Clearing aggregated ataxin-2 protein as a therapeutic avenue for SCA2

Written by Dr. Vitaliy Bondar Edited by Dr. Hayley McLoughlin

New research suggests that mutant ataxin-2 protein overwhelms cells in SCA2, leading to decreased autophagy and clearance of damaged proteins.

Many comparisons can be made between cells and human beings. Just like humans, cells can accumulate junk and waste at certain times and this clutter overtime becomes problematic and even toxic. This is precisely what Jonathan Henry Wardman and colleagues from the University of Copenhagen decided to investigate on a cellular level. They asked whether the lack of appropriate clearance of faulty disease proteins effect cellular survival and wellbeing.

The researchers chose to study cells derived from a patient that has Spinocerebellar ataxia type 2 (SCA2). The cause of SCA2 is CAG repeat expansion in the ATAXIN-2 gene, which encodes polyglutamine amino acid chain in an RNA-binding protein, ataxin-2. The faulty polyQ expanded ATXN2 protein is found to aggregate inside the cell and overtime can affect its survival. Accumulation of aggregated protein products derived from mutated genes is a hallmark of many types of spinocerebellar ataxias as well as other forms of neurodegenerative disorders such as Parkinson’s disease.

It is unclear how protein aggregation impacts cellular survival. However, multiple cellular defects have been correlated with ataxin-2 aggregation. For instance, mitochondria which generates energy for a cell, has been reported to abnormally function in SCA2 cellular models. Additionally, a cellular clearance mechanism, called autophagy, which is responsible for clearing faulty cellular compartments and certain broken proteins is shown to be less effective in various SCA2 models. These mechanisms the authors decided to investigate in their recently published research article.

scientist using microscope
New research using SCA2 cells sheds light on what causes disease symptoms to occur. Photo by Chokniti Khongchum on

The scientists first identified evidence of SCA2 cellular dysfunction through detection of significant elevation of caspase-9 and caspase-8 levels. These are protein which indicate cellular stress and death. The authors hypothesized that such cellular dysfunction may arise from accumulation of faulty ataxin-2. In order to test this hypothesis, they decided to systematically block two cellular pathways that process defective proteins: proteostasis and autophagy.

Continue reading “Clearing aggregated ataxin-2 protein as a therapeutic avenue for SCA2”

ASO treatment to lower ataxin-1 levels doesn’t cause unwanted side effects in a SCA1 mouse model

Written by Dr. Ronald Buijsen Edited by Dr. Larissa Nitschke

O’Callaghan and colleagues show that novel therapeutic approaches to reduce the disease-causing protein in SCA1 do not increase the risk of developing cancer or Alzheimer’s disease in SCA1 mice.

People affected with Spinocerebellar Ataxia type 1 or SCA1 carry an expansion of a repetitive stretch of DNA in the ATXN1 gene. The expanded ATXN1 gene encodes an expanded ataxin-1 protein, which accumulates and causes toxicity in the brain. This causes motor coordination problems and premature lethality. So far, there is no treatment that slows, stops, or reverses SCA1 disease progression.

Still, several preclinical studies demonstrated that reducing ataxin-1 protein levels can improve the motor coordination deficits in SCA1 mouse models. One strategy to reduce ataxin-1 levels is the use of antisense oligonucleotides (ASO). These ASO treatments specifically cleave Atxn1 mRNA and lower ataxin-1 protein levels.

This study, published by the group of Dr. Harry Orr in 2018, showed that injection of ASOs into the brain of SCA1 mice improves motor deficits, prolonged survival, and reversed neurochemical abnormalities. However, lowering ataxin-1 protein levels might lead to altered expression of other proteins in the brain. This could impact the safety of this treatment strategy. Therefore, this follow-up study investigated whether lowering of ataxin-1 protein levels results in unwanted effects.

a brown laboratory mouse sits in a researcher's gloved hand
ASO research in SCA1 is promising. But before moving forward, more safety testing had to be done in SCA1 mouse models. Image courtesy of Rama on Wikimedia.
Continue reading “ASO treatment to lower ataxin-1 levels doesn’t cause unwanted side effects in a SCA1 mouse model”

2 minuti di Scienza: Cosa sono I nucleotidi antisenso?

