Interaction of Ataxin-1 and DNA repair proteins contributes to SCA1 disease onset and progression

Written by Dr. By Marija Cvetanovic Edited by Dr. Larissa Nitschke

Suart et al. show that Ataxin-1 interacts with an important DNA repair protein Ataxia telangiectasia mutated (ATM), and that reduction of ATM improves motor phenotype in the fruit fly model of SCA1, indicating DNA repair as an important modifier of SCA1 disease progression.

Each day, due to a combination of wear and tear from the normal processes in the cells, and environmental factors, such as irradiation, DNA in each of our cells can accumulate from 10,000 to 1,000,000 damages. If damaged DNA is left unrepaired, this can lead to loss of cell function, cell death, or a mutation that may facilitate the formation of tumors. To avoid these negative outcomes, cells take care of damaged DNA employing DNA damage response/repair proteins. Ataxia-telangiectasia mutated (ATM) protein is a critical part of DNA repair as it can recognize sites of DNA damage. It also helps recruit other proteins that repair DNA damage.

Mutations in the ATM gene cause autosomal recessive ataxia called Ataxia telangiectasia (AT). AT is characterized by the onset of ataxia in early childhood, prominent blood vessels (telangiectasia), immune deficiency, an increased rate of cancer, and features of early ageing.

An artist's drawing of four strands of DNA
DNA repair may be an important modifier of SCA1 disease progression. Photo used under license by Anusorn Nakdee/

Expansion of CAG repeats in the Ataxin-1 gene causes dominantly inherited Spinocerebellar Ataxia Type 1 (SCA1). A feature of SCA1 is that a greater number of repeats correlates to an earlier age of onset of symptoms and worse disease progression. The connection of DNA repair pathways and SCA1 was brought into focus in 2016 by a study by Bettencourt and colleagues. As longer CAG repeat tracts association with earlier ages at onset do not account for all of the difference in the age of onset authors searched for additional genetic modifying factors in a cohort of approximately 1000 patients with SCAs. They showed that DNA repair pathways significantly associate with the age at onset in SCAs, suggesting that genes with roles in the DNA damage response could provide new therapeutic targets (and hence therapeutics) in SCAs.

In this study, Suart et al. identify ATM as one such gene. Using irradiation and oxidizing agent to damage DNA and using imaging to follow ataxin-1 movement, authors first show that ataxin-1 is recruited to the site of DNA damage in cultured cells. They also demonstrate that SCA1 mutation slows down but does not prevent ataxin-1 recruitment to the sites of DNA damage.

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Newly identified mutations in SCA19/22 and their dysfunctions

Written by Sophia Leung Edited by Dr. Marija Cvetanovic

While the mutant proteins in SCA19/22 lose part of their innate functions and properties, they also disrupt the key functions of the normal healthy protein.

The underlying mechanism of the hereditary property of SCA19/22 is elusive. In this study, the researchers investigated the molecular properties of four different mutations found in patients with SCA19/22. They looked at how these mutant proteins affect the normal protein if they are both present in the cell. They found that the mutant proteins are not only non-functional (do not work properly), but that in their presence, the normal protein’s function is also diminished. Furthermore, while the production and proper localization of these mutant proteins are found to be defective, they also bring the same decline to the normal protein. This adds to their disease-causing properties. This study is significant in that it offers a molecular investigation into mutant proteins associated with SCA19/22 that was previously lacking. It also provides evidence that may explain the hereditary property of the disease.

A number of mutations in the gene KCND3 has been associated with SCA19/22. The gene makes the voltage‐gated potassium ion (K+) channel subunit KV4.3. In general terms, the gene makes a protein that functions to allow potassium ions to pass through the membrane of nerve cells. Similar to how a flute has many holes to allow air to pass through when played to make a specific note, a nerve cell has different kinds of channels to allow ions to pass through their membrane to orchestrate normal functioning. One could imagine the disruption to any channels, a partial obstruction or a total blockage, could perturb the overall output of the cell.

flute resting on a music stand
Similar to how a flute has many holes to allow air to pass through when played to make a specific note, a nerve cell has different kinds of channels to allow ions to pass through to orchestrate normal functioning. (Photo by Rajesh Kavasseri / Unsplash)

In this study, the researchers found that the normal KV4.3 channel protein detectably allows potassium ions to pass through. But little to no ions can pass through the SCA19/22 mutant KV4.3 channels. Even under the assistance of a “helper” protein, which normally enhances the function of this channel, only one of the mutant channel proteins shows improvement. This indicates that the SCA19/22 causing mutations result in a reduced function of mutated KV4.3 channels.

