Zhi Zhu

Targeted protein degradation (TPD) has redefined the therapeutic landscape by shifting the focus from merely inhibiting protein function to actively eliminating disease‐causing proteins. However, the clinical translation of TPD agents remains hampered by their inherent physicochemical limitations, including poor solubility, membrane impermeability, and lack of tissue selectivity. Nanotechnology offers a strategic pathway to overcome these barriers. This review examines the convergence of TPD and nanotechnology, highlighting how diverse nanoplatforms—ranging from lipid and polymeric nanoparticles to inorganic carriers and biomimetic systems—can address fundamental delivery challenges. We discuss key design strategies such as physical encapsulation, chemical conjugation, and carrier‐free self‐assembly between nanocarrier‐assisted TPD—where nanoparticles improve degrader pharmacokinetics—and nanostructure‐integrated TPD—where the nanoscaffold directly participates in ternary complex formation, and explore how engineering the nano‐bio interface enables precise control over cellular uptake, intracellular trafficking, and ternary complex formation. We also critically assess current translational hurdles, including manufacturing complexity, biosafety concerns, and tumor heterogeneity, while offering perspectives on how rational design and interdisciplinary collaboration can accelerate clinical adoption. By reimagining protein degraders as components of intelligent nano‐systems, nano‐TPD holds the potential to transform undruggable targets into actionable therapeutic opportunities.

Extracellular vesicles (EVs) have emerged as promising biomarkers for monitoring physiological homeostasis and pathological progression. However, current analytic methods face limitations in preserving spatial information about EVs and their intricate connections to parental and recipient cells. Here, we present Spatial-EV-seq, a method for in situ spatial profiling of EVs within their native microenvironmental context. Spatial-EV-seq uses an antibody-engineered capture interface to preserve EVs' spatial distribution, followed by rolling circle amplification with EV surface-binding aptamers, enabling fluorescence imaging and molecular profiling of individual EVs. The method integrates ultrasensitive EV profiling, molecular subtyping and high-resolution spatial mapping with transcriptomics to resolve location-specific EV-cell communication networks. In an anti-PD1-treated breast cancer mouse model, we uncover a spatially orchestrated immunosuppressive axis: PDL1⁺ EV-enriched zones drive CD8⁺ T cell dysfunction, establishing immune-privileged niches, whereas PDL1⁺ EV-depleted regions preserve immunocompetence and therapeutic sensitivity. Spatial-EV-seq offers insights into EV-mediated mechanisms and unlocks avenues for precision diagnostics and therapeutics.

Tissues serve as the functional units of multicellular organisms with intricate spatial organizational complexity. Next-generation sequencing (NGS)-based spatially resolved transcriptomics (SRT) delineates in situ gene expression heterogeneities, establishes high-fidelity associations between transcripts and spatial pixels by segmenting a tissue section into spatial pixels, and elucidates the pivotal roles of cellular spatial organization in biological processes and complex pathological mechanisms. The synergistic integration of nucleic acid barcoding and high-throughput sequencing technologies has rapidly advanced the development of these spatial modalities regarding throughput, resolution, cost-effectiveness, and sensitivity. In this review, we summarize the state-of-the-art sequencing-based spatial transcriptomics, with a primary emphasis on the methodologies. We first introduce two major categories of typical nucleic acid barcoding platforms (in situ barcoding-based and barcoding array-based) used for spatial localization and transcriptome profiling. Then, the burgeoning trend of spatial transcriptomics towards spatiotemporal transcriptomics, which integrates SRT with the temporal dimension to provide more holistic landscapes of gene expression networks, is discussed. We also highlight spatiotemporal transcriptomics based on metabolic RNA labeling that provides unprecedented resolution to resolve transcriptome-wide dynamics in space and time. The emerging applications of these technologies in providing mechanistic insights into complex pathological mechanisms are also discussed. Finally, the perspectives on current bottlenecks and future direction of spatiotemporal transcriptomics are provided.

