Single-cell Transcriptome Analysis Reveals Mechanical Properties of Leukemic Stem Cells in AML.

Leukemic stem cells (LSCs) are crucial drivers of leukemia, yet the mechanisms underlying their function remain poorly understood. Here, we investigated the physical properties of LSCs and their potential role in LSCs function. single-cell RNA-sequencing analysis has revealed that small-sized LSCs aberrantly overexpress ALDH1A1, a crucial regulator of stem cell function. Lastly, ALDH1A1 inhibition increases cell stiffness and suppresses leukaemogenesis in patient-derived xenograft models. Our findings establish a novel link between the biomechanics of LSCs and their function in leukemia, providing a compelling rationale for targeting therapy-resistant LSCs by inhibiting G3BP. Overall, our study provides valuable insights into the mechanisms of LSCs function and has significant implications for the development of more effective therapies for this devastating disease. Overall design: In order to explore the mechanism of the function in mechanical properties of LSCs, we isolated small cells and large cells derived from bone marrow lin- cells of 4 AML patients.For the scRNA-seq analysis of human lin- small and large cells ,we pooled 4 sample from the small group in one tube and 4 sample from the large group in another tube. We then counted and resuspended the pooled cells at a concentration of 1000 cells/µL, targeting an estimated 6000 cells per library, following the instructions of the Single Cell 3' Solution v3 Kit (10X Genomics). Single-cell libraries were constructed strictly according to the manufacturer's standard protocol. Libraries were sequenced on the Illumina NovaSeq 6000 System.