An in vivo platform to screen for regulators of Huntington’s disease
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Emerging insights in senescence: pathways from preclinical models to therapeutic innovations
Senescence is a crucial hallmark of ageing and a significant contributor to the pathology of age-related disorders. As committee members of the young International Cell Senescence Association (yICSA), we aim to synthesise recent advancements in the identification, characterisation, and therapeutic targeting of senescence for clinical translation. We explore novel molecular techniques that have enhanced our understanding of senescent cell heterogeneity and their roles in tissue regeneration and pathology. Additionally, we delve into in vivo models of senescence, both non-mammalian and mammalian, to highlight tools available for advancing the contextual understanding of in vivo senescence. Furthermore, we discuss innovative diagnostic tools and senotherapeutic approaches, emphasising their potential for clinical application. Future directions of senescence research are explored, underscoring the need for precise, context-specific senescence classification and the integration of advanced technologies such as machine learning, long-read sequencing, and multifunctional senoprobes and senolytics. The dual role of senescence in promoting tissue homoeostasis and contributing to chronic diseases highlights the complexity of targeting these cells for improved clinical outcomes.
Relay-projection microscopic telescopy
The fundamental trade-off between spatial resolution and imaging distance poses a significant challenge for current imaging techniques, such as those used in modern biomedical diagnosis and remote sensing. Here, we introduce a new conceptual method for imaging dynamic amplitude-phase-mixed objects, termed relay-projection microscopic telescopy (rPMT), which fundamentally challenges conventional light collection techniques by employing non-line-of-sight light collection through square-law relay-projection mechanisms. We successfully resolved tiny features measuring 2.76 μm, 22.10 μm, and 35.08 μm for objects positioned at distances of 1019.0 mm, 26.4 m, and 96.0 m, respectively, from single-shot spatial power spectrum images captured on the relay screen; these results demonstrate that the resolution capabilities of rPMT significantly surpass the Abbe diffraction limit of the 25 mm-aperture camera lens at the respective distances, achieving resolution improvement factors of 7.9, 25.4, and 58.2. The rPMT exhibits long-distance, wide-range, high-resolution imaging capabilities that exceed the diffraction limit of the camera lens and the focusing range limit, even when the objects are obscured by a scattering medium. The rPMT enables telescopic imaging from centimeters to beyond hundreds of meters with micrometer-scale resolution using simple devices, including a laser diode, a portable camera, and a diffusely reflecting whiteboard. Unlike contemporary high-resolution imaging techniques, our method does not require labeling reagents, wavefront modulation, synthetic receive aperture, or ptychography scanning, which significantly reduce the complexity of the imaging system and enhance the application practicality. This method holds particular promise for in-vivo label-free dynamic biomedical microscopic imaging diagnosis and remote surveillance of small objects.
In vivo surface-enhanced Raman scattering techniques: nanoprobes, instrumentation, and applications
Surface-enhanced Raman scattering (SERS) has emerged as a powerful tool in various biomedical applications, including in vivo imaging, diagnostics, and therapy, largely due to the development of near-infrared (NIR) active SERS substrates. This review provides a comprehensive overview of SERS-based applications in vivo, focusing on key aspects such as the design considerations for SERS nanoprobes and advancements in instrumentation. Topics covered include the development of NIR SERS substrates, Raman label compounds (RLCs), protective coatings, and the conjugation of bioligands for targeted imaging and therapy. The review also discusses microscope-based configurations such as scanning, widefield imaging, and fiber-optic setups. Recent advances in using SERS nanoprobes for in vivo sensing, diagnostics, biomolecule screening, multiplex imaging, intraoperative guidance, and multifunctional cancer therapy are highlighted. The review concludes by addressing challenges in the clinical translation of SERS nanoprobes and outlines future directions, emphasizing opportunities for advancing biomedical research and clinical applications.
Customizable virus-like particles deliver CRISPR–Cas9 ribonucleoprotein for effective ocular neovascular and Huntington’s disease gene therapy
In vivo CRISPR gene editing holds enormous potential for various diseases. Ideally, CRISPR delivery should be cell type-specific and time-restricted for optimal efficacy and safety, but customizable methods are lacking. Here we develop a cell-tropism programmable CRISPR–Cas9 ribonucleoprotein delivery system (RIDE) based on virus-like particles. The efficiency of RIDE was comparable to that of adeno-associated virus and lentiviral vectors and higher than lipid nanoparticles. RIDE could be readily reprogrammed to target dendritic cells, T cells and neurons, and significantly ameliorated the disease symptoms in both ocular neovascular and Huntington’s disease models via cell-specific gene editing. In addition, RIDE could efficiently edit the huntingtin gene in patients’ induced pluripotent stem cell-derived neurons and was tolerated in non-human primates. This study is expected to facilitate the development of in vivo CRISPR therapeutics.
A genome-wide atlas of human cell morphology
A key challenge of the modern genomics era is developing empirical data-driven representations of gene function. Here we present the first unbiased morphology-based genome-wide perturbation atlas in human cells, containing three genome-wide genotype–phenotype maps comprising CRISPR–Cas9-based knockouts of >20,000 genes in >30 million cells. Our optical pooled cell profiling platform (PERISCOPE) combines a destainable high-dimensional phenotyping panel (based on Cell Painting) with optical sequencing of molecular barcodes and a scalable open-source analysis pipeline to facilitate massively parallel screening of pooled perturbation libraries. This perturbation atlas comprises high-dimensional phenotypic profiles of individual cells with sufficient resolution to cluster thousands of human genes, reconstruct known pathways and protein–protein interaction networks, interrogate subcellular processes and identify culture media-specific responses. Using this atlas, we identify the poorly characterized disease-associated TMEM251/LYSET as a Golgi-resident transmembrane protein essential for mannose-6-phosphate-dependent trafficking of lysosomal enzymes. In sum, this perturbation atlas and screening platform represents a rich and accessible resource for connecting genes to cellular functions at scale.
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