I nucleotidi anti-senso (anche noti come ASOs o AON, dall’inglese Antisense oligonucleotides) sono piccole molecole che possono essere usate per prevenire o alterare la produzione di proteine. Le proteine sono la forza lavoro della cellula, e dirigono la maggior parte dei processi cellulari. Le proteine sono prodotte in due fasi: nella prima un gene che codifica per una proteina viene convertito in una molecola che contiene specifiche istruzioni, l’RNA messaggero (mRNA). L’ mRNA trasferisce l’informazione contenuta nei geni al compartimento che assembla le proteine. Qui, l’mRNA è infine trasformato in proteina. Gli ASOs sono corte sequenze di DNA a singolo filamento, complementari alla sequenza di uno specifico mRNA. In base a diversi tipi di modifiche chimiche della loro sequenza, gli ASOs possono determinare due tipi di effetti sull’ mRNA complementare. Alcune modifiche fanno si che gli ASO distruggano l’mRNA e, di conseguenza, causano la perdita della proteina corrispondente. Altre modifiche, invece, permettono agli ASO di mascherare certi tratti dell’mRNA bersaglio, causando la produzione di una versione alterata della proteina.

Come funzionano gli ASO nel corpo umano. Autore della figura Larissa Nitschke, creato con BioRender.

La maggior parte delle Atassie spinocerebellari (dall’inglese Spinocerebellar Ataxias, SCAs) sono causate dall’accumulazione di una proteina tossica in una specifica regione del cervello. Per questo motivo, il principale obiettivo del trattamento delle SCAs con gli ASOs è inibire la produzione della proteina tossica. Un esempio di questa applicazione degli ASO è il lavoro del Prof. Harry Orr all’ Università del Minnesota. Il suo gruppo di ricerca studia l’Atassia spinocerebellare di tipo 1 (SCA1), causata dall’accumulo tossico della proteina Ataxina-1. Iniezioni di ASOs in modelli animali di SCA1 riducono i livelli di Ataxina-1 e migliorano l’incoordinazione motoria tipica della SCA1. Un altro modo di usare gli ASOs per il trattamento delle SCAs è la modifica dell’informazione trasmessa dall’mRNA per produrre una versione alterata della proteina. Questo approccio è stato testato nel caso della Atassia spinocerebellare di tipo 3 (SCA3), nella quale un’espansione nel gene Atxn3 rende la proteina Ataxina 3 tossica. Il gruppo del Dr. van Roon-Mom, in Olanda, per esempio, ha usato gli ASOs per rimuovere esclusivamente la porzione espansa della proteina Atxn3, lasciando intatta il resto della struttura proteica e la sua funzione.

Entrambi gli studi, così come altri studi portati avanti per altre SCAs, hanno evidenziato il potenziale uso degli ASOs come strumenti terapeutici per le SCAs. Mentre la ricerca sugli ASOs per le SCAs è per lo più nella fase preclinica, il trattamento con gli ASO per altre malattie, come la Distrofia Muscolare di Duchenne e l’atrofia muscolare spinale, è stato già approvato dall’ente statunitense Food and Drug Administration. Ulteriori studi clinici saranno necessari per misurare il beneficio terapeutico degli ASOs in pazienti di SCAs.

Per saperne di più sugli oligonucleotidi antisenso, leggi questo articolo alla pagina  HDBuzz sugli ASOs in via di sviluppo per la malattia di Huntington.

“2 minuti di Scienza” scritto da Dr. Larissa Nitschke, revisionato da Dr. Hayley McLoughli, tradotto in italiano da Dr. Antonia De Maio. Pubblicato per la prima volta il 31 Maggio 2019.

A new molecule identified that controls cerebellar communication

Written by Dr. Ambika Tewari Edited by Dr. Sriram Jayabal

Targeting phosphatases in the cerebellum can correct miscommunication in multiple models of ataxia.

The cerebellum is essential for motor coordination and consists of the coordinated activity of different types of cells. Purkinje cells are one of the most fascinating cell types in the cerebellum. They have an elaborate network of branches called dendrites, where a neuron receives communication from other neurons. It is one of the most complex branching systems seen across all neurons in the entire brain. Each one of these branches has many points of contact with other branches called axons. Each axon is part of a neuronal structure that allow communication between neurons. These axons are from different cell types and allow information to be transferred to Purkinje cells.

Colourful illustration of a human brain
Targeting phosphatases in the brain could improve communication between neurons, reducing ataxia symptoms.

