RARE DISEASE
Canavan disease
Canavan disease
Canavan disease
Synonyms: ACY2 deficiency, Aminoacylase 2 deficiency, Aspartoacylase deficiency, Spongy degeneration of the brain
Synonyms: ACY2 deficiency, Aminoacylase 2 deficiency, Aspartoacylase deficiency, Spongy degeneration of the brain
Synonyms: ACY2 deficiency, Aminoacylase 2 deficiency, Aspartoacylase deficiency, Spongy degeneration of the brain
Drug discovery
4
drugs
With orphan designations
Overview
Canavan disease is a rare autosomal recessive leukodystrophy caused by ASPA gene mutations, leading to deficient aspartoacylase enzyme activity and toxic accumulation of N-acetyl-aspartate (NAA) in the brain. This disrupts myelination, causing progressive neurodegeneration. Clinical features include macrocephaly, hypotonia, developmental regression, seizures, and early death in severe infantile forms. Milder juvenile variants present with nonspecific motor/speech delays [1][9][16].
Therapies
Symptomatic management: Anticonvulsants, nutritional support, and respiratory care [14][16]
Investigational gene therapy: Intracerebroventricular rAAV-Olig001-ASPA shows reduced NAA levels and improved myelination in early trials [3][5][17]
Enzyme replacement and acetate supplementation under preclinical study [11][14]
Categories: rare genetic diseases, rare inborn errors of metabolism, rare neurological diseases
Research Papers
136 drug discovery papers about Canavan disease, with 2 first-in-class and 1 next-in-class emerging drug candidates forecasted to outperform the average preclinical success rate. Recent publications:
136 drug discovery papers about Canavan disease, with 2 first-in-class and 1 next-in-class emerging drug candidates forecasted to outperform the average preclinical success rate. Recent publications:
categories:
Small molecules
small molecules
2026-02-12 | Discovery of N-Acetyltransferase 8-Like (NAT8L) inhibitors based on a N-Acylated (Piperidin-3-ylmethyl)-1,2,4-Oxadiazole Scaffold.
Canavan disease (CD) is an autosomal recessive genetic disorder caused by mutations in the ASPA gene, which encodes the enzyme aspartoacylase. These mutations lead to a deficient enzymatic activity and increased concentrations of its substrate, N-acetylaspartate (NAA), in the brain and other tissues. Aspartate N-acetyltransferase, encoded by the N-acetyltransferase 8-like (NAT8L) gene, catalyzes the biosynthesis of NAA from aspartate and acetyl-CoA. Therefore, inhibition of NAT8L has been implicated as a promising therapeutic strategy for CD by normalizing NAA levels in the brain. Our high throughput screening campaign followed by a rigorous hit validation process identified 2-(2-fluorophenoxy)-1-(3-((3-(thiophen-3-yl)-1,2,4-oxadiazol-5-yl)methyl)piperidin-1-yl)ethan-1-one (4a) as a low micromolar, noncarboxylic acid inhibitor of NAT8L. Subsequent structural optimization led to the discovery of two submicromolar NAT8L inhibitors. Although these inhibitors displayed high clearance in liver microsomes, the new scaffold, devoid of a carboxylic acid moiety, could potentially lead to potent and brain-penetrant NAT8L inhibitors through further molecular refinement.
2024-11-26 | N-acetylaspartate mitigates pro-inflammatory responses in microglial cells by intersecting lipid metabolism and acetylation processes.
Microglia play a crucial role in brain development and repair by facilitating processes such as synaptic pruning and debris clearance. They can be activated in response to various stimuli, leading to either pro-inflammatory or anti-inflammatory responses associated with specific metabolic alterations. The imbalances between microglia activation states contribute to chronic neuroinflammation, a hallmark of neurodegenerative diseases. N-acetylaspartate (NAA) is a brain metabolite predominantly produced by neurons and is crucial for central nervous system health. Alterations in NAA metabolism are observed in disorders such as Multiple Sclerosis and Canavan disease. While NAA's role in oligodendrocytes and astrocytes has been investigated, its impact on microglial function remains less understood. The murine BV2 microglial cell line and primary microglia were used as experimental models. Cells were treated with exogenous NAA and stimulated with LPS/IFN-γ to reproduce the pro-inflammatory phenomenon. HPLC and immunofluorescence analysis were used to study lipid metabolism following NAA treatment. Automated fluorescence microscopy was used to analyze phagocytic activity. The effects on the pro-inflammatory response were evaluated by analysis of protein/mRNA expression and ChIP assay of typical inflammatory markers. NAA treatment promotes an increase in both lipid synthesis and degradation, and enhances the phagocytic activity of BV2 cells, thus fostering surveillant microglia characteristics. Importantly, NAA decreases the pro-inflammatory state induced by LPS/IFN-γ via the activation of histone deacetylases (HDACs). These findings were validated in primary microglial cells, highlighting the impact on cellular metabolism and inflammatory responses. The study highlighted the role of NAA in reinforcing the oxidative metabolism of surveillant microglial cells and, most importantly, in buffering the inflammatory processes characterizing reactive microglia. These results suggest that the decreased levels of NAA observed in neurodegenerative disorders can contribute to chronic neuroinflammation.
2024-11-14 | Restoring Function in Pediatric Neurodegeneration: The Impact of Radio Electric Asymmetric Conveyor Neuroregenerative Treatment in a Child With Canavan Syndrome.
This case report presents the therapeutic impact of radio electric asymmetric conveyor (REAC) neuroregenerative medicine treatment (REAC RGN-N) in a 10-year-old girl diagnosed with Canavan syndrome, a rare genetic disorder marked by progressive white matter degeneration in the brain. This condition, caused by aspartoacylase deficiency, leads to an accumulation of N-acetyl-L-aspartic acid (NAA), resulting in severe motor impairment, muscle tone abnormalities, and developmental delays. The patient received REAC RGN-N treatment, administered via the REAC - BENE mod 110 (ASMED®, Scandicci, Italy), specifically configured for RGN-N therapy. The protocol consisted of four 6-hour sessions completed over a 24-hour cycle. Following this initial treatment, the patient showed notable improvements in head and trunk control, muscle tone, and swallowing function. Head and trunk control increased from 20 to 30 on the trunk control measurement scale (TCMS), spasticity decreased from a score of 3 to 1 on the Ashworth scale, and her swallowing ability progressed from level 5 to level 7 on the functional oral intake scale (FOIS), allowing her to handle a wider range of food textures independently. These improvements were assessed six months after the end of the first treatment cycle, suggesting a degree of stability in the therapeutic effects observed. Although further studies could support these findings, the rarity of Canavan syndrome presents challenges for conducting additional targeted research. REAC RGN-N treatment is typically repeated in cycles every 4-6 months, depending on clinical severity, to sustain therapeutic benefits. This case report offers initial evidence that REAC RGN-N may provide meaningful, stable improvements for pediatric patients with neurodegenerative conditions, addressing motor and sensory impairments with potential long-term benefits.
2024-08-12 | Identification of potential pharmacological chaperones that selectively stabilize mutated Aspartoacylases in Canavan disease.
Canavan disease is caused by mutations in the ASPA gene, leading to diminished catalytic activity of aspartoacylase in the brain. Clinical missense mutations are found throughout the enzyme structure, with many of these mutated enzymes having not only decreased activity but also compromised stability. High-throughput screening of a small molecule library has identified several compounds that significantly increase the thermal stability of the E285A mutant enzyme, the most predominant clinical mutation in Canavan disease, while having a negligible effect on the native enzyme. Based on the initial successes, some structural analogs of these initial hits were selected for further examination. Glutathione, NAAG and patulin were each confirmed to be competitive inhibitors, indicating the binding of these compounds at the dimer interface or near the active site of the E285A enzyme. The experimental results were theoretically examined with the help of the docking analysis method. The structure activity-guided optimization of these compounds can potentially lead to potential pharmacological chaperones that could alleviate the detrimental effect of ASPA mutations in Canavan patients.
