of CD4 T cells, naïve and early memory T cells, with reduced CCR4, CCR7, CD127, and CD28 expression, but higher KLRG1, CD38, and CD39 levels in both the apheresis and CAR products. The second non-responder (DLBCL) had a similar T cell profile to responders but lacked detectable proliferating Ki67+ CAR+ T cells post-infusion. Overall, UF-Kure19 demonstrated high efficacy and tolerability, with the potential to address current accessibility challenges and extend CAR-T therapy's reach worldwide.