Gene Therapy Vector Assessment

Retina models for rapid evaluation of viral vectors

Accelerate your viral vector selection using functional RPE and retinal organoid models

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

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

Newcells iPSC derived 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 and also enable initial safety and efficacy testing to be carried out using the exact same human promoter or gene as those to be used in clinical trials. Our retinal models provide predictive data and 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
Analysis of AAV vector cell tropism and transduction of photoreceptor-like cells in retinal organoids and RPE
Post-transduction cell viability assessment for safety prediction

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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

Assays

Transduction efficiency of viral vector by live fluoresence imaging (or flow cytometry for RPE)
Cell tropism identification in retinal organoids by immunofluorescence
Cell viability assessment for safety prediction
Therapy efficacy

Models

Human retinal organoids (RPE)
Human retinal pigment epithelial cells (RPE)
Models are derived from WT iPSCs or patient/gene-edited iPSCs 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. This fast and reliable service allows for rapid in vitro screening of novel viral vectors in human models. 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 models

Make use of our complete service including the generation of organoids and RPE from your patient-derived or gene-edited iPSCs. We will carry out a full feasibility study and experimental design prior to carrying out further testing of viral vectors and generation of reliable data for regulatory submissions.

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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
Viral vector Assessment Assay readout	AAV transduction efficiency by live fluorescence imaging or flow cytometry
	Cell tropism identification by immunofluorescence with cell specific markers for retinal organoids
	Cell viability assay for safety prediction
	Therapy efficacy
Time points	Post-transduction (typically up to 28 days)
Replicates	Triplicates per vector and per concentration

AAV vector evaluation in vitro for retinal gene therapy Close Open

Newcells’ retinal organoids have shown their potential as an 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 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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RPE Transduction efficiceny 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.

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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CONTACT US

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

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Gene Therapy Vector Assessment