2024-01-28 | Astroglial conditional Slc13a3 knockout is therapeutic in murine Canavan leukodystrophy.
Abstract Canavan disease is a leukodystrophy caused by ASPA mutations that diminish oligodendroglial aspartoacylase activity, and is characterized by markedly elevated brain concentrations of the aspartoacylase substrate N‐acetyl‐ l ‐aspartate (NAA) and by astroglial and intramyelinic vacuolation. Astroglia express NaDC3 (encoded by SLC13A3 ), a sodium‐coupled transporter for NAA and other dicarboxylates. Astroglial conditional Slc13a3 deletion in aspartoacylase‐deficient Canavan disease model mice (“CD mice”) reversed brain NAA elevation and improved motor function. These results demonstrate that astroglial NaDC3 contributes to brain NAA elevation in CD mice, and suggest that suppressing astroglial NaDC3 activity would ameliorate human Canavan disease.
cell therapies
2026-01-24 | Human iPSC-derived neural progenitor cells rescue motor function and brain pathology in symptomatic Canavan disease mice.
Canavan disease (CD) is a severe neurodegenerative disorder caused by aspartoacylase (ASPA) deficiency, leading to N-acetyl-L-aspartic acid (NAA) accumulation and spongy degeneration. While several therapeutic candidates improve outcomes in CD mouse models when delivered before symptom onset, there remains a need for treatments targeting established disease pathology. Here, we demonstrate that transplantation with human induced pluripotent stem cell (iPSC)-derived neural progenitor cells (NPCs) expressing a functional ASPA gene (ASPA iNPCs) can rescue disease manifestations in symptomatic CD (Nur7) mice. When administered at postnatal day 21 (P21), ASPA iNPCs successfully engrafted, differentiated into neural lineage cells, and restored ASPA activity as revealed by reduced NAA level. Transplanted mice showed a significant reduction in brain and cerebrospinal fluid (CSF) NAA levels, decreased vacuolation across multiple brain regions, improved myelination, and enhanced motor function 6-month post-transplantation. Our findings demonstrate that ASPA iNPC transplantation can effectively reverse established CD pathology, suggesting therapeutic potential for treating symptomatic patients.
2023-06-04 | Developing Hypoimmunogenic Human iPSC‐Derived Oligodendrocyte Progenitor Cells as an Off‐The‐Shelf Cell Therapy for Myelin Disorders
Demyelinating disorders are among the most common and debilitating diseases in neurology. Canavan disease (CD) is a lethal demyelinating disease caused by mutation of the aspartoacylase (ASPA) gene, which leads to the accumulation of its substrate N-acetyl-l-aspartate (NAA), and consequently demyelination and vacuolation in the brain. In this study, hypoimmunogenic human induced pluripotent stem cell (iPSC)-derived oligodendrocyte progenitor cells (OPC) are developed from a healthy donor as an "off-the-shelf" cell therapy. Hypoimmunogenic iPSCs are generated through CRISPR/Cas9 editing of the human leukocyte antigen (HLA) molecules in healthy donor-derived iPSCs and differentiated into OPCs. The OPCs are engrafted into the brains of CD (nur7) mice and exhibit widespread distribution in the brain. The engrafted OPCs mature into oligodendrocytes that express the endogenous wildtype ASPA gene. Consequently, the transplanted mice exhibit elevated human ASPA expression and enzymatic activity and reduced NAA level in the brain. The transplanted OPCs are able to rescue major pathological features of CD, including defective myelination, extensive vacuolation, and motor function deficits. Moreover, the hypoimmunogenic OPCs exhibit low immunogenicity both in vitro and in vivo. The hypoimmunogenic OPCs can be used as "off-the-shelf" universal donor cells to treat various CD patients and many other demyelinating disorders, especially autoimmune demyelinating diseases, such as multiple sclerosis.
2014-01-06 | Advances and pitfalls of cell therapy in metabolic leukodystrophies.
Leukodystrophies are a group of disorders characterized by myelin dysfunction, either at the level of myelin formation or maintenance, that affect the central nervous system (CNS) and also in some cases, to a lesser extent, the peripheral nervous system (PNS). Although these genetic-based disorders are generally rare, all together they have a significant impact in the society, with an estimated overall incidence of 1 in 7,663 live births. Currently, there is no cure for leukodystrophies, and the development of effective treatments remains challenging. Not only leukodystrophies generally progress very fast, but also most are multifocal needing the simultaneous targeting at multiple sites. Moreover, as the CNS is affected, the blood-brain barrier (BBB) limits the efficacy of treatment. Recently, interest on cell therapy has increased, and the leukodystrophies for which metabolic correction is needed have become first-choice candidates for cell-based clinical trials. In this review, we present and discuss the available cell transplantation therapies in metabolic leukodystrophies including fucosidosis, X-linked adrenoleukodystrophy, metachromatic leukodystrophy, Canavan disease, and Krabbe's disease. We will discuss the latest advances of cell therapy and its pitfalls in this group of disorders, taking into account, among others, the limitations imposed by reduced cell migration in multifocal conditions, the need to achieve corrective enzyme threshold levels, and the growing awareness that not only myelin but also the associated axonopathy needs to be targeted in some leukodystrophies.
2004-10-01 | Mouse neural progenitor cells differentiate into oligodendrocytes in the brain of a knockout mouse model of Canavan disease
Canavan disease (CD) is an autosomal recessive disorder that leads to spongy degeneration in the white matter of the brain. Aspartoacylase (ASPA) synthesizing cells, oligodendrocytes, are lost in CD. Transplantation of neural progenitor cells (NPCs) offers an interesting therapeutic approach for treating neurodegenerative diseases by replacing the lost cells. Therefore, the NPCs transplantation to the brain of the CD mouse was studied. Injection of mouse NPCs to the striatum and cerebellum of juvenile CD mouse showed numerous BrdU positive cells at 1 month after injection. The same result was also observed in the adult CD mouse brain after 5 weeks of post-transplantation period. The implanted cells differentiated into oligodendrocytes and fibrous astrocytes, as observed using glial cell marker. This is the first report to describe the survival, distribution and differentiation of NPCs within the brain of CD mouse and a first step toward the potential clinical use of cell therapy to treat CD.
2002-05-01 | Global gene and cell replacement strategies via stem cells
The inherent biology of neural stem cells (NSCs) endows them with capabilities that not only circumvent many of the limitations of other gene transfer vehicles, but that enable a variety of novel therapeutic strategies heretofore regarded as beyond the purview of neural transplantation. Most neurodegenerative diseases are characterized not by discrete, focal abnormalities but rather by extensive, multifocal, or even global neuropathology. Such widely disseminated lesions have not conventionally been regarded as amenable to neural transplantation. However, the ability of NSCs to engraft diffusely and become integral members of structures throughout the host CNS, while also expressing therapeutic molecules, may permit these cells to address that challenge. Intriguingly, while NSCs can be readily engineered to express specified foreign genes, other intrinsic factors appear to emanate spontaneously from NSCs and, in the context of reciprocal donor-host signaling, seem to be capable of neuroprotective and/or neuroregenerative functions. Stem cells additionally have the appealing ability to 'home in' on pathology, even over great distances. Such observations help to advance the idea that NSCs - as a prototype for stem cells from other solid organs - might aid in reconstructing the molecular and cellular milieu of maldeveloped or damaged organs.