Due to this branching complexity, Purkinje cells receive many messages or inputs. This represents different pieces of sensory information to ensure that movements are precisely timed. Purkinje cells must integrate and process this information. This produces motor behaviors like walking, writing, playing a musical instrument, and many more. Any alteration to the processing of this information will result in cerebellum dysfunction; in fact, Purkinje cells have gained attention because they undergo progressive deterioration in most ataxias. 

Neurons, including Purkinje cells, communicate with other neurons using electrical signals known as action potentials or spikes. Firing rate, defined as the number of spikes within a defined period of time, is thought to be an important feature of this communication, which is critical for coordinating muscle movements. Therefore, a lower firing rate in Purkinje cells would signal a faulty communication between Purkinje cells and their targets. This has devastating consequences as seen in many ataxias.

For instance, in an earlier study, a group of authors found that the firing rate of Purkinje cells was decreased in mouse models of three different Spinocerebellar ataxias (SCAs): SCA1, SCA2, and SCA5. They further explored whether there was a common reason underlying the decreased firing rate. They found that a protein named Missing in Metastasis (MTSS1), was important for Purkinje cells to effectively communicate with each other. Mice engineered to have no MTSS1 protein had a decreased firing rate and difficulty walking and maintaining their balance.

In every cell in the body, including brain cells, there are numerous proteins that perform different functions. The concerted effort of all are needed for the cell to perform its intended duty. Some of these proteins are maintained in the cell in an inactive form and are activated when they are required in the cell and inhibited when they are not. This highly regulated system aims to maintain precise levels of proteins in each cell, while simultaneously conserving energy. Each cell has many ways of activating/inactivating a protein. A specialized group of proteins known as kinases and phosphatases, adds and removes phosphate groups to and from proteins respectively, thereby altering their active/inactive forms which then changes their interactions with other proteins. MTSS1 is one such protein that inhibits the activity of a group of kinases known as Src family of non-receptor tyrosine kinases (SFKs).

Continue reading “A new molecule identified that controls cerebellar communication”

Spotlight: The Neuro-D lab Leiden

Principal Investigator: Dr. Willeke van Roon-Mom

Location: Leiden University Medical Centre, Leiden, The Netherlands

Year Founded: 1995

What disease areas do you research?

What models and techniques do you use?

A group photo of members of the Neuro-D lab Leiden standing outside on a patio.
This is a group picture taken during our brainstorm day last June. From left to right: Boyd Kenkhuis, Elena Daoutsali, Tom Metz, Ronald Buijsen, Willeke van Roon-Mom (PI), David Parfitt, Hannah Bakels, Barry Pepers, Linda van der Graaf and Elsa Kuijper. Image courtesy of Ronald Buijsen.

Research Focus

What is your research about?

The Neuro-D research group studies how diseases develop and progress at the molecular level in several neurodegenerative diseases. They focus on diseases that have protein aggregation, where the disease proteins clump up into bundles in the brain and don’t work correctly.

We focus strongly on translational research, meaning we try to bridge the gap between research happening in the laboratory to what is happening in medical clinics. To do this we use more “traditional” research models like animal and cell models. But we also use donated patient tissues and induced pluripotent stem cell (iPSC) models, which is closer to what is seen in medical clinics.

Our aim is to unravel what is going wrong in these diseases, then discover and test potential novel drug targets and therapies.

One thing we are doing to work towards this goal is identifying biomarkers to measure how diseases progress over time. To do this, we use sequencing technology and other techniques to look at new and past data from patients.

Why do you do this research?

So far there are no therapies to stop the progression of ataxia. If we can understand what is happening in diseases in individual cells, we can develop therapies that can halt or maybe even reverse disease progression.

Identifying biomarkers is also important, because it will help us figure out the best time to treat patients when we eventually have a therapy to test.

Stylized logo for the Dutch Center for RNA Therapeutics
The Neuro-D lab Leiden is part of the Dutch Center for RNA Therapeutics, which focuses on RNA therapies like antisense oligonucleotides. Logo designed by Justus Kuijer (VormMorgen), as 29 year old patient with Duchenne muscular dystrophy.

Are you recruiting human participants for research?

Yes, we are! We are looking for participants for a SCA1 natural history study and biomarker study. More information can be found here. Please note that information about this study is only available in Dutch.

Fun Fact

All our fridges and freezers have funny names like walrus, seal, snow grouse and snowflake.

For More Information, check out the Neuro-D lab Leiden website!

Written by Dr. Ronald Buijsen, Edited by Celeste Suart