Targeted protein degradation (TPD) has emerged as a transformative therapeutic strategy, but most existing approaches are restricted to intracellular proteins. Lysosome‐targeting chimeras (LYTACs) expand the scope of TPD by enabling the selective degradation of extracellular and membrane‐associated proteins through the endosomal–lysosomal pathway. This review provides a comprehensive overview of LYTAC technology, focusing on its molecular architecture and mechanism of action. We systematically examine the evolution of LYTACs from two key design dimensions: (1) the diversity of protein‐of‐interest (POI) binders, including antibodies, aptamers, small molecules, peptides, and DNA scaffolds; and (2) the expanding repertoire of lysosomal targeting receptors (LTRs), from classical receptors, cation‐independent mannose‐6‐phosphate receptor (CI‐M6PR) and asialoglycoprotein receptor (ASGPR), to emerging tissue‐restricted, multifunctional, and programmable entry routes. Finally, we discuss strategies to enhance degradation efficiency, pharmacokinetics/pharmacodynamics (PK/PD) profiles, and address key challenges for clinical translation, offering a framework for the rational design of next‐generation LYTAC therapeutics.

Cellular softness acts as a mechanical immune checkpoint, suppressing T-cell mechanosensing and cytotoxicity to drive tumor immune evasion. However, existing approaches to modulate membrane stiffness lack specificity, limiting targeted therapeutic potential. Here, we report a dual-spatially confined DNA nanowall assembly strategy to enhance tumor cell stiffness, thereby augmenting the efficacy of adoptive T-cell immunotherapy. In acidic tumor microenvironments, low-pH-triggered membrane-anchored probes (tissue-level confinement) selectively guide the construction of a DNA nanowall on the inner leaflet of the tumor cell membrane (cellular-level confinement). This biomechanical reprogramming markedly increases tumor rigidity. When combined with adoptive T-cell transfer (ACT) immunotherapy, this strategy restores T-cell mechanical force generation capacity, significantly improving tumor killing outcomes in vitro and in murine solid tumor models. Our findings underscore the critical role of biophysical signals in cancer–immune interactions, demonstrating that targeting mechanical immune checkpoints can potentiate antitumor immunity. Furthermore, these discoveries bridge biophysics and oncology, offering a paradigm for precision cancer therapy through the spatiotemporal control of cellular mechanics.

Mass spectrometry (MS)-based single- and trace-cell proteomics provides critical insights into cellular phenotypes, but widespread use is limited by the cost and complexity of advanced MS systems. We present a cost-effective, accessible workflow compatible with standard MS platforms and scalable for multiomics. The temperature-responsive agarose-based digital microfluidics (TRA-DMF) platform enables one-step sample processing, including lysis, reduction/alkylation, and digestion, in a parallel four-channel format. Unlike conventional droplet-based microfluidics or fluorescence-activated cell sorting (FACS) approaches, our DMF system ensures real-time visualization and confirmation of single-cell capture and 98.3% sample recovery, minimizing losses through nonpipetting transfer. The TRA-DMF system also overcomes the MS-incompatibility of oil-phase microfluidics, allowing high-efficiency droplet transfer to MS vials. The entire TRA-DMF for single cell proteomics (TRA-DMF-SCP) workflow is completed in ∼3 h (including single-cell capture), with seamless sample introduction into standard LC-MS/MS systems. Using a regular benchtop MS instrument Orbitrap Fusion, we identified over 4000 protein groups (PGs) from 50 293T cells with excellent reproducibility and robustness. This system offers a practical and scalable solution for trace- and single-cell proteomics and holds strong potential for integration into routine multiomics workflows.