proteins
2022-02-25 | Increasing Aspartoacylase in the Central Amygdala: The Common Mechanism of Gastroprotective Effects of Monoamine-Based Antidepressants Against Stress
Monoamine-based antidepressants can prophylactically protect against stress-induced gastric ulcers. Although the central nucleus of amygdala (CeA) has been shown to modulate the severity of stress ulcers, little is known about the molecular mechanisms underlying the gastroprotective effect of this kind of drugs. Here, we first used proton magnetic resonance spectroscopy, a non-invasive tool, to explore the change of neurometabolites of the CeA of rats pretreated with the duloxetine of selective serotonin-norepinephrine reuptake inhibitors during 6 h of water-immersion restraint stress (WIRS). Duloxetine decreased N-acetyl-aspartate/creatine ratio (NAA/creatine) in CeA after WIRS, which was paralleled by the amelioration of gastric lesions. Meanwhile, the gastric ulcer index was negatively correlated with reduced NAA/creatine. Furthermore, the intra-CeA infusion of NAA aggravated WIRS-induced gastric mucosa damage, which suggested the crucial role of reduced NAA. Western blotting was performed to identify the specific enzymes responsible for the change of the contents of NAA at 0.5 h/3 h/6 h after WIRS, considering the preventative gastric protection of duloxetine. The NAA-catabolizing enzyme aspartoacylase (ASPA) was the only enzyme downregulated by 0.5 h WIRS and upregulated by duloxetine. Moreover, overexpressing ASPA in CeA alleviated stress ulcers. Additionally, all of the other three monoamine-based antidepressants, the fluoxetine of selective serotonin reuptake inhibitors, the amitriptyline of tricyclic agents, and the moclobemide of MAOs, increased ASPA expression in CeA. Together, these results indicate that increasing ASPA to hydrolyze NAA in CeA is a common mechanism of gastroprotective effects against stress exerted by monoamine-based antidepressants, and ASPA is a shared target more than monoamine regulation for this kind of drugs.
2021-05-25 | Engineering of a critical membrane-anchored enzyme for high solubility and catalytic activity.
Membrane-associated proteins carry out a wide range of essential cellular functions but the structural characterization needed to understand these functions is dramatically underrepresented in the Protein Data Bank. Producing a soluble, stable and active form of a membrane-associated protein presents formidable challenges, as evidenced by the variety of approaches that have been attempted with a multitude of different membrane proteins to achieve this goal. Aspartate N-acetyltransferase (ANAT) is a membrane-anchored enzyme that performs a critical function, the synthesis of N-acetyl-l-aspartate (NAA), the second most abundant amino acid in the brain. This amino acid is a precursor for a neurotransmitter, and alterations in brain NAA levels have been implicated as a causative effect in Canavan disease and has been suggested to be involved in other neurological disorders. Numerous prior attempts have failed to produce a soluble form of ANAT that is amenable for functional and structural investigations. Through the application of a range of different approaches, including fusion partner constructs, linker modifications, membrane-anchor modifications, and domain truncations, a highly soluble, stable and fully active form of ANAT has now been obtained. Producing this modified enzyme form will accelerate studies aimed at structural characterization and structure-guided inhibitor development.
2014-11-01 | Enhanced brain distribution of modified aspartoacylase
Canavan disease is a fatal neurological disorder caused by defects in the gene that produces the enzyme aspartoacylase.Enzyme replacement therapy can potentially be used to overcome these defects if a stable enzyme form can be produced that can gain access to the appropriate neural cells.Achieving the proper cellular targeting requires a modified form of aspartoacylase that can traverse the blood-brain barrier.A PEGylated form of aspartoacylase has been produced that shows dramatic enhancement in brain tissue access and distribution.While the mechanism of transport has not yet been established, this modified enzyme is significantly less immunogenic than unmodified aspartoacylase.These improved properties set the stage for more extensive enzyme replacement trials as a possible treatment strategy.
2011-02-01 | Modification of aspartoacylase for potential use in enzyme replacement therapy for the treatment of Canavan disease
Canavan disease is a fatal neurological disease without any effective treatments to slow the relentless progress of this disorder. Enzyme replacement therapy has been used effectively to treat a number of metabolic disorders, but the presence of the blood–brain-barrier presents an additional challenge in the treatment of neurological disorders. Studies have begun with the aim of establishing a treatment protocol that can effectively replace the defective enzyme in Canavan disease patients. The human enzyme, aspartoacylase, has been cloned, expressed and purified, and the surface lysyl groups modified through PEGylation. Fully active modified enzymes were administered to mice that are defective in this enzyme and that show many of the symptoms of Canavan disease. Statistically significant increases in brain enzyme activity levels have been achieved in this animal model, as well as decreases in the elevated substrate levels that mimic those found in Canavan disease patients. These results demonstrate that the modified enzyme is gaining access to the brain and functions to correct this metabolic defect. The stage is now set for a long term study to optimize this enzyme replacement approach for the development of a treatment protocol.
2006-04-13 | Characterization of Human Aspartoacylase: The Brain Enzyme Responsible for Canavan Disease
Aspartoacylase catalyzes the deacetylation of N-acetylaspartic acid (NAA) to produce acetate and l-aspartate and is the only brain enzyme that has been shown to effectively metabolize NAA. Although the exact role of this enzymatic reaction has not yet been completely elucidated, the metabolism of NAA appears to be necessary in the formation of myelin lipids, and defects in this enzyme lead to Canavan disease, a fatal neurological disorder. The low catalytic activity and inherent instability observed with the Escherichia coli-expressed form of aspartoacylase suggested the need for a suitable eukaryotic expression system that would be capable of producing a fully functional, mature enzyme. Human aspartoacylase has now been successfully expressed in Pichia pastoris. While the expression yields are lower than in E. coli, the purified enzyme is significantly more stable. This enzyme form has the same substrate specificity but is 150-fold more active than the E. coli-expressed enzyme. The molecular weight of the purified enzyme, measured by mass spectrometry, is higher than predicted, suggesting the presence of some post-translational modifications. Deglycosylation of aspartoacylase or mutation at the glycosylation site causes decreased enzyme stability and diminished catalytic activity. A carbohydrate component has been removed and characterized by mass spectrometry. In addition to this carbohydrate moiety, the enzyme has also been shown to contain one zinc atom per subunit. Chelation studies to remove the zinc result in a reversible loss of catalytic activity, thus establishing aspartoacylase as a zinc metalloenzyme.
gene therapies
2026-07-30 | Infantile-Onset Canavan Disease: A Rare Type of Leukodystrophy
Abstract Canavan disease is a rare form of leucodystrophy due to the deficiency of aspartoacylase enzyme, characertized by global developmental delay, spastic quadriparesis and macrocephaly. The neuroimagimg hallmark is the involvement of periventricular and subcortical white matter and tall N-acetyl aspartic acid peak.
2026-02-23 | The Cell- and Tissue-Type Dependent Activity of the Mouse Aspa Promoter and its Proximal Enhancers
Aspartoacylase (ASPA) is the enzyme responsible for metabolizing n-acetylaspartate, and for which loss-of-function mutations are causal for Canavan disease (CD). Aspa is primarily expressed in oligodendrocytes, but it is also found throughout the body. However, Aspa regulation in most tissues remains poorly explored. We identified putative mouse Aspa regulatory elements in a 3.4 kb sequence upstream of the transcriptional start site using publicly available epigenetic datasets. The sequence includes two putative tissue-specific enhancer sequences, enh1 and enh2, two non-hypersensitive regions, and a proximal promoter. To define the functional architecture of the putative Aspa promoter, we generated a series of promoter mutants with each identified sequence element deleted singly or in series. Plasmid reporter assay experiments in six cell lines derived from distinct tissue types, and in vivo mouse experiments with Aspa promoter AAV9 vectors, allowed the quantification of promoter activity in multiple contexts. We find that in the plasmid and AAV context, enh1 acts as a broadly active silencer, and that enh2 acts as a broadly active enhancer. The Aspa promoter showed minimal activity in low-Aspa expressing cell-types and tissues, including neurons and heart, which has the potential to enhance the safety of future gene therapy treatments which may utilize this promoter. The large enh2 sequence was further found to contain two small and adjacent silencer and enhancer elements. Using dsDNA-affinity-mass spectrometry, we identified CUX1 as a candidate binding protein of the isolated silencer sequence. We tested the effect of CUX1 on Aspa promoter activity using motif deletion, overexpression, and siRNA knockdown strategies. These experiments suggest that CUX1 may act as a repressor of Aspa promoter activity. However, our experiments were unable to confirm that the interaction of CUX1 and the mouse Aspa promoter is direct. In summary, the mouse Aspa promoter recapitulates aspects of endogenous Aspa expression and may be useful in the future as an endogenously derived regulatory sequence for Canavan gene therapy.
