ABSTRACT
Leukemia encompasses a heterogeneous group of blood cancers characterized by uncontrolled proliferation and accumulation of abnormal white blood cells. The aberrant behaviors of leukemic cells enabling unchecked growth, therapeutic resistance, and metastasis arise from underlying abnormalities in their fundamental cellular biophysical properties. This comprehensive study employs a multitude of biophysical profiling techniques to investigate the morphological, mechanical, electrochemical, thermodynamic, and molecular biophysical characteristics of primary human acute myeloid leukemia (AML) cells in comparison with normal healthy hematopoietic cells. High-resolution scanning electron microscopy and super-resolution fluorescence microscopy reveal striking morphological differences and cytoskeletal disorganization within AML cells. Microfluidic deformability assays demonstrate substantially decreased ability of leukemic cells to squeeze through narrow constrictions, indicating rigidification. Patch clamp electrophysiology measures hyperpolarized plasma membrane potentials along with altered ion channel activities in AML cells versus normal counterparts. Isothermal titration and microscale thermophoresis calorimetry uncover heightened metabolic activity and mitochondrial dysfunction in leukemic viii cells. Atomic force microscopy detects nanoscale changes in the stiffness, adhesion, and topographical landscape of the leukemic cell membrane. Singlemolecule tracking methods quantify increased molecular mobility within the cytoplasm of live AML cells. Molecular dynamics simulations predict reduced structural stability of cytoskeletal proteins implicated in driving AML pathogenesis. This body of work substantially expands understanding of the diverse cellular biophysical aberrations underlying the molecular pathophysiology of AML. The quantitative biophysical profiling approaches shed light on fundamental factors enabling the uncontrolled proliferative, pro-survival, and metastatic capabilities of leukemic cells. The identified biophysical differences and signatures hold promise as novel biomarkers for improved leukemia detection, monitoring, prognosis, and personalized therapy. More broadly, this research highlights the wealth of insights into cancer cell biology and behavior gained from probing cancer cells through the multifaceted lens of biophysics. Keywords : Hematopoietic stem cells, Atomic force microscopy, Cell mechanics, Cell morphology, acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL), chronic myelogenous leukemia (CML)
