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Gene Therapy Validated In Human SCA3 Stem Cells

Written by Dr. Marija Cvetanovic Edited by Dr. Sriram Jayabal

Research group in Michigan report the creation of the first NIH-approved human cell model that mirrors SCA3 disease features – cellular defects that, after gene therapy, show improvement

Spinocerebellar ataxia type 3 (SCA3) is a dominantly-inherited, late onset genetic disease that affects multiple brain regions. Affected individuals suffer from several symptoms, with impaired movement coordination being the most debilitating. SCA3 is caused by a mutation in the Ataxin-3 (ATXN3) gene. In unaffected individuals, ATXN3 typically has anywhere from 12 to 44 repeats of the genetic code “CAG;” however, in some people’s genetic code, the number of CAG repeats can become abnormally high. If this “repeat expansion” mutation causes the ATXN3 gene to have more than 56 CAG repeats, the person develops SCA3. Cells use repeating CAG sequences in their genome to make proteins with long tracts of the amino acid glutamine. In SCA3 cells, these “polyglutamine” (polyQ) tracts are abnormally long in the ATXN3 protein, which makes the protein more prone to form clumps (or “aggregates”) in the cell. The presence of these protein clumps in the cells of the brain is one of the hallmarks of SCA3.

Despite knowing the genetic cause of SCA3, it is still not known how this mutation affects cells on the molecular level. Having said that, several cellular and animal models have been developed in the past two decades to help study these underlying mechanisms. SCA3 models have not only helped to  our increased understanding of the disease’s progression at all levels – molecular, cellular, tissue, and behavioral – but also helped move us closer to therapeutic interventions. For instance, recent studies using SCA3 mouse models have established that targeting ATXN3 with a form of gene therapy known as antisense oligonucleotide (ASO) treatment could very well be an effective strategy for improving the lives of patients. ATXN3-targeting ASOs cause the cells of the brain to produce less of the mutant ATXN3 protein and, when given to SCA3 mice, improved their motor function. These results strongly support the potential use of ASOs to treat SCA3. Still, it is important to see if this finding can be repeated in human neurons (a step that is needed to bring us closer to ASO clinical trials).

Image of a research scientist hard at work in the lab. Image courtesy of pxfuel.

Previous experience from unsuccessful clinical trials highlight the importance of determining the similarities and differences between humans and mice when it comes to disease. For instance, the SCA3 mutation does not naturally occur in mice; therefore, modeling SCA3 with mice usually requires additional genetic manipulation, which could lead to unexpected effects that we do not typically see in patients. In addition, we may miss important determinants of SCA3 pathology due to the inherent differences between humans and mice. For example, proteins that help contribute to SCA3 in human patients may simply not be present in mouse neurons (and vice versa). Because of such species differences, the therapeutic interventions that are effective in mice are not always as effective in humans.

SCA3 human neurons can help bridge the gap between rodent models and human patients, acting as a clinically relevant tool for looking into disease mechanisms and testing new therapies. Because we cannot remove a portion of an SCA3 patient’s brain to study the disease, these neurons must be created in a lab. Human neurons can be generated from induced pluripotent stem cells (iPSCs) or from human embryonic stem cells (hESCs). Induced pluripotent stem cells (iPSCs) are made from adult cells (usually blood or skin cells) that are reprogrammed to return to an embryo-like form (known as the “pluripotent” state). Just like during normal development, iPSCs can create many different types of cells, including neurons. One problem with this approach is that the process of reprograming can potentially change these cells in way that could affect how the disease presents. To avoid this issue, researchers can also create human neurons from human embryonic stem cells (hESCs), which are derived from embryos and are, therefore, naturally pluripotent. Because hESCs do not require reprograming, they are more likely to accurately model disease. However, they are more difficult to obtain and work with. The researchers in this study, led by Lauren Moore in Dr. Hank Paulson’s lab at the University of Michigan, used hESCs to generate the first ever National Institutes of Health (NIH)-approved SCA3 model using human cells.

The team obtained the SCA3 embryonic stem cells from embryos that were originally created for in-vitro fertilization: when the embryos tested positive for the SCA3-causing mutation, the participating families elected to donate them to the University of Michigan instead of implanting them. The researchers found that even before being differentiated into neurons, the SCA3 embryonic stem cells reproduced many hallmarks of SCA3 – including the formation of ATXN3 protein aggregates. And, after forming human SCA3 neurons from these cells, the authors found the same thing. Thus, this is a cell line that advances our ability to model human SCA3, and provides researchers with an essential tool we can use to better understand SCA3 pathology in human neurons. For example, we still do not fully understand whether mutant ATXN3 protein aggregates are bad for neurons. One of the reasons for our lack of understanding in this area is because aggregates do not form easily in many of the SCA3 cell lines created thus far – including human SCA3 iPSCs. In fact, in order for aggregates to form in cells, scientists often need to perform additional chemical or genetic manipulations. Because these procedures have the potential to change the cells in many different ways, using them makes it much harder to interpret any observed results. Thus, this new SCA3 cell line – in which aggregates are easily formed – is likely to significantly increase our understanding of whether ATXN3 aggregates are good or bad for neurons.

That said, the most striking result reported by the researchers here is the validation of a promising SCA3 treatment: gene therapy with ASOs. As mentioned above, ASOs were previously found to be effective in reducing disease symptoms in mouse models of SCA3. In this study, the researchers treat their new human neuron model with ATXN3-targeting ASOs. They found that ASO treatment reduced ATXN3 levels and, most importantly, improved disease-associated defects in the human SCA3 neurons. This represents a significant validation of the power of gene therapy using ATXN3-targeting ASOs, and will further advance its potential to become a successful treatment for SCA3.

In summary, this research team has developed a first-of-its-kind, NIH-approved SCA3 disease model from human ESCs. This model is bound to serve as an important tool for studying the mechanisms of SCA3 and for testing the effectiveness of potential therapeutic approaches – a feature they have already taken advantage of by validating the effectiveness of an ASO treatment strategy for SCA3.

Key Terms

Pluripotency: Ability for one cell to give rise to many different cell types.

Translation: The creation of proteins in cells, where the cell “reads” instruction copied over from DNA into RNA in order to make the final protein. To learn more about translation, read our Snapshot on What is RNA?

Conflict of Interest Statement

The authors and editor declare no conflict of interest.

Four authors on the original paper (L. Moore, D. Bushart,H. McLoughlin, and M. Do Carmo Costa) are contributors to SCAsource. None of these authors had any contribution to the writing or editing of this summary.

Citation of Article Reviewed

Moore, L. R., Keller, L., Bushart, D. D., Delatorre, R. G., Li, D., McLoughlin, H. S., do Carmo Costa, M., Shakkottai, V. G., Smith, G. D., & Paulson, H. L. (2019). Antisense oligonucleotide therapy rescues aggresome formation in a novel spinocerebellar ataxia type 3 human embryonic stem cell line. Stem cell research, 39, 101504.

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