2025-10-29 | A Rare Leukodystrophy Revealed in a Moroccan Infant: A Case Report of Canavan Disease
Introduction: Canavan disease is a rare autosomal-recessive leukodystrophy characterized by spongiform degeneration of the cerebral white matter secondary to aspartoacylase (ASPA) deficiency. It presents early with axial hypotonia, global psychomotor delay, and progressive macrocephaly. Objective: This work aims to illustrate the clinical, biological, and radiological particularities of this disorder through a Moroccan case. Case Report: We report the case of a 10-month-old female infant, born to first-degree consanguineous parents, presenting with severe axial hypotonia, absence of head control, generalized seizures, and macrocephaly. Brain MRI showed diffuse, symmetrical T2-weighted hyperintensities of the white matter, typical of spongiform leukodystrophy. Urinary assay revealed marked elevation of N-acetyl-aspartic acid (NAA), and molecular analysis confirmed homozygosity for the c.924del ASPA mutation. Management was symptomatic, including motor rehabilitation and nutritional assistance. Discussion: This observation illustrates the severe infantile form of Canavan disease. The combination of macrocephaly, developmental delay, and diffuse white-matter abnormalities should alert the clinician. Although differential diagnoses include other leukodystrophies, increased NAA and molecular confirmation are specific. Familial consanguinity and the presence of an affected sibling highlight the need for family screening and genetic counseling. Conclusion: Although no curative treatment exists, early identification of this rare disease allows appropriate management and opens perspectives for innovative therapeutic strategies, particularly gene therapy.
2025-09-17 | Oligodendrocyte-targeted adeno-associated virus gene therapy for Canavan disease in children: a phase 1/2 trial.
This open-label phase 1/2 clinical study uses a novel recombinant vector, rAAV-Olig001, with selective tropism for oligodendrocytes, to deliver gene therapy for Canavan disease (CD), a rare leukodystrophy characterized by defective aspartoacylase and elevated N-acetyl-aspartic acid (NAA) concentrations. A total of 8 participants received intracranial doses of 3.7 × 1013 vector genomes (vg) of rAAV-Olig001-ASPA (MYR-101), with an interim analysis at 12 months. The primary objective was to assess the safety of intracranial dosing of MYR-101 in children with typical CD. Efficacy measures included Mullen Scales of Early Learning (MSEL), Gross Motor Function Measure (GMFM) and analysis of NAA, myelination, white matter and extracellular water content in the brain. The participants were White; 5 (62.5%) were male. Of the participants, 7 (87.5%) experienced ≥1 serious adverse event, none of which were considered MYR-101 related. All participants experienced ≥1 adverse event. All adverse events and serious adverse events resolved fully. Treatment reduced NAA concentrations in cerebrospinal fluid (P = 0.0008), increased myelination (P = 0.0137) and improved MSEL developmental outcomes (P = 0.0171). Thus, interim results suggest that gene therapy with MYR-101 is well tolerated and shows early effects in CD. While these findings are preliminary, reductions in NAA concentrations indicate ASPA expression and increases in myelination and imply successful targeting of oligodendrocytes. These results may support the development of similar gene therapy strategies for other demyelinating and metabolic brain disorders. ClinicalTrials.gov registration: NCT04833907 .
2024-08-28 | Neurosurgical gene therapy for central nervous system diseases.
Viral vector mediated gene therapies for neurodegenerative and neurodevelopmental conditions that require neurosurgical administration continue to expand. We systematically reviewed the National Institutes of Health (NIH) ClinicalTrials.gov database to identify all clinical trials studying in-vivo viral vector mediated gene therapies targeted to the CNS for neurodegenerative and neurodevelopmental diseases. We isolated studies which delivered therapies using neurosurgical approaches: intracisternal, intraventricular, and/or intraparenchymal. Clinical trials primarily registered in international countries were included if they were referenced by an NIH registered clinical trial. We performed a scoping review to identify the preclinical studies that supported each human clinical trial. Key preclinical and clinical data were aggregated to characterize vector capsid design, delivery methods, gene expression profile, and clinical benefit. A total of 64 clinical trials were identified in active, completed, terminated, and long-term follow-up stages. A range of CNS conditions across pediatric and adult populations are being studied with CNS targeted viral vector gene therapy, including Alzheimer's disease, Parkinson's disease, AADC deficiency, sphingolipidoses, mucopolysaccharidoses, neuronal ceroid lipofuscinoses, spinal muscular atrophy, adrenoleukodystrophy, Canavan disease, frontotemporal dementia, Huntington's disease, Rett syndrome, Dravet syndrome, mesial temporal lobe epilepsy, and glutaric acidemia. Adeno-associated viral vectors (AAVs) were utilized by the majority of tested therapies, with vector serotypes, regulatory elements, delivery methods, and vector monitoring varying based on the disease being studied. Intraparenchymal delivery has evolved significantly, with MRI-guided convection-enhanced delivery established as a gold standard method for pioneering novel gene targets.
other
2023-10-26 | Pathological Bergmann glia alterations and disrupted calcium dynamics in ataxic Canavan disease mice.
Canavan disease (CD) is a recessively inherited pediatric leukodystrophy resulting from inactivating mutations to the oligodendroglial enzyme aspartoacylase (ASPA). ASPA is responsible for hydrolyzing the amino acid derivative N-acetyl-L-aspartate (NAA), and without it, brain NAA concentrations increase by 50% or more. Infants and children with CD present with progressive cognitive and motor delays, cytotoxic edema, astroglial vacuolation, and prominent spongiform brain degeneration. ASPA-deficient CD mice (Aspanur7/nur7 ) present similarly with elevated NAA, widespread astroglial dysfunction, ataxia, and Purkinje cell (PC) dendritic atrophy. Bergmann glia (BG), radial astrocytes essential for cerebellar development, are intimately intertwined with PCs, where they regulate synapse stability, functionality, and plasticity. BG damage is common to many neurodegenerative conditions and frequently associated with PC dysfunction and ataxia. Here, we report that, in CD mice, BG exhibit significant morphological alterations, decreased structural associations with PCs, loss of synaptic support proteins, and altered calcium dynamics. We also find that BG dysfunction predates cerebellar vacuolation and PC damage in CD mice. Previously, we developed an antisense oligonucleotide (ASO) therapy targeting Nat8l (N-acetyltransferase-8-like, "Nat8l ASO") that inhibits the production of NAA and reverses ataxia and PC atrophy in CD mice. Here, we show that Nat8l ASO administration in adult CD mice also leads to BG repair. Furthermore, blocking astroglial uptake of NAA is neuroprotective in astroglia-neuron cocultures exposed to elevated NAA. Our findings suggest that restoration of BG structural and functional integrity could be a mechanism for PC regeneration and improved motor function.
2020-08-03 | Antisense Oligonucleotide Reverses Leukodystrophy in Canavan Disease Mice.
