ALS is the most common form of motor neuron disease. We have multiple long-standing projects investigating the underlying pathological mechanisms, ranging from analysis of splicing defects to the study of impaired proteostasis.

Kennedy’s disease, also known as spinal and bulbar muscular atrophy (SBMA) is a type of late-onset motor neuron disease caused by a repeat expansion in the gene encoding androgen receptor. By combining insights from the clinic and from experiments on disease models, we are improving understanding of this devastating disease.

Many neurodegenerative disorders involve the dysregulation of splicing and polyadenylation. We are actively investigating splicing and polyadenylation defects in ALS and related neurodegenerative diseases by searching for novel aberrant splicing events and validating their importance via an array of experimental approaches.

The loss of neuronal proteostasis is a hallmark of many neurodegenerative disorders. By combining techniques biochemical techniques such as ribosome proximity ligation and puromycinylation assays with high throughput approaches including ribosome profiling and mass spectrometry, we are improving understanding of how proteostasis is impaired in these disorders.

We are interested in the regulation of RNA production and degradation. We have elucidated important post-transcription regulatory mechanisms for controlling gene expression and we are actively studying the dysregulation RNA decay in disease models using techniques such as 4sU pulse chase assays.

We are actively developing new targeted therapeutics to treat motor neuron disease. We are exploring a wide range of therapeutic approaches, from more conventional technologies such as anti-sense oligonucleotide therapies, to novel targeted medicine approaches to improve efficacy and reduce side-effects.

Biomarkers are extremely important, both for accurately diagnosing patients, and for monitoring the efficacy of novel treatments during clinical trials. We are working to establish new biomarkers to help progress this field.

The majority of motor neuron disease cases have no known genetic cause. We are working to understand the genetics that underpin these diseases, which help elucidate novel disease mechanisms that may be of broad relevance to neurodegenerative disorders.

We routinely produce high-throughput sequencing libraries to enable transcriptome-wide interrogation of neurodegeneration-related defects. In addition to standard RNA-sequencing experiments, we perform:

• iCLIP (protein-RNA interactions)

• 4sU pulse chase (RNA stability)

• Ribosome profiling (Protein synthesis)

• Mutagenesis screens (Drug development)

Our team of bioinformaticians routinely produce custom analysis pipelines for the study of neurodegeneration-related aberrancies and the discovery of novel disease mechanisms. These pipelines are applied to both our own in-house experiments and to published datasets. Additionally, when necessary we produce our own custom software and algorithms. Our github is available here https://github.com/frattalab/

Microscopy is essential for much of our work. We have access to:

• Multiple confocal microscopes

• Automated imaging platforms for time-course experiments

• Super-resolution microscopes

Additionally, we have been involved in the development of AI-based software for classifying defects in neuromuscular junctions

Much of research involves mammalian cell culture. We routinely work with:

• iPSC-derived neurons

• Primary neurons

• Transformed mouse embryonic fibroblasts derived from our murine disease models

• Neuron-like cell lines (eg. SH-SY5Y)

We also work with various animal models of disease. Some of our key mouse models include:

• Delta-14 FUS (ALS)

• AR-100x CAG (SBMA)

Although we aim to use cellular models where possible, there are circumstances in which animal models are essential to provide deeper insight into disease mechanisms.

To complement our high-throughput experiments we perform targeted experiments for validation or to answer questions that cannot otherwise be addressed. In addition to standard molecular biology techniques, our studies often involve:

• RT-PCR analysis of splicing patterns

• PLA-based validation of protein-protein interactions or protein synthesis

• Biophysical characterization of molecular interactions (eg. isothermal titration calorimetry)