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Evaluating the long-term safety of lentiviral gene therapy in SCA3 mice

Written by Dr. Ambika Tewari Edited by Dr. Hayley McLoughlin

Lentiviral expression of an shRNA against ataxin-3 was well-tolerated and produced no measurable adverse effects in wild-type mice.

Evaluating the safety profile is a necessary and crucial step in qualifying a therapy for use in patients. Gene therapy is an experimental technique that has demonstrated tremendous progress in the treatment or reversal of a disease, specifically monogenic disorders. Carefully investigating the safety and tolerance of gene therapy is important to gauge its suitability for clinical trials. Gene therapy tools can be used in different ways to achieve the same therapeutic effect: the faulty gene can be replaced with a healthy copy, the mutated gene can be repaired, or the mutant copy of the gene can be silenced. You can learn more about gene therapy in this pat SCAsource Snapshot.

Spinocerebellar ataxia type 3 (SCA3) or Machado-Joseph disease (MJD) causes progressive loss of neurons in the spinal cord, and several regions of the brain. This includes the cerebellum, brainstem, striatum and substantia nigra. These neurons have crucial functions. Without these neurons, patients experience motor incoordination, loss of balance, and in severe cases, premature death. While great progress continues to be made in understanding how a mutation in a single gene, Ataxin-3, causes the symptoms of SCA3, there is still no treatment to stop the disease progression. As a monogenic disorder, SCA3, like other Spinocerebellar ataxias (SCA), is a promising candidate for gene therapy. While there are no approved gene therapies for SCA yet, there any several research labs and companies working towards achieving this goal.

An artist's drawing of scientists standing infront of a giant piece of DNA and drugs
This is truly an exciting time for gene therapy, but it is also important to keep the safety of patients a top priority. Photo used under license by Visual Generation/

The researchers in this study have been working on gene therapy for SCA3 since 2008. They have researched how gene therapy could offer protection against further decline, in several cell and mouse models of SCA3. They used an approach where they decreased the levels of the mutant Ataxin-3 gene while leaving the normal Ataxin-3 gene intact. This is known as allele-specific targeting. They demonstrated that using this technique, they could significantly reduce the behavioral and neuropathological changes that occur in SCA3 mice. Mice treated with the gene therapy showed improvements in their balance and motor coordination. 

Gene therapy in its most basic form involves two components, the gene that will replace or remove the diseased gene and a vector that will transport this new gene to its site of action. The most commonly used vectors today are adeno-associated virus (AAVs) followed by retrovirus. These viruses have been specifically engineered to deliver their passenger to the specified location. While both vectors have been through several years of preclinical and clinical testing for numerous gene therapy candidates, there are questions that remain regarding their safety. (1) Does the gene therapy product continue to be expressed in the targeted area long-term; (2) If there is long-term expression does it cause any adverse measurable effects to the targeted area; (3) Does the long-term expression affect the normal functioning of the targeted cells/organ.

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Regulating ataxin-1 expression as a therapeutic avenue for SCA1

Written by Dr. Hannah Shorrock   Edited by Dr. Hayley McLoughlin

Nitschke and colleagues identify a microRNA that regulates ataxin-1 levels and rescues motor deficits in a mouse model of SCA1

What if you could use systems already in place in the cell to regulate levels of toxic proteins in disease? This is the approach that Nitschke and colleagues took to identify the cellular pathways that regulate ataxin-1 levels. Through this strategy, the group found a microRNA, a small single-stranded RNA, called miR760, that regulates levels of ataxin-1 by directly binding to its mRNA and inhibiting expression. By increasing levels of miR760 in a mouse model of SCA1, ataxin-1 protein levels decreased and motor function improved. This approach has the potential to identify possible therapies for SCA1. It may also help identify disease-causing mutations in ataxia patients with unknown genetic causes.