Marked elevation in the brain concentration of N-acetyl-L-aspartate (NAA) is a characteristic feature of Canavan disease, a vacuolar leukodystrophy resulting from deficiency of the oligodendroglial NAA-cleaving enzyme aspartoacylase. We now demonstrate that inhibiting NAA synthesis by intracisternal administration of a locked nucleic acid antisense oligonucleotide to young-adult aspartoacylase-deficient mice reverses their pre-existing ataxia and diminishes cerebellar and thalamic vacuolation and Purkinje cell dendritic atrophy. Ann Neurol 2020;87:480-485.
small molecules
2026-02-12 | Discovery of N-Acetyltransferase 8-Like (NAT8L) inhibitors based on a N-Acylated (Piperidin-3-ylmethyl)-1,2,4-Oxadiazole Scaffold.
Canavan disease (CD) is an autosomal recessive genetic disorder caused by mutations in the ASPA gene, which encodes the enzyme aspartoacylase. These mutations lead to a deficient enzymatic activity and increased concentrations of its substrate, N-acetylaspartate (NAA), in the brain and other tissues. Aspartate N-acetyltransferase, encoded by the N-acetyltransferase 8-like (NAT8L) gene, catalyzes the biosynthesis of NAA from aspartate and acetyl-CoA. Therefore, inhibition of NAT8L has been implicated as a promising therapeutic strategy for CD by normalizing NAA levels in the brain. Our high throughput screening campaign followed by a rigorous hit validation process identified 2-(2-fluorophenoxy)-1-(3-((3-(thiophen-3-yl)-1,2,4-oxadiazol-5-yl)methyl)piperidin-1-yl)ethan-1-one (4a) as a low micromolar, noncarboxylic acid inhibitor of NAT8L. Subsequent structural optimization led to the discovery of two submicromolar NAT8L inhibitors. Although these inhibitors displayed high clearance in liver microsomes, the new scaffold, devoid of a carboxylic acid moiety, could potentially lead to potent and brain-penetrant NAT8L inhibitors through further molecular refinement.
2024-11-26 | N-acetylaspartate mitigates pro-inflammatory responses in microglial cells by intersecting lipid metabolism and acetylation processes.
Microglia play a crucial role in brain development and repair by facilitating processes such as synaptic pruning and debris clearance. They can be activated in response to various stimuli, leading to either pro-inflammatory or anti-inflammatory responses associated with specific metabolic alterations. The imbalances between microglia activation states contribute to chronic neuroinflammation, a hallmark of neurodegenerative diseases. N-acetylaspartate (NAA) is a brain metabolite predominantly produced by neurons and is crucial for central nervous system health. Alterations in NAA metabolism are observed in disorders such as Multiple Sclerosis and Canavan disease. While NAA's role in oligodendrocytes and astrocytes has been investigated, its impact on microglial function remains less understood. The murine BV2 microglial cell line and primary microglia were used as experimental models. Cells were treated with exogenous NAA and stimulated with LPS/IFN-γ to reproduce the pro-inflammatory phenomenon. HPLC and immunofluorescence analysis were used to study lipid metabolism following NAA treatment. Automated fluorescence microscopy was used to analyze phagocytic activity. The effects on the pro-inflammatory response were evaluated by analysis of protein/mRNA expression and ChIP assay of typical inflammatory markers. NAA treatment promotes an increase in both lipid synthesis and degradation, and enhances the phagocytic activity of BV2 cells, thus fostering surveillant microglia characteristics. Importantly, NAA decreases the pro-inflammatory state induced by LPS/IFN-γ via the activation of histone deacetylases (HDACs). These findings were validated in primary microglial cells, highlighting the impact on cellular metabolism and inflammatory responses. The study highlighted the role of NAA in reinforcing the oxidative metabolism of surveillant microglial cells and, most importantly, in buffering the inflammatory processes characterizing reactive microglia. These results suggest that the decreased levels of NAA observed in neurodegenerative disorders can contribute to chronic neuroinflammation.
2024-11-14 | Restoring Function in Pediatric Neurodegeneration: The Impact of Radio Electric Asymmetric Conveyor Neuroregenerative Treatment in a Child With Canavan Syndrome.
This case report presents the therapeutic impact of radio electric asymmetric conveyor (REAC) neuroregenerative medicine treatment (REAC RGN-N) in a 10-year-old girl diagnosed with Canavan syndrome, a rare genetic disorder marked by progressive white matter degeneration in the brain. This condition, caused by aspartoacylase deficiency, leads to an accumulation of N-acetyl-L-aspartic acid (NAA), resulting in severe motor impairment, muscle tone abnormalities, and developmental delays. The patient received REAC RGN-N treatment, administered via the REAC - BENE mod 110 (ASMED®, Scandicci, Italy), specifically configured for RGN-N therapy. The protocol consisted of four 6-hour sessions completed over a 24-hour cycle. Following this initial treatment, the patient showed notable improvements in head and trunk control, muscle tone, and swallowing function. Head and trunk control increased from 20 to 30 on the trunk control measurement scale (TCMS), spasticity decreased from a score of 3 to 1 on the Ashworth scale, and her swallowing ability progressed from level 5 to level 7 on the functional oral intake scale (FOIS), allowing her to handle a wider range of food textures independently. These improvements were assessed six months after the end of the first treatment cycle, suggesting a degree of stability in the therapeutic effects observed. Although further studies could support these findings, the rarity of Canavan syndrome presents challenges for conducting additional targeted research. REAC RGN-N treatment is typically repeated in cycles every 4-6 months, depending on clinical severity, to sustain therapeutic benefits. This case report offers initial evidence that REAC RGN-N may provide meaningful, stable improvements for pediatric patients with neurodegenerative conditions, addressing motor and sensory impairments with potential long-term benefits.
2024-08-12 | Identification of potential pharmacological chaperones that selectively stabilize mutated Aspartoacylases in Canavan disease.
Canavan disease is caused by mutations in the ASPA gene, leading to diminished catalytic activity of aspartoacylase in the brain. Clinical missense mutations are found throughout the enzyme structure, with many of these mutated enzymes having not only decreased activity but also compromised stability. High-throughput screening of a small molecule library has identified several compounds that significantly increase the thermal stability of the E285A mutant enzyme, the most predominant clinical mutation in Canavan disease, while having a negligible effect on the native enzyme. Based on the initial successes, some structural analogs of these initial hits were selected for further examination. Glutathione, NAAG and patulin were each confirmed to be competitive inhibitors, indicating the binding of these compounds at the dimer interface or near the active site of the E285A enzyme. The experimental results were theoretically examined with the help of the docking analysis method. The structure activity-guided optimization of these compounds can potentially lead to potential pharmacological chaperones that could alleviate the detrimental effect of ASPA mutations in Canavan patients.
2024-01-28 | Astroglial conditional Slc13a3 knockout is therapeutic in murine Canavan leukodystrophy.
Abstract Canavan disease is a leukodystrophy caused by ASPA mutations that diminish oligodendroglial aspartoacylase activity, and is characterized by markedly elevated brain concentrations of the aspartoacylase substrate N‐acetyl‐ l ‐aspartate (NAA) and by astroglial and intramyelinic vacuolation. Astroglia express NaDC3 (encoded by SLC13A3 ), a sodium‐coupled transporter for NAA and other dicarboxylates. Astroglial conditional Slc13a3 deletion in aspartoacylase‐deficient Canavan disease model mice (“CD mice”) reversed brain NAA elevation and improved motor function. These results demonstrate that astroglial NaDC3 contributes to brain NAA elevation in CD mice, and suggest that suppressing astroglial NaDC3 activity would ameliorate human Canavan disease.
cell therapies
2026-01-24 | Human iPSC-derived neural progenitor cells rescue motor function and brain pathology in symptomatic Canavan disease mice.
