Figure 5.

CD38 inhibition induced CD38-cADPR-Ca²⁺signaling suppression in CAR-T cells

(A) Schematic of CD38-cADPR signaling and following activation of Ca²⁺ signaling.

(B) Intracellular cADPR and ADPR level in control or 78C-treated CAR-T cells after coculture with NALM6 cells (cADPR, n = 10, 5 biological replicates with 2 technical replicates for each donor. ADPR, n = 8, 4 biological replicates with 2 technical replicates for each donor).

(C) Median fluorescence intensity of Fluo-4 in CAR-T cells treated with DMSO, 78C, and 78C with cADPR, ADPR, or NAADP after coculture with NALM6 cells (n = 4 biological replicates).

(D) Flow cytometric analysis of CD62L and CD45RO in CAR-T cells treated with DMSO, 78C, and 78C with cADPR, ADPR, or NAADP.

(E) Frequency of CD62L⁺ CAR-T subset in each group (n = 4 biological replicates).

(F) Frequency of CD69⁺ CAR-T cells in CAR-T cells treated with DMSO, 78C, and 78C with cADPR, ADPR, or NAADP (n = 3 biological replicates).

(G) Frequency of LAG-3⁺, TIM-3⁺, and PD-1⁺ subsets in CAR-T cells treated with DMSO, 10 μM 78C, and 10 μM 78C with 30 μM cADPR, 500 μM ADPR, or 100 μM NAADP.

(H) mRNA level of glycolysis-related transcription factors in CAR-T cells treated with DMSO, 10 μM 78C, or 10 μM 78C with 5 μM cADPR (n = 3 biological replicates).

(I) Flow cytometric analysis of CD62L and CD45RO in CAR-T cells treated with DMSO or 10 μM 8-Br-cADPR.

(J) Frequency of CD62L⁺ CAR-T subset in each group (n = 3 donors).

(K) Frequency of CD25⁺ and CD69⁺ CAR-T cells in control or 8-Br-cADPR-treated CAR-T cells after coculture with NALM6 cells (n = 3 biological replicates).

(L) Frequency of LAG-3⁺, TIM-3⁺, and PD-1⁺ subsets in control or 8-Br-cADPR-treated CAR-T cells after coculture with NALM6 cells (n = 3 biological replicates).

(M) mRNA level of glycolysis-related transcription factors in CAR-T cells treated with DMSO or 8-Br-cADPR (n = 3 technical replicates).