Ryan Group

The Ryan lab uses high-throughput biology approaches to understand neurodegenerative diseases such as Parkinson’s using cellular models. We are particularly interested in the role of mitochondrial dysfunction and mitophagy in the loss of dopaminergic neurons.

3D reconstruction of i3 glutamatergic neurons (Image by Rachel Heon-Roberts)

Understanding mitophagy regulation

Mitochondrial dysfunction is a key feature of many diseases, including Parkinson’s. Genetics point to mitochondrial quality control (mitophagy) by PINK1 and PRKN as being crucial for dopaminergic neuronal health. We use iPSC-derived dopaminergic and glutamatergic neurons to understand the physiological/pathophysiological triggers of mitophagy and how Parkinson’s-causing mutations cause neuronal dysfunction (Williamson and Madureira 2023).

Using this knowledge, we can screen for, or test, novel therapeutic approaches to improve mitochondrial health. We use and have developed a number of assays for assessing mitochondrial health and mitophagy using high-content imaging and HTRF in 384-well format.

Functional Genomics

We use CRISPRi and CRISPRa as key tools to understand the effects of disease-linked gene variation on disease-biology, at scale, in collaboration with the Ward lab . We have created several CRISPRi libraries to probe the effect of gene knockdown on disease-relevant biology including both mitochondrial and lysosomal dysfunction. We can also use these tools to understand genetic interactions and phenocopy disease-causing mutations in various cell types across neurodegeneration (Franks and Heon-Roberts 2024 ).

Identifying new therapeutic approaches for Parkinson’s

We use a range of proteomics and high-content imaging approaches to identify novel proteins and pathways in neurodegeneration.

We use a number of proteomics approaches, including post-translational proteomics (PTMomics) to identify novel cell biology and potential therapeutic approaches in models of neurodegeneration (Bogetofte 2023 ). This work is carried out in close collaboration with Helle Bogetofte and the Larsen Lab at the University of Southern Denmark.

We also make use of high-throughput screening facilities to enable phenotypic screening of iPSC-derived neuronal models to identify novel therapeutics and validate omics datasets. To enable this, we work with well-characterised small molecule or CRISPRi/a libraries to assess mitochondrial and lysosomal phenotypes. To better understand neuronal biology, and embrace the complexity of genetic or chemical perturbations, we increasingly use multiplexed analyses such as Cell Painting.