The lysosome-targeting chimera (LYTAC) approach, especially aptamer-based LYTAC, has shown promise for the targeted degradation of secreted and membrane proteins. However, the conventional monovalent aptamer-based LYTAC has limited degradation efficiency, due to poor cellular internalization and limited serum stability. Herein, we developed a tetrahedral DNA and aptamer-based multivalent lysosome targeting chimera (TDA-MLYTAC) for targeted degradation of single or dual protein targets. Benefiting from the properties of tetrahedral DNA, TDA-MLYTAC demonstrates significant improvement in cellular uptake, precise target identification, and cellular stability. Our study proved that TDA-MLYTAC modified with trivalent triantenerrary N-acetylgalactosamine and monovalent aptamer induced liver-cell specific degradation of membrane proteins twice as efficiently as the monovalent aptamer-based LYTAC through the lysosomal pathway. The bispecific TDA-MLYTAC modified with two aptamers has great potential for inducing dual-targeted protein degradation. By rationally designing the recognition elements for receptors and proteins, it is possible to revolutionize the development of targeted protein degradation tools and lay the groundwork for receptor-mediated drug therapies.

Multivalent strategies have been widely applied in the microfluidic interface to boost the capture efficiency of target cells. However, achieving a balance between binding kinetics and thermodynamics in existing multivalent affinity interfaces remains challenging. Here, we propose a synergistic Aptamer-nanobody hetero-Multivalency Programmable magnetic fluid microfluidic chip (AMP-chip) which utilizes the combined advantages of ligands to enhance both thermodynamic and kinetic properties of the capture interface. The AMP-chip integrates two distinct noninterfering recognition molecules: one with high affinity and another with rapid binding capability, both of which are assembled onto nanomagnetic beads. This integration achieves intermolecular complementarity, effectively enhancing the binding kinetics and thermodynamic stability. We chose mutually noninterfering CD71 recognition targets, a high-affinity nanobody (NB) and a rapid-binding aptamer (XQ 2d), and fully utilized the respective advantages of these ligands to facilitate rapid and tight recognition of the CD71 receptor on target cells. By integrating a herringbone microarray into an AMP-chip to further increase the cell-ligand interaction, we significantly improved the sensitivity and accuracy of circulating nucleated red blood cell (cNRBC) isolation from the peripheral blood mononuclear cells (PBMCs) of pregnant women. Additionally, the ligands were primarily fixed to the chip by magnetic force without chemical bonding, enabling nondestructive cell release and preserving high cell viability for subsequent molecular analyses. Overall, this approach offers a novel thermodynamic-kinetic synergistic heteromultivalency interface with significant potential for clinical applications.

Extracellular vesicles (EVs) are heterogeneous lipid containers carrying complex molecular cargoes, including proteins, nucleic acids, glycans, etc. These vesicles are closely associated with specific physiological characteristics, which makes them invaluable in the detection and monitoring of various diseases. However, traditional isolation methods are often labour-intensive, inefficient, and time-consuming. In addition, single biomarker analyses are no longer accurate enough to meet diagnostic needs. Routine isolation and molecular analysis of high-purity EVs in clinical applications is even more challenging. In this review, we discuss a promising solution, microfluidic-based techniques, that combine efficient isolation and multiplex detection of EVs, to further demystify EV heterogeneity. These microfluidic-based EV multiplexing platforms will hopefully facilitate development of liquid biopsies and offer promising opportunities for personalised therapy.

Information processing is a critical topic in the digital age, as silicon-based circuits face unprecedented challenges such as data explosion, immense energy consumption, and approaching physical limits. Deoxyribonucleic acid (DNA), naturally selected as a carrier for storing and using genetic information, possesses unique advantages for information processing, which has given rise to the emerging fields of DNA computing and DNA data storage. To meet the growing practical demands, a wide variety of materials and interfaces have been introduced into DNA information processing technologies, leading to significant advancements. This review summarizes the advances in materials and interfaces that facilitate DNA computation and DNA data storage. We begin with a brief overview of the fundamental functions and principles of DNA computation and DNA data storage. Subsequently, we delve into DNA computing systems based on various materials and interfaces, including microbeads, nanomaterials, DNA nanostructures, hydrophilic-hydrophobic compartmentalization, hydrogels, metal-organic frameworks, and microfluidics. We also explore DNA data storage systems, encompassing encapsulation materials, microfluidics techniques, DNA nanostructures, and living cells. Finally, we discuss the current bottlenecks and obstacles in the fields and provide insights into potential future developments.