Background: CD19-directed CAR-T cell therapy has been demonstrated to be a valuable treatment option for relapsed/refractory B-cell non-Hodgkin’s lymphoma (r/r B-NHL). It has been shown that 39% to 97% of clinical samples of B-NHL also express BCMA. To further improve safety and efficacy, we have developed a CD19 and BCMA dual-targeting CAR-T, GC012F, manufactured on the novel next-day FasT CAR-T process, for the treatment of r/r B-NHL. The initial 3-patient data of an investigator-initiated trial (ChiCTR2100047061) was reported at EHA 2022 (#2938). Here, we present updated results of this ongoing IIT trial. Methods: From October 2021 to January 2023, nine eligible r/r DLBCL pts were enrolled and treated with a single infusion of GC012F in escalating dosing cohorts after a standard lymphodepletion regimen. The primary objectives of this study were safety and tolerability; the secondary were pharmacokinetics and efficacy. Cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS) were graded by ASBMT 2019, adverse effects were evaluated according to CTCAE 5.0. Efficacy assessment of GC012F was referred to the Lugano criteria. Expansion of CAR-T cells and CAR copy numbers were measured by flow cytometry and qPCR, respectively. Results: As of data cut-off of January 16^th, 2023, with a median of 207 days (range 94-460) of follow-up, all nine pts received single GC012F infusions at dose levels from 3.7×10⁴ to 3×10⁵ CAR-T/kg, and completed at least 3 months of follow-up. All pt lymphoma samples expressed CD19, and 6 out of 7 expressed BCMA. The median SPD was 2690.81 mm² (408.3-13325.96). Pts received a median of 2 prior lines (range 2-3) of therapy including rituximab and anthracyclines. Two pts received prior auto-HSCT. The ORR in nine pts was 100% (9/9) at month 3, CR rate was 77.8% (7/9) at month 3 and 71.4% (5/7) at month 6, respectively. To date, the longest duration of remission was 14.4 months. No dose-limiting toxicities were observed. One pt had grade 3 CRS over 2 days. No ICANS were observed. Of the 9 treated pts, ≥ grade 3 TEAEs were neutropenia (7/9), leukopenia (5/9) and thrombocytopenia (3/9). All TEAEs were resolved after treatment with standard of care and supportive care. The median peak copy number of CAR-T cells in peripheral blood was 71,000 copies/μg DNA (range 9263 -185,039), and the median peak time was 14 days (range 9 - 21). CAR-T cells were also detected in tumor biopsies from all five pts tested. Conclusions: This first-in-human trial of GC012F for the treatment of r/r B-NHL showed a manageable safety profile and promising clinical responses. The ORR was 100% at M3 with 77.8% (7/9) achieving CR. GC012F CAR-T cells were detectable in the tumor biopsies, indicating the infiltration of CAR-T cells into the tumor lesions. Study with larger cohort and longer follow-up is ongoing. Clinical trial information: ChiCTR2100047061.