Canavan disease (CD) is a severe neurodegenerative disorder caused by aspartoacylase (ASPA) deficiency, leading to N-acetyl-L-aspartic acid (NAA) accumulation and spongy degeneration. While several therapeutic candidates improve outcomes in CD mouse models when delivered before symptom onset, there remains a need for treatments targeting established disease pathology. Here, we demonstrate that transplantation with human induced pluripotent stem cell (iPSC)-derived neural progenitor cells (NPCs) expressing a functional ASPA gene (ASPA iNPCs) can rescue disease manifestations in symptomatic CD (Nur7) mice. When administered at postnatal day 21 (P21), ASPA iNPCs successfully engrafted, differentiated into neural lineage cells, and restored ASPA activity as revealed by reduced NAA level. Transplanted mice showed a significant reduction in brain and cerebrospinal fluid (CSF) NAA levels, decreased vacuolation across multiple brain regions, improved myelination, and enhanced motor function 6-month post-transplantation. Our findings demonstrate that ASPA iNPC transplantation can effectively reverse established CD pathology, suggesting therapeutic potential for treating symptomatic patients.
2023-06-04 | Developing Hypoimmunogenic Human iPSC‐Derived Oligodendrocyte Progenitor Cells as an Off‐The‐Shelf Cell Therapy for Myelin Disorders
Demyelinating disorders are among the most common and debilitating diseases in neurology. Canavan disease (CD) is a lethal demyelinating disease caused by mutation of the aspartoacylase (ASPA) gene, which leads to the accumulation of its substrate N-acetyl-l-aspartate (NAA), and consequently demyelination and vacuolation in the brain. In this study, hypoimmunogenic human induced pluripotent stem cell (iPSC)-derived oligodendrocyte progenitor cells (OPC) are developed from a healthy donor as an "off-the-shelf" cell therapy. Hypoimmunogenic iPSCs are generated through CRISPR/Cas9 editing of the human leukocyte antigen (HLA) molecules in healthy donor-derived iPSCs and differentiated into OPCs. The OPCs are engrafted into the brains of CD (nur7) mice and exhibit widespread distribution in the brain. The engrafted OPCs mature into oligodendrocytes that express the endogenous wildtype ASPA gene. Consequently, the transplanted mice exhibit elevated human ASPA expression and enzymatic activity and reduced NAA level in the brain. The transplanted OPCs are able to rescue major pathological features of CD, including defective myelination, extensive vacuolation, and motor function deficits. Moreover, the hypoimmunogenic OPCs exhibit low immunogenicity both in vitro and in vivo. The hypoimmunogenic OPCs can be used as "off-the-shelf" universal donor cells to treat various CD patients and many other demyelinating disorders, especially autoimmune demyelinating diseases, such as multiple sclerosis.
2014-01-06 | Advances and pitfalls of cell therapy in metabolic leukodystrophies.
Leukodystrophies are a group of disorders characterized by myelin dysfunction, either at the level of myelin formation or maintenance, that affect the central nervous system (CNS) and also in some cases, to a lesser extent, the peripheral nervous system (PNS). Although these genetic-based disorders are generally rare, all together they have a significant impact in the society, with an estimated overall incidence of 1 in 7,663 live births. Currently, there is no cure for leukodystrophies, and the development of effective treatments remains challenging. Not only leukodystrophies generally progress very fast, but also most are multifocal needing the simultaneous targeting at multiple sites. Moreover, as the CNS is affected, the blood-brain barrier (BBB) limits the efficacy of treatment. Recently, interest on cell therapy has increased, and the leukodystrophies for which metabolic correction is needed have become first-choice candidates for cell-based clinical trials. In this review, we present and discuss the available cell transplantation therapies in metabolic leukodystrophies including fucosidosis, X-linked adrenoleukodystrophy, metachromatic leukodystrophy, Canavan disease, and Krabbe's disease. We will discuss the latest advances of cell therapy and its pitfalls in this group of disorders, taking into account, among others, the limitations imposed by reduced cell migration in multifocal conditions, the need to achieve corrective enzyme threshold levels, and the growing awareness that not only myelin but also the associated axonopathy needs to be targeted in some leukodystrophies.
2004-10-01 | Mouse neural progenitor cells differentiate into oligodendrocytes in the brain of a knockout mouse model of Canavan disease
Canavan disease (CD) is an autosomal recessive disorder that leads to spongy degeneration in the white matter of the brain. Aspartoacylase (ASPA) synthesizing cells, oligodendrocytes, are lost in CD. Transplantation of neural progenitor cells (NPCs) offers an interesting therapeutic approach for treating neurodegenerative diseases by replacing the lost cells. Therefore, the NPCs transplantation to the brain of the CD mouse was studied. Injection of mouse NPCs to the striatum and cerebellum of juvenile CD mouse showed numerous BrdU positive cells at 1 month after injection. The same result was also observed in the adult CD mouse brain after 5 weeks of post-transplantation period. The implanted cells differentiated into oligodendrocytes and fibrous astrocytes, as observed using glial cell marker. This is the first report to describe the survival, distribution and differentiation of NPCs within the brain of CD mouse and a first step toward the potential clinical use of cell therapy to treat CD.
2002-05-01 | Global gene and cell replacement strategies via stem cells
The inherent biology of neural stem cells (NSCs) endows them with capabilities that not only circumvent many of the limitations of other gene transfer vehicles, but that enable a variety of novel therapeutic strategies heretofore regarded as beyond the purview of neural transplantation. Most neurodegenerative diseases are characterized not by discrete, focal abnormalities but rather by extensive, multifocal, or even global neuropathology. Such widely disseminated lesions have not conventionally been regarded as amenable to neural transplantation. However, the ability of NSCs to engraft diffusely and become integral members of structures throughout the host CNS, while also expressing therapeutic molecules, may permit these cells to address that challenge. Intriguingly, while NSCs can be readily engineered to express specified foreign genes, other intrinsic factors appear to emanate spontaneously from NSCs and, in the context of reciprocal donor-host signaling, seem to be capable of neuroprotective and/or neuroregenerative functions. Stem cells additionally have the appealing ability to 'home in' on pathology, even over great distances. Such observations help to advance the idea that NSCs - as a prototype for stem cells from other solid organs - might aid in reconstructing the molecular and cellular milieu of maldeveloped or damaged organs.
proteins
2022-02-25 | Increasing Aspartoacylase in the Central Amygdala: The Common Mechanism of Gastroprotective Effects of Monoamine-Based Antidepressants Against Stress
Monoamine-based antidepressants can prophylactically protect against stress-induced gastric ulcers. Although the central nucleus of amygdala (CeA) has been shown to modulate the severity of stress ulcers, little is known about the molecular mechanisms underlying the gastroprotective effect of this kind of drugs. Here, we first used proton magnetic resonance spectroscopy, a non-invasive tool, to explore the change of neurometabolites of the CeA of rats pretreated with the duloxetine of selective serotonin-norepinephrine reuptake inhibitors during 6 h of water-immersion restraint stress (WIRS). Duloxetine decreased N-acetyl-aspartate/creatine ratio (NAA/creatine) in CeA after WIRS, which was paralleled by the amelioration of gastric lesions. Meanwhile, the gastric ulcer index was negatively correlated with reduced NAA/creatine. Furthermore, the intra-CeA infusion of NAA aggravated WIRS-induced gastric mucosa damage, which suggested the crucial role of reduced NAA. Western blotting was performed to identify the specific enzymes responsible for the change of the contents of NAA at 0.5 h/3 h/6 h after WIRS, considering the preventative gastric protection of duloxetine. The NAA-catabolizing enzyme aspartoacylase (ASPA) was the only enzyme downregulated by 0.5 h WIRS and upregulated by duloxetine. Moreover, overexpressing ASPA in CeA alleviated stress ulcers. Additionally, all of the other three monoamine-based antidepressants, the fluoxetine of selective serotonin reuptake inhibitors, the amitriptyline of tricyclic agents, and the moclobemide of MAOs, increased ASPA expression in CeA. Together, these results indicate that increasing ASPA to hydrolyze NAA in CeA is a common mechanism of gastroprotective effects against stress exerted by monoamine-based antidepressants, and ASPA is a shared target more than monoamine regulation for this kind of drugs.
