Disease Modelling

Following the generation of retinal organoids from RP Type 11 patients with a mutation in pre-mRNA processing factor 31 (PRPF31), the mechanism of retinal dysfunction associated with the disease has been elucidated. Large-scale transcriptome analyses identified cell type-specific and patient-specific mis-splicing of PRPF31 target genes affected by PRPF31 mutations, providing unprecedented molecular characterisation of splicing-factor RP clinical phenotypes. The cellular defects unravelled in this work included dysfunctional RPE, disrupted cilia morphology in photoreceptors, progressive cellular degeneration, and cellular stress, as shown by phenotypic rescue. This work was the first demonstration of the cellular phenotypes associated with RP using patient derived organoids or RPEs.

This work and expertise was established in Newcells Biotech co-founder’s lab Prof. Lako, Professor of Stem Cell Sciences, Biosciences Institute, Faculty of Medical Sciences,
Newcastle University

Buskin A, et al., Disrupted alternative splicing for genes implicated in splicing and ciliogenesis causes PRPF31 retinitis pigmentosa. Nat Commun. 2018 Oct 12;9(1):4234. doi: 10.1038/s41467-018-06448-y.

iPSC technology allows the engineering of patient-specific retinal organoids and RPE, offering the chance to investigate disease mechanisms and evaluate novel therapeutics. As our models are derived from human iPSCs, patient-specific gene-edited lines can be engineered allowing direct comparison between WT and mutants.

The retinal organoids are obtained through a carefully controlled differentiation process recapitulating the timeline of embryonic retinogenesis. At day 150, the organoids comprise all key cell types and are functional, allowing the testing of new compounds by simply adding them to the plate. The cell structure integrity and the gene expression profiles of key markers for the main cell types, such as photoreceptors, is assessed. We also perform qualitative imaging and microscopy to provide the full picture of the drug profile.

RPE and retinal organoids can be generated from the parental iPSC line allowing complementary analyses of the retinal tissue and epithelium.

One of the main advantages of our models for retinal disease modelling is their human origin, allowing us to generate robust and predictive data for transitional and clinical studies.

Age related Macular degeneration (AMD) disease mechanisms are largely unknown; thus patient-derived retinal organoids and RPE are very valuable tools for disease modelling. Complement and ARMS/HTRA genes are known to increase the risk of AMD, but the role of the other genes and how they increase the risk of being affected by AMD is not yet fully understood. Newcells have generated iPSC-derived RPE from individuals with low-risk and high-risk CFH (Y402H) complement polymorphisms to model AMD. Compared to low-risk individuals, the high-risk iPSC-derived RPE cells show characteristics typical of AMD including cellular, structural, and functional deficiencies associated with inflammation, cellular stress, accumulation of lipid droplets and deposits that mimic AMD. These studies can be carried out in both our retinal organoids and companion RPE.

Our service provides insights into the molecular basis of inherited retinopathies

We can also test the efficacy of novel treatments in vitro to accelerate lead development or model phenotypic rescue. Finally, we can also access and compare the safety of new drugs or viral vectors on both healthy and patient-specific derived retinal tissues.

Our service is fully customised as we generate the patient-specific lines on your behalf, or we work with your mutated iPSC lines if they are available. We will then differentiate them into organoids and RPE. For incoming lines, we first require a feasibility study.

Our differentiation protocol spans 150 to 210 days to mimic embryonic retinogenesis. Our project timelines however, are short as our streamlined manufacturing process releases a regular supply of tissue. The robust data generated by our scientific experts will guide you in confidence for key decision-making steps during drug development.

An example of disease modelling packages includes a set of assays to assess cell viability, photoreceptor or RPE functionality and degeneration, key marker expression and localisation. We can also assess the safety of new chemical drugs or new viral vectors such as AAV.