Gene Therapy Vectors Assessment

Newcells Retinal Model

Accelerate your lead compound selection by understanding their mode of action in functional retinal tissue

Recapitulate the architecture and function of the human retina

Evaluate gene therapy vectors and novel drugs for retinal diseases in vitro

Accelerate drug discovery by replacing animal experiments

Accelerated in vitro preclinical evaluation of viral vectors for retinal gene therapy development

Newcells retinal organoids and RPE models are valuable tools for in vitro preclinical selection of new retinal gene therapy vectors. They allow rapid in vitro evaluation of the best combination of vector capsid, promoter, and transgene. They also enable initial safety and efficacy testing to be carried out using the exact same promoter or gene as those to be used in clinical trials. Our retinal models have been validated for their use in screening and selection of optimal adeno-associated viral (AAV) vectors with increased photoreceptor tropism for retinal gene therapy.

Service outputs

Evaluation of AAV vectors transduction efficiency in iPSC-derived retinal organoids or RPE cells
Determination of AAV vector cell tropism and transduction of photoreceptor-like cells in retinal organoids
Cell viability assessment post-transduction

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Localization and distribution of Müller glia cells (CRALBP, red) in retinal organoids. Nuclear DAPI staining (blue).

Gene therapy vector tropism

Viral vector optimisation

In vitro evaluation

Outputs

Transduction efficiency of viral vector
In vitro safety prediction
Cell viability assessment
Therapeutic efficacy
Cell tropism identification in retinal organoids

Models

Human retinal organoids
Human retinal pigment epithelial (RPE) cells
- derived from heathy donor
- derived from patient or CRISPR/Cas9 gene-edited iPSCs supplied by the client
- derived from patient or CRISPR/Cas9 gene-edited PBMCs or fibroblasts supplied by the client

Timeline

2-6 months

How to use our gene therapy vector assessment service?

Outsource your experiments to us to obtain robust data

Take advantage of our expertise: our team designs the experiments with you to help you select the most effective gene therapy vector for your target cells. A fast and reliable service allows for rapid in vitro screening of novel viral vectors in a human model. The data is validated and supports IND applications. All bespoke projects are carried out in our state-of-the-art UK facilities.

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Generation of Patient-derived organoids and RPE as part of the service

Make use of our complete service including the generation of organoids and RPE from your patient-derived or gene-edited iPSCs, PBMCs or fibroblasts, feasibility studies, experimental design, testing of viral vectors and generation of reliable data for regulatory submissions.

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AAV vector evaluation in vitro for retinal gene therapy Close Open

Newcells’ retinal organoids have shown their potential to be used as an informative in vitro model for the evaluation of new retinal gene therapy vectors for retinal diseases.

Our organoids have been used to screen AAV gene therapy vectors in a collaboration with a team at the University of Oxford. This study confirmed robust and efficient transduction of human photoreceptor-like cells by AAV vectors highlighting improved transduction capability of AAV2 7m8 in retinal organoids when using the ubiquitous CAG promoter. With several cell types present within the organoids, the experiments were able to specifically show the targeted cell types, demonstrating that a CAG-driven transgene transduced a broad range of cell types, while GRK1-driven transgenes show a more restricted photoreceptor-specific expression (A). The work also demonstrated the viability of the retinal organoids were not affected by AAV transduction (B).

(A) Live cell imaging of reporter gene expression up to 27 days post-transduction. Arrows indicate the areas where the onset of reporter gene expression first appeared, allowing evaluation of which cell types each AAV vector transduced preferentially.

(B) Retinal organoid viability following transduction by AAV. ATP assay was assessed as an indicator of viability at 27 days post-transduction with AAV vectors. Data analysed using a one-way analysis of variance indicated no significant influence of AAV treatment on ATP levels and viability (F=1.107, P =0.3908) relative to control retinal organoids.

These studies can be carried out in both our retinal organoids and RPE in vitro models to allow rapid evaluation of new retinal gene therapy vectors.

Publication

Tropism of AAV Vectors in Photoreceptor-Like Cells of Human iPSC-Derived Retinal Organoids

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Transduction efficiency in RPE Close Open

Dose- and time- dependent increase in GFP signal in AAV-transduced RPE. Percentage of GFP-positive cells over 28 days post transduction with AAV5-CAG-EGFP:WPR at two different doses. Scale: 150 µm.

Service overview Close Open

Newcells provides a custom service evaluating AAV vectors in our laboratories. With our regular supply of tissue, our projects timelines are short. The robust data generated by our scientific experts will guide you in confidence for key decision-making steps during drug development.

An example of viral vector testing service includes a set of assays to assess transduction efficiency, cell tropism and post-transduction cell viability.

Assay Design
Models	Retinal organoids with photoreceptors (cone and rod), retinal ganglion cells, horizontal cells and amacrine cells. Retina pigment epithelium (RPE) cells (2D cobblestones monolayer).
Assay format	96-well plates (retinal organoids)
Assay format	24-well plates (RPE)
Species	Human
Assay readout	Cell viability assay (ATP depletion assay, LDH release and microscopy)
	Qualitative immunofluorescence with cell specific markers & cell death markers
	AAV transduction efficiency
	Therapeutic efficacy
Time points and replicates	Post-transduction (typically up to 28 days)
Time points and replicates	Triplicates per vector and per concentration

Models to choose from for this service

Retinal organoids

The retinal organoids are iPSC-derived, and they recapitulate the complex structure of the human retina with laminar cell organisation mimicking embryonic development. They contain the outer photoreceptor segment of the retina that responds to light.

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Retinal pigment epithelium cells (RPE)

A functional 2D in vitro model of retinal pigment epithelial cells generated from human iPSCs recapitulating phagocytosis of photoreceptor outer segments. The RPE cells are pigmented and displays typical cobblestone morphology.

Image of RPE model — RPE cells displaying cobblestone morphology. Cells were immunolabeled with tight-junction ZO-1 marker (shown in green) and co-stained with nuclei marker, Hoechst (shown in blue).

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Other services with retina tissues

Disease Modelling

We generate models of retinopathies from patient-derived or gene-edited iPSCs, PBMCs or fibroblasts supplied by our clients.

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Retinal toxicity evaluation

High throughput in vitro drug screening for retinal safety/toxicity and efficacy testing using validated human retinal organoids.

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Is this the service for you? Speak to one of our experts about your requirements.

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Gene Therapy Vectors Assessment