Identifying and reprogramming softness-driven cancer stem-like cells overcomes CAR-T cell resistance in solid tumours.

Cancer stem cells are hyper-responsive sensors of the tumour microenvironment and regulate metastasis dynamics through YAP/TAZ

It is demonstrated that cancer stem cells are hyper-responsive to microenvironmental cues, which they sense and adapt to through YAP/TAZ signalling, thereby playing an essential role in breast cancer lung metastasis.

Integrated Engineering of CAR‐T Cells for Solid Tumours

This review examines the transition from single‐axis engineering to an integrated framework that addresses hurdles in sequence, and delineates how next‐generation CAR‐T cells are designed for precise spatiotemporal activation through logic‐gated and pharmacologically regulatable receptors, while being reinforced by metabolic and epigenetic reprogramming to resist TME‐driven exhaustion.

Single-Cell Profiling of Apoptotic Resistance in Patient-Derived Xenografts and Primary Circulating Tumor Cells Using TetherChip.

Tumor-Mimetic Microenvironment Primes Cancer Stemness and Drug Resistance through Nucleoskeleton-Mediated Chromatin Remodeling.

The tumor microenvironment (TME) critically regulates cancer progression by providing biochemical and biophysical cues that shape cellular behavior. However, how defined physical microenvironments govern cancer stemness and chemoresistance through mechanotransduction remains poorly understood. Here, we systematically engineered eight tumor-mimetic microenvironments by integrating serum, oxygen, and 3D compacted culture to investigate their effects on A549 non-small cell lung cancer cells. Among all conditions, cells cultured under 3D culture (PM4C) exhibited reduced cellular stiffness, enhanced expression of cancer stemness markers (EpCAM and CD44), and significantly increased resistance to cisplatin in both in vitro and nude mouse xenograft models. Transcriptomic analysis revealed that differentially expressed genes in the PM4C group were predominantly enriched in cell adhesion, mechanotransduction, stemness, and cisplatin resistance pathways. Metabolomic profiling further revealed a substantial accumulation of anaerobic metabolites associated with the maintenance of stemness. Mechanistically, the PM4C microenvironment remodeled matrix production, cell-ECM interactions, and cytoskeletal organization while inducing epigenetic reprogramming (reduced H3K9 acetylation), collectively promoting a stem-like and chemoresistant phenotype. These findings establish a direct mechanistic link between TME and cancer cell stemness, demonstrating that TME can reprogram stemness and drug responsiveness through mechano-epigenetic regulation. This work provides a mechanobiological framework for engineering physiologically relevant tumor organoids and offers new strategies for developing TME-targeted drugs and therapies.

Epigenome-Wide CRISPR-Cas9-Based Knockout Screens on Chemoresistant Cells.

Key methodological steps for achieving high-efficiency lentiviral transduction and selection are described, enabling the successful application of EPIKOL CRISPR screens in chemoresistant TNBC models.

Engineering controllable CAR T-cell therapies: from binary safety switches to programmable immunity

Chimeric antigen receptor (CAR) T-cell therapy has revolutionised cancer gene therapy, yet its expansion into solid tumours is hindered by a critical vulnerability: the autonomous, “always-on” nature of conventional CAR constructs. This unregulated activity drives severe toxicities, including cytokine release syndrome (CRS) and on-target/off-tumour damage, while constitutive signalling in hostile tumour microenvironments (TMEs) accelerates T-cell exhaustion. Early safety strategies relied on irreversible genetic “kill switches,” which sacrifice the therapeutic cell population entirely. This review traces the conceptual evolution of CAR T-cell controllability from binary elimination towards platforms enabling graded, reversible, and spatiotemporally precise regulation. We examine the transition from calibrated signalling architectures and small-molecule-regulated split-CARs to advanced optogenetic and sonogenetic controllers, detailing the biophysics of photoreceptor pairs and their preclinical efficacy. Furthermore, we explore complementary architectures, including autonomous logic-gated receptors. Finally, we propose that the optimal next-generation CAR T product will integrate calibrated signalling, external control, and context-dependent armouring to achieve truly programmable, safe, and durable cellular immunotherapy.