In situ-formable, dynamic crosslinked poly(ethylene glycol) carrier for localized adeno-associated virus infection and reduced off-target effects

Kato, Motoi; Ishikawa, Shohei; Shen, Qi; Du, Zhao‐Jiang; Katashima, Takuya; Naito, Mitsuru; Numahata, Takao; Okazaki, Mutsumi; Sakai, Takamasa; Kurita, Masanori

doi:10.1038/s42003-023-04851-w

Cited by 8 publications

(7 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…AAVs encoding green fluorescent protein with a nuclear localization signal (GFPNLS) were used to assess gene transduction efficiency because nucleus-localized fluorescence facilitates distinguishing gene expression from background signals [7]. KCs cultured on feeder cells form colonies with a defined edge [2][20].…”

Section: Resultsmentioning

confidence: 99%

“…In our experiences with repeated production of AAVs with various capsids, promoters, and genes [2][7][23], AAV production using the same procedure is substantially affected by the type of capsid, promoter, and gene. We examined AAV production of various types of capsids and promoters, and confirmed significant differences, suggesting that AAV production is a practical reason for selecting AAV types, especially when used for experiments.…”

Section: Resultsmentioning

confidence: 99%

“…However, most AAV vectors cause non-specific effects [24]. In this context, the specificity of gene expression at target tissues, minimizing off-target effects, is important for in vivo applications [7]. Changing the tropism of an AAV to a specific cell type may affect its interaction with other cell types.…”

Section: Discussionmentioning

confidence: 99%

“…Adeno-associated viruses (AAVs) are useful vectors for in vivo gene transduction and are clinically used as replacement therapies to treat diseases such as lipoprotein lipase deficiency, spinal muscular atrophy, retinal dystrophy, and hemophilia [5]. Recently, they have become promising vectors for the development of potent therapeutics targeting localized morbidities [2][6][7] and therapeutic in vivo genome editing [8].…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Optimization of an adeno-associated viral vector for epidermal keratinocytesin vitroandin vivo

Shen,

Suga,

Moriwaki

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

Background: Local gene therapies, including in vivo genome editing, are highly anticipated for the treatment of genetic diseases in skin, especially the epidermis. While the adeno-associated virus (AAV) is a potent vector for in vivo gene delivery, the lack of efficient gene delivery methods has limited its clinical applications. Objective: To optimize the AAV gene delivery system with higher gene delivery efficiency and specificity for epidermis and keratinocytes (KCs), using AAV capsid and promoter engineering technologies. Methods: AAV variants with mutations in residues reported to be critical to determine the tropism of AAV2 for KCs were generated by site-directed mutagenesis of AAVDJ. The infection efficiency and specificity for KCs of these variants were compared with those of previously reported AAVs considered to be suitable for gene delivery to KCs in vitro and in vivo. Additionally, we generated an epidermis-specific promoter using the most recent short-core promoter and compared its specificity with existing promoters. Results: A novel AAVDJ variant capsid termed AAVDJK2 was superior to the existing AAVs in terms of gene transduction efficiency and specificity for epidermis and KCs in vitro and in vivo. A novel tissue-specific promoter, termed the K14 SCP3 promoter, was superior to the existing promoters in terms of gene transduction efficiency and specificity for KCs. Conclusion: The combination of the AAVDJK2 capsid and K14 SCP3 promoter improves gene delivery to epidermis in vivo and KCs in vitro. The novel AAV system may benefit experimental research and development of new epidermis-targeted gene therapies.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Optimization of an adeno-associated viral vector for epidermal keratinocytesin vitroandin vivo

Shen,

Suga,

Moriwaki

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Based on the prior section, there is a clear necessity for model materials with precisely controlled network structures to elucidate molecular mechanisms. Recently, we developed model transient networks utilizing tetrafunctional precursors (Tetra-PEG slimes) by employing dynamic covalent bonds between 4-carboxy-3-fluorophenylboronic acid (FPBA) and glucono-δ-lactone (GDL) as reversible crosslinks [30,[68][69][70]. Due to the temporal characteristics of the crosslinkers, the Tetra-PEG slime exhibits the flow behavior in a long time limit, classified into transient networks.…”

Section: Relationships Between Translational Diffusion and Viscoelast...mentioning

confidence: 99%

Probing the Molecular Mechanism of Viscoelastic Relaxation in Transient Networks

Michida,

Chung,

Katashima

2023

Gels

Self Cite

View full text Add to dashboard Cite

Hydrogels, which have polymer networks through supramolecular and reversible interactions, exhibit various mechanical responsibilities to its surroundings. The influence of the reversible bonds on a hydrogel’s macroscopic properties, such as viscoelasticity and dynamics, is not fully understood, preventing further innovative material development. To understand the relationships between the mechanical properties and molecular structures, it is required to clarify the molecular understanding of the networks solely crosslinked by reversible interactions, termed “transient networks”. This review introduces our recent progress on the studies on the molecular mechanism of viscoelasticity in transient networks using multiple methods and model materials. Based on the combination of the viscoelasticity and diffusion measurements, the viscoelastic relaxation of transient networks does not undergo the diffusion of polymers, which is not explained by the framework of conventional molecular models for the viscoelasticity of polymers. Then, we show the results of the comparison between the viscoelastic relaxation and binding dynamics of reversible bonds. Viscoelastic relaxation is primarily affected by “dissociation dynamics of the bonds” and “network structures”. These results are explained in the framework that the backbone, which is composed of essential chains supporting the stress, is broken by multiple dissociation events. This understanding of molecular dynamics in viscoelasticity will provide the foundation for designing transient networks.

show abstract

Viral Vector Eluting Lenses for Single‐Step Targeted Expression of Genetically‐Encoded Activity Sensors for in Vivo Microendoscopic Calcium Imaging

Jons,

Cheng,

Dong

et al. 2024

Macromolecular Bioscience

View full text Add to dashboard Cite

Optical methods for studying the brain offer powerful approaches for understanding how neural activity underlies complex behavior. These methods typically rely on genetically encoded sensors and actuators to monitor and control neural activity. For microendoscopic calcium imaging, injection of a virus followed by implantation of a lens probe is required to express a calcium sensor and enable optical access to the target brain region. This two‐step process poses several challenges, chief among them being the risks associated with mistargeting and/or misalignment between virus expression zone, lens probe and target brain region. Here, an adeno‐associated virus (AAV)‐eluting polymer coating is engineered for gradient refractive index (GRIN) lenses enabling the expression of a genetically encoded calcium indicator (GCaMP) directly within the brain region of interest upon implantation of the lens. This approach requires only one surgical step and guarantees alignment between GCaMP expression and lens in the brain. Additionally, the slow virus release from these coatings increases the working time for surgical implantation, expanding the brain regions and species amenable to this approach. These enhanced capabilities should accelerate neuroscience research utilizing optical methods and advance the understanding of the neural circuit mechanisms underlying brain function and behavior in health and disease.

show abstract

In situ-formable, dynamic crosslinked poly(ethylene glycol) carrier for localized adeno-associated virus infection and reduced off-target effects

Cited by 8 publications

References 33 publications

Optimization of an adeno-associated viral vector for epidermal keratinocytesin vitroandin vivo

Optimization of an adeno-associated viral vector for epidermal keratinocytesin vitroandin vivo

Probing the Molecular Mechanism of Viscoelastic Relaxation in Transient Networks

Viral Vector Eluting Lenses for Single‐Step Targeted Expression of Genetically‐Encoded Activity Sensors for in Vivo Microendoscopic Calcium Imaging

Contact Info

Product

Resources

About