Phase 1 study of chidamide in combination with venetoclax, azacitidine, aclarubicin, cytarabine and G-CSF for refractory/relapsed acute myeloid leukemia: Clinical safety, efficacy and correlative analysis

For scRNA-seq analysis, bone marrow mononuclear cells (BMMCs) from four consecutively enrolled patients (July–September 2023) were processed using the 10x Genomics platform. Patients selected for scRNA-seq analysis were consecutively enrolled during the study period to minimize selection bias. Their diverse clinical outcomes provide a valuable spectrum for correlating transcriptomic changes with treatment efficacy. Overall design: BM cells from patients with AML were collected in EDTA tubes and diluted 2:1 in ice-cold FACS buffer. Mononuclear cell separation was accomplished using density centrifugation media (Ficoll-Paque; GE Healthcare Life Sciences) at a 1:1 ratio with BM cells. Subsequently, the cell suspension, with a concentration ranging from 700 to 1,200 cells/µl and a cell viability exceeding 85%, was loaded onto the Chromium Single Cell Controller (10× Genomics) to generate single-cell gel beads in the emulsion, following the manufacturer's protocol and employing the Chromium Single Cell 3' Library and Gel Bead Kit V3.1 (10× Genomics, PN1000268). The libraries were sequenced on an Illumina NovaSeq 6,000 sequencer.