2021-05-25 | Engineering of a critical membrane-anchored enzyme for high solubility and catalytic activity.
Membrane-associated proteins carry out a wide range of essential cellular functions but the structural characterization needed to understand these functions is dramatically underrepresented in the Protein Data Bank. Producing a soluble, stable and active form of a membrane-associated protein presents formidable challenges, as evidenced by the variety of approaches that have been attempted with a multitude of different membrane proteins to achieve this goal. Aspartate N-acetyltransferase (ANAT) is a membrane-anchored enzyme that performs a critical function, the synthesis of N-acetyl-l-aspartate (NAA), the second most abundant amino acid in the brain. This amino acid is a precursor for a neurotransmitter, and alterations in brain NAA levels have been implicated as a causative effect in Canavan disease and has been suggested to be involved in other neurological disorders. Numerous prior attempts have failed to produce a soluble form of ANAT that is amenable for functional and structural investigations. Through the application of a range of different approaches, including fusion partner constructs, linker modifications, membrane-anchor modifications, and domain truncations, a highly soluble, stable and fully active form of ANAT has now been obtained. Producing this modified enzyme form will accelerate studies aimed at structural characterization and structure-guided inhibitor development.
2014-11-01 | Enhanced brain distribution of modified aspartoacylase
Canavan disease is a fatal neurological disorder caused by defects in the gene that produces the enzyme aspartoacylase.Enzyme replacement therapy can potentially be used to overcome these defects if a stable enzyme form can be produced that can gain access to the appropriate neural cells.Achieving the proper cellular targeting requires a modified form of aspartoacylase that can traverse the blood-brain barrier.A PEGylated form of aspartoacylase has been produced that shows dramatic enhancement in brain tissue access and distribution.While the mechanism of transport has not yet been established, this modified enzyme is significantly less immunogenic than unmodified aspartoacylase.These improved properties set the stage for more extensive enzyme replacement trials as a possible treatment strategy.
2011-02-01 | Modification of aspartoacylase for potential use in enzyme replacement therapy for the treatment of Canavan disease
Canavan disease is a fatal neurological disease without any effective treatments to slow the relentless progress of this disorder. Enzyme replacement therapy has been used effectively to treat a number of metabolic disorders, but the presence of the blood–brain-barrier presents an additional challenge in the treatment of neurological disorders. Studies have begun with the aim of establishing a treatment protocol that can effectively replace the defective enzyme in Canavan disease patients. The human enzyme, aspartoacylase, has been cloned, expressed and purified, and the surface lysyl groups modified through PEGylation. Fully active modified enzymes were administered to mice that are defective in this enzyme and that show many of the symptoms of Canavan disease. Statistically significant increases in brain enzyme activity levels have been achieved in this animal model, as well as decreases in the elevated substrate levels that mimic those found in Canavan disease patients. These results demonstrate that the modified enzyme is gaining access to the brain and functions to correct this metabolic defect. The stage is now set for a long term study to optimize this enzyme replacement approach for the development of a treatment protocol.
2006-04-13 | Characterization of Human Aspartoacylase: The Brain Enzyme Responsible for Canavan Disease
Aspartoacylase catalyzes the deacetylation of N-acetylaspartic acid (NAA) to produce acetate and l-aspartate and is the only brain enzyme that has been shown to effectively metabolize NAA. Although the exact role of this enzymatic reaction has not yet been completely elucidated, the metabolism of NAA appears to be necessary in the formation of myelin lipids, and defects in this enzyme lead to Canavan disease, a fatal neurological disorder. The low catalytic activity and inherent instability observed with the Escherichia coli-expressed form of aspartoacylase suggested the need for a suitable eukaryotic expression system that would be capable of producing a fully functional, mature enzyme. Human aspartoacylase has now been successfully expressed in Pichia pastoris. While the expression yields are lower than in E. coli, the purified enzyme is significantly more stable. This enzyme form has the same substrate specificity but is 150-fold more active than the E. coli-expressed enzyme. The molecular weight of the purified enzyme, measured by mass spectrometry, is higher than predicted, suggesting the presence of some post-translational modifications. Deglycosylation of aspartoacylase or mutation at the glycosylation site causes decreased enzyme stability and diminished catalytic activity. A carbohydrate component has been removed and characterized by mass spectrometry. In addition to this carbohydrate moiety, the enzyme has also been shown to contain one zinc atom per subunit. Chelation studies to remove the zinc result in a reversible loss of catalytic activity, thus establishing aspartoacylase as a zinc metalloenzyme.
gene therapies
2026-07-30 | Infantile-Onset Canavan Disease: A Rare Type of Leukodystrophy
Abstract Canavan disease is a rare form of leucodystrophy due to the deficiency of aspartoacylase enzyme, characertized by global developmental delay, spastic quadriparesis and macrocephaly. The neuroimagimg hallmark is the involvement of periventricular and subcortical white matter and tall N-acetyl aspartic acid peak.
2026-02-23 | The Cell- and Tissue-Type Dependent Activity of the Mouse Aspa Promoter and its Proximal Enhancers
Aspartoacylase (ASPA) is the enzyme responsible for metabolizing n-acetylaspartate, and for which loss-of-function mutations are causal for Canavan disease (CD). Aspa is primarily expressed in oligodendrocytes, but it is also found throughout the body. However, Aspa regulation in most tissues remains poorly explored. We identified putative mouse Aspa regulatory elements in a 3.4 kb sequence upstream of the transcriptional start site using publicly available epigenetic datasets. The sequence includes two putative tissue-specific enhancer sequences, enh1 and enh2, two non-hypersensitive regions, and a proximal promoter. To define the functional architecture of the putative Aspa promoter, we generated a series of promoter mutants with each identified sequence element deleted singly or in series. Plasmid reporter assay experiments in six cell lines derived from distinct tissue types, and in vivo mouse experiments with Aspa promoter AAV9 vectors, allowed the quantification of promoter activity in multiple contexts. We find that in the plasmid and AAV context, enh1 acts as a broadly active silencer, and that enh2 acts as a broadly active enhancer. The Aspa promoter showed minimal activity in low-Aspa expressing cell-types and tissues, including neurons and heart, which has the potential to enhance the safety of future gene therapy treatments which may utilize this promoter. The large enh2 sequence was further found to contain two small and adjacent silencer and enhancer elements. Using dsDNA-affinity-mass spectrometry, we identified CUX1 as a candidate binding protein of the isolated silencer sequence. We tested the effect of CUX1 on Aspa promoter activity using motif deletion, overexpression, and siRNA knockdown strategies. These experiments suggest that CUX1 may act as a repressor of Aspa promoter activity. However, our experiments were unable to confirm that the interaction of CUX1 and the mouse Aspa promoter is direct. In summary, the mouse Aspa promoter recapitulates aspects of endogenous Aspa expression and may be useful in the future as an endogenously derived regulatory sequence for Canavan gene therapy.
