Hepatic stellate cell reprogramming via exosome-mediated CRISPR/dCas9-VP64 delivery

Luo, Nianan; Li, Jiangbin; Chen, Yafeng; Xu, Yan; Wei, Yu; Lu, Jianguo; Dong, Rui

doi:10.1080/10717544.2020.1850917

Cited by 42 publications

(35 citation statements)

References 37 publications

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“…CRISPR/dCAs9/VP64 in EVs from transfected AML12 mouse hepatocytes was delivered in vitro to mouse HSC in which expression of dCas9 , E-cadherin , and hepatocyte nuclear factor-4α ( HNF-4α ) was increased and expression of αSMA and collagen 1 was decreased, while i.v. administration of the fluorescently-tagged EVs resulted in their localization to mouse liver and were proposed as a fibrosis therapy [ 292 ].…”

Section: Hepatic Fibrosismentioning

confidence: 99%

Extracellular Vesicles in Organ Fibrosis: Mechanisms, Therapies, and Diagnostics

Brigstock

2021

Cells

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Fibrosis is the unrelenting deposition of excessively large amounts of insoluble interstitial collagen due to profound matrigenic activities of wound-associated myofibroblasts during chronic injury in diverse tissues and organs. It is a highly debilitating pathology that affects millions of people globally and leads to decreased function of vital organs and increased risk of cancer and end-stage organ disease. Extracellular vesicles (EVs) produced within the chronic wound environment have emerged as important vehicles for conveying pro-fibrotic signals between many of the cell types involved in driving the fibrotic response. On the other hand, EVs from sources such as stem cells, uninjured parenchymal cells, and circulation have in vitro and in vivo anti-fibrotic activities that have provided novel and much-needed therapeutic options. Finally, EVs in body fluids of fibrotic individuals contain cargo components that may have utility as fibrosis biomarkers, which could circumvent current obstacles to fibrosis measurement in the clinic, allowing fibrosis stage, progression, or regression to be determined in a manner that is accurate, safe, minimally-invasive, and conducive to repetitive testing. This review highlights the rapid and recent progress in our understanding of EV-mediated fibrotic pathogenesis, anti-fibrotic therapy, and fibrosis staging in the lung, kidney, heart, liver, pancreas, and skin.

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Section: Hepatic Fibrosismentioning

confidence: 99%

Extracellular Vesicles in Organ Fibrosis: Mechanisms, Therapies, and Diagnostics

Brigstock

2021

Cells

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“…While classified as EV therapies, such studies are more often comprised of a variety of secreted components (i.e., secretome containing soluble factors and EVs) than purified EVs ( Table 1 ). Various terminology is used in the field ( Thery et al, 2018 ), including “extracellular vesicles,” “exosomes,” “secretomes,” “nanoparticles,” or components “enriched in extracellular vesicles.” Regardless of terminology or composition, these therapies utilize the functional capacity EVs/secreted components have to mediate a recipient-cell response through the delivery of cargo including siRNAs ( Shtam et al, 2013 ), miRNAs ( Li L. et al, 2019 ), proteins ( Garaeva et al, 2021 ), small molecule drugs ( Tian et al, 2014 ), and molecular toolkits ( Ye et al, 2020 ; Luo et al, 2021 ; Yao X. et al, 2021 ).…”

Section: Current Developments In Ev-based Therapeuticsmentioning

confidence: 99%

Development of Extracellular Vesicle Therapeutics: Challenges, Considerations, and Opportunities

Claridge

Lozano

Poh

et al. 2021

Front. Cell Dev. Biol.

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Extracellular vesicles (EVs) hold great promise as therapeutic modalities due to their endogenous characteristics, however, further bioengineering refinement is required to address clinical and commercial limitations. Clinical applications of EV-based therapeutics are being trialed in immunomodulation, tissue regeneration and recovery, and as delivery vectors for combination therapies. Native/biological EVs possess diverse endogenous properties that offer stability and facilitate crossing of biological barriers for delivery of molecular cargo to cells, acting as a form of intercellular communication to regulate function and phenotype. Moreover, EVs are important components of paracrine signaling in stem/progenitor cell-based therapies, are employed as standalone therapies, and can be used as a drug delivery system. Despite remarkable utility of native/biological EVs, they can be improved using bio/engineering approaches to further therapeutic potential. EVs can be engineered to harbor specific pharmaceutical content, enhance their stability, and modify surface epitopes for improved tropism and targeting to cells and tissues in vivo. Limitations currently challenging the full realization of their therapeutic utility include scalability and standardization of generation, molecular characterization for design and regulation, therapeutic potency assessment, and targeted delivery. The fields’ utilization of advanced technologies (imaging, quantitative analyses, multi-omics, labeling/live-cell reporters), and utility of biocompatible natural sources for producing EVs (plants, bacteria, milk) will play an important role in overcoming these limitations. Advancements in EV engineering methodologies and design will facilitate the development of EV-based therapeutics, revolutionizing the current pharmaceutical landscape.

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“…Luo et al [ 92 ] tried to use exosomes carrying CRISPR/Cas9 components for anti-fibrotic treatment of the murine liver. They used an inactivated variant of Cas9 (dCas9) fused to the VP64 transactivator domain for target site specific activation of gene expression.…”

Section: Producer Cell Based Exosome Engineeringmentioning

confidence: 99%

“…Exosomes from producer cells were isolated and incubated with mouse hepatic stellate cells that are associated with fibrosis. Induced expression of HNF4a caused a phenotypic change of fibrosis associated hepatic stellate cells into hepatocyte-like cells [ 92 ].…”

Section: Producer Cell Based Exosome Engineeringmentioning

confidence: 99%

CRISPR/Cas9: Principle, Applications, and Delivery through Extracellular Vesicles

Horodecka

Duechler

2021

IJMS

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The establishment of CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9) technology for eukaryotic gene editing opened up new avenues not only for the analysis of gene function but also for therapeutic interventions. While the original methodology allowed for targeted gene disruption, recent technological advancements yielded a rich assortment of tools to modify genes and gene expression in various ways. Currently, clinical applications of this technology fell short of expectations mainly due to problems with the efficient and safe delivery of CRISPR/Cas9 components to living organisms. The targeted in vivo delivery of therapeutic nucleic acids and proteins remain technically challenging and further limitations emerge, for instance, by unwanted off-target effects, immune reactions, toxicity, or rapid degradation of the transfer vehicles. One approach that might overcome many of these limitations employs extracellular vesicles as intercellular delivery devices. In this review, we first introduce the CRISPR/Cas9 system and its latest advancements, outline major applications, and summarize the current state of the art technology using exosomes or microvesicles for transporting CRISPR/Cas9 constituents into eukaryotic cells.

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Hepatic stellate cell reprogramming via exosome-mediated CRISPR/dCas9-VP64 delivery

Cited by 42 publications

References 37 publications

Extracellular Vesicles in Organ Fibrosis: Mechanisms, Therapies, and Diagnostics

Extracellular Vesicles in Organ Fibrosis: Mechanisms, Therapies, and Diagnostics

Development of Extracellular Vesicle Therapeutics: Challenges, Considerations, and Opportunities

CRISPR/Cas9: Principle, Applications, and Delivery through Extracellular Vesicles

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