Subcellular visualization: Organelle-specific targeted drug delivery and discovery

Shao, Xintian; Meng, Chen; Song, Wenjing; Zhang, Tao; Chen, Qixin

doi:10.1016/j.addr.2023.114977

Cited by 25 publications

(4 citation statements)

References 258 publications

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“…Selecting optimal targeting ligands through understanding subcellular processes associated with productive delivery will enable improved design of nano-delivery systems. Further refinement of drug delivery systems to dial-in subcellular function could be critical to realize next-generation systems with improved therapeutic index [ 64 , 65 ]. Furthermore, the basic assay design described here could be further expanded-on to answer more complex questions regarding trafficking and function of targeted NPs and further guide NP design.…”

Section: Resultsmentioning

confidence: 99%

A high-throughput lysosome trafficking assay guides ligand selection and elucidates differences in CD22-targeted nanodelivery

Vaughan,

Est-Witte,

Dockery

et al. 2024

Science and Technology of Advanced Materials

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Targeted nanoparticles offer potential to selectively deliver therapeutics to cells; however, their subcellular fate following endocytosis must be understood to properly design mechanisms of drug release. Here we describe a nanoparticle platform and associated cell-based assay to observe lysosome trafficking of targeted nanoparticles in live cells. The nanoparticle platform utilizes two fluorescent dyes loaded onto PEG-poly(glutamic acid) and PEG-poly(Lysine) block co-polymers that also comprise azide reactive handles on PEG termini to attach antibody-based targeting ligands. Fluorophores were selected to be pH-sensitive (pHrodo Red) or pH-insensitive (Alexafluor 488) to report when nanoparticles enter low pH lysosomes. Dye-labelled block co-polymers were further assembled into polyion complex micelle nanoparticles and crosslinked through amide bond formation to form stable nano-scaffolds for ligand attachment. Cell binding and lysosome trafficking was determined in live cells by fluorescence imaging in 96-well plates and quantification of red- and green-fluorescence signals over time. The platform and assay was validated for selection of optimal antibody-derived targeting ligands directed towards CD22 for nanoparticle delivery. Kinetic analysis of uptake and lysosome trafficking indicated differences between ligand types and the ligand with the highest lysosome trafficking efficiency translated into effective DNA delivery with nanoparticles bearing the optimal ligand.

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Section: Resultsmentioning

confidence: 99%

A high-throughput lysosome trafficking assay guides ligand selection and elucidates differences in CD22-targeted nanodelivery

Vaughan,

Est-Witte,

Dockery

et al. 2024

Science and Technology of Advanced Materials

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“…We also used super-resolution imaging to investigate the interactions between lysosomes and lipid droplets [ 12 , 30 , 31 ] upon treatment with FITC-MOS-1 ( Fig. S24 ).…”

Section: Resultsmentioning

confidence: 99%

“…Several modern research strategies, including high-throughput screening techniques [ 9 ], seek to identify diseases and indications for bioactive components of NPs. Technologies that combine fluorescent labeling with live-cell imaging [ [10] , [11] , [12] ] represent attractive cell-based strategies for use in NP-related research. Such studies involve real-time tracking of drug distribution within cells and subcellular compartments.…”

Section: Introductionmentioning

confidence: 99%

In situ visualization of the cellular uptake and sub-cellular distribution of mussel oligosaccharides

Yu,

Shao,

Shao

et al. 2024

Journal of Pharmaceutical Analysis

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“…3 So far, many efforts have been devoted to improving the pharmacokinetics, biodistribution, and cellular internalization efficiency of NRs. 4 However, considering the complex tumor microenvironment, the intracellular distribution and location of NRs are also very important to their clinical efficacy, with very different therapeutic outcomes for NRs adsorbed on the cell surface, internalized into the cell, or subcellular location. NRs need to overcome multiple barriers to induce effective cancer cell deaths, which include the cellular membrane, endo/ lysosomal degradation, cytoplasmic transportation, and specific organelle targeting.…”

Section: Introductionmentioning

confidence: 99%

Quantitative Three-Dimensional Imaging Analysis of HfO₂ Nanoparticles in Single Cells via Deep Learning Aided X-ray Nano-Computed Tomography

Xi,

Yao,

Zhang

et al. 2024

ACS Nano

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It is crucial for understanding mechanisms of drug action to quantify the three-dimensional (3D) drug distribution within a single cell at nanoscale resolution. Yet it remains a great challenge due to limited lateral resolution, detection sensitivities, and reconstruction problems. The preferable method is using X-ray nano-computed tomography (Nano-CT) to observe and analyze drug distribution within cells, but it is time-consuming, requiring specialized expertise, and often subjective, particularly with ultrasmall metal nanoparticles (NPs). Furthermore, the accuracy of batch data analysis through conventional processing methods remains uncertain. In this study, we used radioenhancer ultrasmall HfO 2 nanoparticles as a model to develop a modular and automated deep learning aided Nano-CT method for the localization quantitative analysis of ultrasmall metal NPs uptake in cancer cells. We have established an ultrasmall objects segmentation method for 3D Nano-CT images in single cells, which can highly sensitively analyze minute NPs and even ultrasmall NPs in single cells. We also constructed a localization quantitative analysis method, which may accurately segment the intracellularly bioavailable particles from those of the extracellular space and intracellular components and NPs. The high bioavailability of HfO 2 NPs in tumor cells from deeper penetration in tumor tissue and higher tumor intracellular uptake provide mechanistic insight into HfO 2 NPs as advanced radioenhancers in the combination of quantitative subcellular image analysis with the therapeutic effects of NPs on 3D tumor spheroids and breast cancer. Our findings unveil the substantial uptake rate and subcellular quantification of HfO 2 NPs by the human breast cancer cell line (MCF-7). This revelation explicates the notable efficacy and safety profile of HfO 2 NPs in tumor treatment. These findings demonstrate that this 3D imaging technique promoted by the deep learning algorithm has the potential to provide localization quantitative information about the 3D distributions of specific molecules at the nanoscale level. This study provides an approach for exploring the subcellular quantitative analysis of NPs in single cells, offering a valuable quantitative imaging tool for minute amounts or ultrasmall NPs.

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Subcellular visualization: Organelle-specific targeted drug delivery and discovery

Cited by 25 publications

References 258 publications

A high-throughput lysosome trafficking assay guides ligand selection and elucidates differences in CD22-targeted nanodelivery

A high-throughput lysosome trafficking assay guides ligand selection and elucidates differences in CD22-targeted nanodelivery

In situ visualization of the cellular uptake and sub-cellular distribution of mussel oligosaccharides

Quantitative Three-Dimensional Imaging Analysis of HfO₂ Nanoparticles in Single Cells via Deep Learning Aided X-ray Nano-Computed Tomography

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Subcellular visualization: Organelle-specific targeted drug delivery and discovery

Cited by 25 publications

References 258 publications

A high-throughput lysosome trafficking assay guides ligand selection and elucidates differences in CD22-targeted nanodelivery

A high-throughput lysosome trafficking assay guides ligand selection and elucidates differences in CD22-targeted nanodelivery

In situ visualization of the cellular uptake and sub-cellular distribution of mussel oligosaccharides

Quantitative Three-Dimensional Imaging Analysis of HfO2 Nanoparticles in Single Cells via Deep Learning Aided X-ray Nano-Computed Tomography

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Product

Resources

About

Quantitative Three-Dimensional Imaging Analysis of HfO₂ Nanoparticles in Single Cells via Deep Learning Aided X-ray Nano-Computed Tomography