Yanteng Xu

Hepatocyte-like cells (HLCs) represent a promising therapeutic approach for acute liver failure (ALF), offering an alternative to primary hepatocytes, which are in short supply. Given the viscoelastic tissue properties of the liver and the growing recognition of the mechanical cues in physiology and disease, we investigated how ALF-induced changes in the liver's mechanical microenvironment might affect HLC-based cell therapy. Here, driven by specific disease-derived mechanical cues, we designed bioinspired hydrogel microspheres with tunable liver viscoelasticity to mimic the mechanical niche of healthy versus ALF liver tissue. We show that a fast-relaxation hydrogel niche mimicking the healthy liver guides the efficient differentiation of human adipose-derived mesenchymal stem cells (hADSCs) into functional HLCs by suppressing the formation of abnormal stress fibers and pathological YAP nuclear translocation through the ROCK axis. In both the carbon tetrachloride-induced and partial hepatectomy-induced ALF mouse models, these mechanically preconditioned HLCs exhibit significant therapeutic efficacy, promoting robust liver regeneration and acting as active mechanical regulators to suppress aberrant host mechanotransduction and remodel the diseased microenvironment. This disease mechanobiology-oriented strategy provides insights into the design of viscoelastic biomaterials for regenerative medicine and demonstrates a promising approach for enhancing cell therapy outcomes in liver failure.

Postoperative immune suppression, driven by transforming growth factor β (TGFβ)-mediated dendritic cell (DC) dysfunction, significantly impairs antigen-specific T cell responses and elevates cancer recurrence risk. A key mechanism is TGFβ-mediated lysosomal incompetence in DCs, resulting from loss of lysosomal prosaposin (pSAP). Herein, we developed an in vivo-generated DC vaccine, BAIT (boosting antigen-delivering immunovaccine for T cell priming), which integrated DC recruitment, antigen delivery, and immune activation within a single platform. Inspired by arginine metabolism-driven lysosomal biogenesis and acidification, BAITs were designed with an arginine-enriched self-assembling peptide backbone, in which tumor lysate antigens and CCL20 co-assembled via coordination interactions. When administered early post-surgery, BAITs recruited DCs, promoted antigen internalization, and activated the mTOR pathway to upregulate pSAP expression. This BAIT-driven DC modulation reversed TGFβ-induced impairments in antigen digestion and MHC-peptide complex formation. To optimize precision immunotherapy, we employed a functional precision medicine approach by customizing tumor lysate preparations induced via ferroptosis, necroptosis, or apoptosis. In a murine postoperative cancer model, Apo-BAITs formulated with apoptotic lysates achieved an 85.5% reduction in tumor burden. These findings demonstrate that BAITs target the lysosomal mTOR-pSAP axis to restore DC function, providing a personalized postoperative cancer vaccine strategy to counter surgery-induced immune suppression.

Castration-resistant prostate cancer (CRPC) remains lethal due to adaptive resistance mechanisms such as stress-induced autophagy and NF-κB survival signaling. Here, an injectable fiber-in-hydrogel depot is developed for sequential delivery of a multi-enzyme nanozyme (cobalt-epigallocatechin gallate coordination nanozyme, CoNZ) and the lysosomal inhibitor chloroquine (CQ). The preferentially released CoNZ catalytically generates reactive oxygen species, including hydroxyl radicals, and oxygen in situ, inflicting oxidative damage while relieving tumor hypoxia. Simultaneously, it depletes antioxidants (glutathione, NADPH) and impedes NF-κB nuclear translocation, priming CRPC cells for apoptosis. The subsequently released CQ impedes enzymatic degradation or cleavage in endolysosomes and autolysosomes and blocks TLR9/NF-κB signaling, preventing tumor cells from repairing damage or activating pro-survival pathways. In vitro and in vivo, this two-pronged approach synergistically overcomes CRPC's defenses, achieving markedly enhanced cancer cell apoptosis and ∼80% tumor suppression (with occasional complete regression), resulting in an approximately 10-fold reduction in final tumor volume compared to the untreated control, without systemic toxicity, far surpassing single or co-administered treatments. This work demonstrates a spatiotemporally orchestrated combination of nanocatalytic therapy and lysosomal inhibition that dismantles CRPC's resistance mechanisms, highlighting a broadly applicable paradigm for overcoming therapeutic resistance in aggressive cancers.

