Restoring Autophagy, Reversing Disease with Jess

Erasing the Signs of Cellular Damage

No other cell biological process is as strongly linked to healthy lifespan extension as autophagy. Autophagy is the process by which cells eliminate damaged organelles and proteins via the formation of a double-membraned vesicular engulfing organelle termed the autophagosome. See a comprehensive overview of the signaling molecules involved here. Physiologically, autophagy plays critical roles during organismal development and immune response, by regulating growth and cooperating with the adaptive immune system. In disease states, such as neurodegenerative conditions, inefficient autophagic activity leads to the accumulation of abnormal proteins and formation of intracellular aggregates. Despite its clear links to disease and its profound effects on healthspan and lifespan, there are no selective and efficacious autophagy-inducing medicines available.

We caught up with Head of Biology  John K. Blackwood PhD and Jakub Stefaniak PhD, at Samsara Therapeutics to learn how the team are working to discover new mechanisms which can restore autophagy, as well as new drugs which use these mechanisms for the treatment of genetically defined diseases.

Building An Unbiased Data-Driven Drug Discovery Engine

To identify molecules that influence autophagy, the team couple an unbiased screening approach with techniques and instruments, including Jess, to enable deeper understanding of the mechanism and target. ‘Over the past two years the Simple Western™ system continues to be absolutely instrumental in our ability to understand autophagy’s role in disease and beyond. With a vast number of proteins and pathways spanning the length of a cell and responsible for coordinating the cellular recycling process, there was a clear demand for a high throughput immunoblotting method that would allow us to automate traditional western blotting and identify stimulators in diseases where this process is inhibited’ says Dr John K. Blackwood.

Initial Hurdles Transitioning to Automation

It wasn’t all plain sailing at first. ‘To begin with, learning how to incorporate Jess came with some challenges. With the ability to run 25 samples at once it was important to know how to set up the microplate to load our samples and reagents. However, the support and supervision from the Bio-Techne team helped to overcome these initial teething problems and after 3 or 4 runs its use became intuitive’ explained Jakub. Now the team can confidently complete 3 runs with Jess in a single day, condensing months of work to less than a week. Alongside building up their expertise using the instrument, the team have also invested in electronic pipettes to accelerate their rate of data acquisition even further.

New Standards for Data Reproducibility

In addition to automation, Jess enables easy quantification compared to a traditional western blot. ‘Jess allows us to immediately identify potential autophagy biomarkers as they are upregulated or downregulated in response to treatment. This allows us to use more advanced statistical techniques such as principal component analysis and hierarchal clustering to recognize key correlations in our research with ease’ elucidates Blackwood.

Moving forward, Samsara plan to continue to utilise the Jess to develop and leverage their proprietary knowledge alongside transcriptomic and metabolic analysis and build on their characterisation map of autophagy.