2025-10-29 | A Rare Leukodystrophy Revealed in a Moroccan Infant: A Case Report of Canavan Disease
Introduction: Canavan disease is a rare autosomal-recessive leukodystrophy characterized by spongiform degeneration of the cerebral white matter secondary to aspartoacylase (ASPA) deficiency. It presents early with axial hypotonia, global psychomotor delay, and progressive macrocephaly. Objective: This work aims to illustrate the clinical, biological, and radiological particularities of this disorder through a Moroccan case. Case Report: We report the case of a 10-month-old female infant, born to first-degree consanguineous parents, presenting with severe axial hypotonia, absence of head control, generalized seizures, and macrocephaly. Brain MRI showed diffuse, symmetrical T2-weighted hyperintensities of the white matter, typical of spongiform leukodystrophy. Urinary assay revealed marked elevation of N-acetyl-aspartic acid (NAA), and molecular analysis confirmed homozygosity for the c.924del ASPA mutation. Management was symptomatic, including motor rehabilitation and nutritional assistance. Discussion: This observation illustrates the severe infantile form of Canavan disease. The combination of macrocephaly, developmental delay, and diffuse white-matter abnormalities should alert the clinician. Although differential diagnoses include other leukodystrophies, increased NAA and molecular confirmation are specific. Familial consanguinity and the presence of an affected sibling highlight the need for family screening and genetic counseling. Conclusion: Although no curative treatment exists, early identification of this rare disease allows appropriate management and opens perspectives for innovative therapeutic strategies, particularly gene therapy.
2025-09-17 | Oligodendrocyte-targeted adeno-associated virus gene therapy for Canavan disease in children: a phase 1/2 trial.
This open-label phase 1/2 clinical study uses a novel recombinant vector, rAAV-Olig001, with selective tropism for oligodendrocytes, to deliver gene therapy for Canavan disease (CD), a rare leukodystrophy characterized by defective aspartoacylase and elevated N-acetyl-aspartic acid (NAA) concentrations. A total of 8 participants received intracranial doses of 3.7 × 1013 vector genomes (vg) of rAAV-Olig001-ASPA (MYR-101), with an interim analysis at 12 months. The primary objective was to assess the safety of intracranial dosing of MYR-101 in children with typical CD. Efficacy measures included Mullen Scales of Early Learning (MSEL), Gross Motor Function Measure (GMFM) and analysis of NAA, myelination, white matter and extracellular water content in the brain. The participants were White; 5 (62.5%) were male. Of the participants, 7 (87.5%) experienced ≥1 serious adverse event, none of which were considered MYR-101 related. All participants experienced ≥1 adverse event. All adverse events and serious adverse events resolved fully. Treatment reduced NAA concentrations in cerebrospinal fluid (P = 0.0008), increased myelination (P = 0.0137) and improved MSEL developmental outcomes (P = 0.0171). Thus, interim results suggest that gene therapy with MYR-101 is well tolerated and shows early effects in CD. While these findings are preliminary, reductions in NAA concentrations indicate ASPA expression and increases in myelination and imply successful targeting of oligodendrocytes. These results may support the development of similar gene therapy strategies for other demyelinating and metabolic brain disorders. ClinicalTrials.gov registration: NCT04833907 .
2024-08-28 | Neurosurgical gene therapy for central nervous system diseases.
Viral vector mediated gene therapies for neurodegenerative and neurodevelopmental conditions that require neurosurgical administration continue to expand. We systematically reviewed the National Institutes of Health (NIH) ClinicalTrials.gov database to identify all clinical trials studying in-vivo viral vector mediated gene therapies targeted to the CNS for neurodegenerative and neurodevelopmental diseases. We isolated studies which delivered therapies using neurosurgical approaches: intracisternal, intraventricular, and/or intraparenchymal. Clinical trials primarily registered in international countries were included if they were referenced by an NIH registered clinical trial. We performed a scoping review to identify the preclinical studies that supported each human clinical trial. Key preclinical and clinical data were aggregated to characterize vector capsid design, delivery methods, gene expression profile, and clinical benefit. A total of 64 clinical trials were identified in active, completed, terminated, and long-term follow-up stages. A range of CNS conditions across pediatric and adult populations are being studied with CNS targeted viral vector gene therapy, including Alzheimer's disease, Parkinson's disease, AADC deficiency, sphingolipidoses, mucopolysaccharidoses, neuronal ceroid lipofuscinoses, spinal muscular atrophy, adrenoleukodystrophy, Canavan disease, frontotemporal dementia, Huntington's disease, Rett syndrome, Dravet syndrome, mesial temporal lobe epilepsy, and glutaric acidemia. Adeno-associated viral vectors (AAVs) were utilized by the majority of tested therapies, with vector serotypes, regulatory elements, delivery methods, and vector monitoring varying based on the disease being studied. Intraparenchymal delivery has evolved significantly, with MRI-guided convection-enhanced delivery established as a gold standard method for pioneering novel gene targets.
other
2023-10-26 | Pathological Bergmann glia alterations and disrupted calcium dynamics in ataxic Canavan disease mice.
Canavan disease (CD) is a recessively inherited pediatric leukodystrophy resulting from inactivating mutations to the oligodendroglial enzyme aspartoacylase (ASPA). ASPA is responsible for hydrolyzing the amino acid derivative N-acetyl-L-aspartate (NAA), and without it, brain NAA concentrations increase by 50% or more. Infants and children with CD present with progressive cognitive and motor delays, cytotoxic edema, astroglial vacuolation, and prominent spongiform brain degeneration. ASPA-deficient CD mice (Aspanur7/nur7 ) present similarly with elevated NAA, widespread astroglial dysfunction, ataxia, and Purkinje cell (PC) dendritic atrophy. Bergmann glia (BG), radial astrocytes essential for cerebellar development, are intimately intertwined with PCs, where they regulate synapse stability, functionality, and plasticity. BG damage is common to many neurodegenerative conditions and frequently associated with PC dysfunction and ataxia. Here, we report that, in CD mice, BG exhibit significant morphological alterations, decreased structural associations with PCs, loss of synaptic support proteins, and altered calcium dynamics. We also find that BG dysfunction predates cerebellar vacuolation and PC damage in CD mice. Previously, we developed an antisense oligonucleotide (ASO) therapy targeting Nat8l (N-acetyltransferase-8-like, "Nat8l ASO") that inhibits the production of NAA and reverses ataxia and PC atrophy in CD mice. Here, we show that Nat8l ASO administration in adult CD mice also leads to BG repair. Furthermore, blocking astroglial uptake of NAA is neuroprotective in astroglia-neuron cocultures exposed to elevated NAA. Our findings suggest that restoration of BG structural and functional integrity could be a mechanism for PC regeneration and improved motor function.
2020-08-03 | Antisense Oligonucleotide Reverses Leukodystrophy in Canavan Disease Mice.
Marked elevation in the brain concentration of N-acetyl-L-aspartate (NAA) is a characteristic feature of Canavan disease, a vacuolar leukodystrophy resulting from deficiency of the oligodendroglial NAA-cleaving enzyme aspartoacylase. We now demonstrate that inhibiting NAA synthesis by intracisternal administration of a locked nucleic acid antisense oligonucleotide to young-adult aspartoacylase-deficient mice reverses their pre-existing ataxia and diminishes cerebellar and thalamic vacuolation and Purkinje cell dendritic atrophy. Ann Neurol 2020;87:480-485.
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Drug Discovery Landscape
4 orphan drug designations for Canavan disease.
4 orphan drug designations for Canavan disease.
Drug | Therapy type | Orphan designation | Clinical trial status | Sponsor |
|---|---|---|---|---|
Recombinant adeno-associated virus Olig001 containing human aspartoacylase cDNA | gene therapies | EMA | Voisin Consulting Life Sciences | |
recombinant adeno-associated virus containing human ASPA cDNA (rAAV-Olig001-ASPA) | gene therapies | FDA | Myrtelle, Inc. | |
Adeno-associated virus serotype 9 containing the human ASPA gene | gene therapies | EMA | Raremoon Consulting Esp S.L. | |
Self complimentary adeno-associated virus serotype 9 expressing human aspartoacylase transgene | gene therapies | FDA | Aspa Therapeutics, Inc. |
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