Spinocerebellar Ataxia type 1 (SCA1) is an autosomal dominant disease characterized by a loss of coordination and balance. SCA1 is caused by a CAG repeat expansion in the ATXN1 gene. This results in the ataxin-1 protein containing an expanded polyglutamine tract. With the expanded polyglutamine tract, ataxin-1 is toxic to cells in the brain and leads to dysfunction and death of neurons in the cerebellum and brainstem.

As with all protein-coding genes, surrounding the protein coding region of ATXN1 gene are the 5’ (before the coding sequence) and 3’ (after the coding sequence) untranslated regions (UTRs). These regions are not translated into the final ataxin-1 protein product but are important for the regulation of this process. Important regulation factors called enhancers and repressors of translation located in 5’ and 3’ UTRs. ATXN1 has a long 5’ UTR. Genes that require fine regulation, such as growth factors, are often found to have long 5’ UTRs: the longer a 5’ UTR, the more opportunity for regulation of gene expression. The group, therefore, tested the hypothesis that the 5’ UTR is involved in regulating the expression of ataxin-1.

In their initial studies, Nitschke and colleagues identified that the ATXN1 5’UTR is capable of reducing both protein and RNA levels when placed in front of (5’ to) a reporter coding sequence. One common mechanism through which this regulation of gene expression could be occurring is the binding of microRNAs, or miRNAs, to the ATXN1 5’UTR. miRNAs are short single-stranded RNAs that form base pairs with a specific sequence to which the miRNA has a complementary sequence; this leads to regulation of expression of the mRNA to which the miRNA is bound.

3d illustration of single-strand ribonucleic acid
Artist drawing of single-stranded RNA. Photo used under license by nobeastsofierce/

Using an online microRNA target prediction database called miRDB, the group identified two microRNAs that could be responsible for these changes in gene expression through binding to the ATXN1 5’ UTR. By increasing the expression of one of these microRNAs, called miR760, ataxin-1 protein levels were reduced in cell culture. Conversely, using a miR760 inhibitor so that the miRNA could not perform its normal functions led to increased levels of ataxin-1. Together this shows that miR760 negatively regulates ataxin-1 expression.

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Identifying FDA-approved molecules to treat SCA6

Written by Dr Hannah Shorrock Edited by Dr. Larissa Nitschke

Pastor and colleagues identify FDA-approved small molecules that selectively reduce the toxic polyglutamine-expanded protein in SCA6.

Selectively targeting disease-causing genes without disrupting cellular functions is essential for successful therapy development. In spinocerebellar ataxia type 6 (SCA6), achieving this selectivity is particularly complicated as the disease-causing gene produces two proteins that contain an expanded polyglutamine tract. In this study, Pastor and colleagues identified several Food and Drug Administration (FDA) approved small molecules that selectively reduce the levels of one of these polyglutamine-containing proteins without affecting the levels of the other protein, which is essential for normal brain function. By using drugs already approved by the United States Food and Drug Administration to treat other diseases, referred to as FDA-approved drugs, the team hopes to reduce the time frame for pre-clinical therapy development.

SCA6 is an autosomal dominant ataxia that causes progressive impairment of movement and coordination. This is due to the dysfunction and death of brain cells, including Purkinje neurons in the cerebellum. SCA6 is caused by a CAG repeat expansion in the CACNA1A gene. CACNA1A encodes two proteins: the a1A subunit, the main pore-forming subunit of the P/Q type voltage-gated calcium ion channel, as well as a transcription factor named a1ACT.

The a1A subunit is essential for life. Its function is less affected by the presence of the expanded polyglutamine tract than that of a1ACT. The transcription factor, a1ACT, controls the expression of various genes involved in the development of Purkinje cells. Expressing a1ACT protein containing an expanded polyglutamine tract in mice causes cerebellar atrophy and ataxia. While reducing levels of the a1A subunit may have little effect on SCA6 disease but impact normal brain cell function, reducing levels of a1ACT may improve disease in SCA6. Therefore, Pastor and colleagues decided to test the hypothesis that selectively reducing levels of the a1ACT protein without affecting levels of the a1A protein may be a viable therapeutic approach for SCA6.

Colorful pile of medicines in blister packs which color are White, Yellow, Black and Pink pills.
By using drugs already approved by the FDA, the team hopes to reduce the time frame for pre-clinical therapy development. Photo used under license by Wanchana Phuangwan/
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