Acute liver failure (ALF) is a highly lethal condition characterized by massive tissue necrosis, excessive oxidative stress, and serious inflammatory storms, necessitating prompt medical intervention. Although hepatocyte-like cells (HLCs) derived from mesenchymal stromal/stem cells (MSCs) offer a promising alternative cell source for hepatocyte therapy, their low in-vivo integration and differentiation efficiency may compromise the eventual therapeutic efficacy. To this end, MSCs are bioengineered into multicellular spheroids in the present study. The proteomic analyses and experimental results reveal that extracellular vesicles (EVs) derived from these MSC spheroids (SpEV) contain abundant highly expressed bioactive proteins and can be efficiently endocytosed by recipient cells, resulting in enhanced pro-angiogenic and antioxidative effects. In addition, MSC spheroids exhibit superior hepatic cell differentiation compared to an equivalent number of dissociated single MSCs, particularly when being co-cultured with hexagonally patterned endothelial cells in a liver lobule-like arrangement. Following orthotopic implantation in the mouse model, the enhanced paracrine effects of SpEV, combined with an immunoregulatory decellularized extracellular matrix hydrogel carrier and functional artificial liver lobules (ALL), synergically contribute to the effective amelioration of ALF, highlighting the substantial potential for clinical translation.

Non-nucleotide stimulators of interferon gene (STING) agonists hold promise as immunotherapeutic agents for postsurgical adjuvant treatment of tumors. However, their limited effect duration hampers therapeutic effectiveness, necessitating prolonged administration of multiple doses that heightens infection risk and impacts patient compliance. Here, we develop an implantable dual-drug depot in a sandwich-like configuration, with a non-nucleotide STING agonist (MSA-2) in the outer layers of 3D-printed scaffolds and an immunogenic apoptosis inducer (doxorubicin, DOX) in the inner layer of electrospun fibers. We discover that MSA-2 can elicit endoplasmic reticulum stress-mediated and general immunogenic apoptosis of cancer cells. The stimulations with tumor-associated antigens and damage-associated molecular patterns from cancer cells, along with proinflammatory factors secreted by matured dendritic cells and M1-polarized macrophages, can depolymerize intracellular microtubules guiding activated STING trafficking towards lysosomes for degradation. Collectively, the dual-drug depots can initiate a long-lasting cascaded immunotherapy and chemotherapy, suppressing postsurgical tumor recurrence and metastasis.

Equipping multiple functionalities on adoptive effector cells is essential to overcome the complex immunological barriers in solid tumors for superior antitumor efficacy. However, current cell engineering technologies cannot endow these functionalities to cells within a single step because of the different spatial distributions of targets in one cell. Here, we present a core-shell anti-phagocytosis-blocking repolarization-resistant membrane-fusogenic liposome (ARMFUL) to achieve one-step multiplexing cell engineering for multifunctional cell construction. Through fusing with the M1 macrophage membrane, ARMFUL inserts an anti-CD47 (aCD47)–modified lipid shell onto the surface and simultaneously delivers colony-stimulating factor 1 receptor inhibitor BLZ945-loaded core into the cytoplasm. The surface-presenting aCD47 boosts macrophage's phagocytosis against the tumor by blocking CD47. The cytoplasm-located BLZ945 prompts its polarization resistance to M2 phenotype in the immunosuppressive microenvironment via inactivating the intracellular M2 polarization signaling pathway. This ARMFUL provides a versatile cell engineering platform to customize multimodal cellular functions for enhanced adoptive cell therapy.

As a currently common strategy to treat cancer, surgical resection may cause tumor recurrence and metastasis due to residual postoperative tumors. Herein, an implantable sandwich‐structured dual‐drug depot is developed to trigger a self‐intensified starvation therapy and hypoxia‐induced chemotherapy sequentially. The two outer layers are 3D‐printed using a calcium‐crosslinked mixture ink containing soy protein isolate, polyvinyl alcohol, sodium alginate, and combretastatin A4 phosphate (CA4P). The inner layer is one patch of poly (lactic‐co‐glycolic acid)‐based electrospun fibers loaded with tirapazamine (TPZ). The preferentially released CA4P destroys the preexisting blood vessels and prevents neovascularization, which obstructs the external energy supply to cancer cells but aggravates hypoxic condition. The subsequently released TPZ is bioreduced to cytotoxic benzotriazinyl under hypoxia, further damaging DNA, generating reactive oxygen species, disrupting mitochondria, and downregulating hypoxia‐inducible factor 1α, vascular endothelial growth factor, and matrix metalloproteinase 9. Together these processes induce apoptosis, block the intracellular energy supply, counteract the disadvantage of CA4P in favoring intratumor angiogenesis, and suppress tumor metastasis. The in vivo and in vitro results and the transcriptome analysis demonstrate that the postsurgical adjuvant treatment with the dual‐drug‐loaded sandwich‐like implants efficiently inhibits tumor recurrence and metastasis, showing great potential for clinical translation.