Mutations In Transmembrane Protein Research Articles

Abstract 12450: Identifying Differential Proteins of the Arrhythmogenic Right Ventricular Cardiomyopathy Type 5 Associated Mutation in Transmembrane Protein 43 Ser358Leu by Proteomics and Interaction Studies Using TurboID

Introduction: Transmembrane protein 43 (TMEM43) is localized in the nuclear envelope and endoplasmic reticulum (ER). TMEM43- c.1073C>T, p.S358L leads to a rare fully penetrant type 5 arrhythmogenic right ventricular cardiomyopathy (ARVC5). The molecular pathomechanism of ARVC5 is not well understood. Methods: TurboID cloned to N- and C-terminal of TMEM43 WT and p.S358L were transfected in HEK293 cells. Biotinylation assay was performed and biotinylated proteins were immunoprecipitated using streptavidin-beads. After on beads-digestion, peptide fragments were analyzed by Orbitrap mass-spectrometry (MS). Furthermore, we cultivated primary dermal fibroblasts of three TMEM43 p.S358L carriers. Proteins were isolated from cell cultures and analyzed by MS. Additionally, TMEM43-p.S358L and control fibroblasts were investigated by transmission electron microscopy (TEM). Results: TurboID proteome data shows differences in the interacting proteome of TMEM43 WT and p.S358L. VDAC2, which is involved in metabolic regulation of lipids and part of the ER-mitochondrion contact site, was one of the prominent interacting proteins less represented in TMEM43 p.S358L. The proteome of fibroblasts show 11 proteins upregulated in TMEM43-p.S358L (p<0.003). Of note, proteins involved in fatty acid metabolism, such as acyl-coenzym A thioesterase 1 encoded by the gene ACOT1, and Malonate-CoA ligase encoded by ACSF3 were strongly upregulated. ACOT1 and ACSF3 regulate intracellular levels of free fatty acids and are expressed in human cardiomyocytes. TEM images of TMEM43 p.S358L carriers show accumulation of multilamellar bodies, which may support the dysregulation of the lipid metabolism as a prominent pathomechanism. Conclusions: The altered interaction of TMEM43 p.S358L with VDAC2 is a plausible mechanism for miscommunication of ER and mitochondria perturbing the lipid homeostasis. Additionally, the differentially regulated proteins in dermal fibroblasts and the accumulation of multilamellar bodies also suggest a dysregulation of the lipid metabolism by TMEM43 p.S358L, which may explain the development of myocardial fibrofatty replacement contributing to ARVC5.

A mutation in transmembrane protein 135 impairs lipid metabolism in mouse eyecups

Aging is a significant factor in the development of age-related diseases but how aging disrupts cellular homeostasis to cause age-related retinal disease is unknown. Here, we further our studies on transmembrane protein 135 (Tmem135), a gene involved in retinal aging, by examining the transcriptomic profiles of wild-type, heterozygous and homozygous Tmem135 mutant posterior eyecup samples through RNA sequencing (RNA-Seq). We found significant gene expression changes in both heterozygous and homozygous Tmem135 mutant mouse eyecups that correlate with visual function deficits. Further analysis revealed that expression of many genes involved in lipid metabolism are changed due to the Tmem135 mutation. Consistent with these changes, we found increased lipid accumulation in mutant Tmem135 eyecup samples. Since mutant Tmem135 mice have similar ocular pathologies as human age-related macular degeneration (AMD) eyes, we compared our homozygous Tmem135 mutant eyecup RNA-Seq dataset with transcriptomic datasets of human AMD donor eyes. We found similar changes in genes involved in lipid metabolism between the homozygous Tmem135 mutant eyecups and AMD donor eyes. Our study suggests that the Tmem135 mutation affects lipid metabolism as similarly observed in human AMD eyes, thus Tmem135 mutant mice can serve as a good model for the role of dysregulated lipid metabolism in AMD.

MutTMPredictor: Robust and accurate cascade XGBoost classifier for prediction of mutations in transmembrane proteins

Transmembrane proteins have critical biological functions and play a role in a multitude of cellular processes including cell signaling, transport of molecules and ions across membranes. Approximately 60% of transmembrane proteins are considered as drug targets. Missense mutations in such proteins can lead to many diverse diseases and disorders, such as neurodegenerative diseases and cystic fibrosis. However, there are limited studies on mutations in transmembrane proteins. In this work, we first design a new feature encoding method, termed weight attenuation position-specific scoring matrix (WAPSSM), which builds upon the protein evolutionary information. Then, we propose a new mutation prediction algorithm (cascade XGBoost) by leveraging the idea learned from consensus predictors and gcForest. Multi-level experiments illustrate the effectiveness of WAPSSM and cascade XGBoost algorithms. Finally, based on WAPSSM and other three types of features, in combination with the cascade XGBoost algorithm, we develop a new transmembrane protein mutation predictor, named MutTMPredictor. We benchmark the performance of MutTMPredictor against several existing predictors on seven datasets. On the 546 mutations dataset, MutTMPredictor achieves the accuracy (ACC) of 0.9661 and the Matthew’s Correlation Coefficient (MCC) of 0.8950. While on the 67,584 dataset, MutTMPredictor achieves an MCC of 0.7523 and area under curve (AUC) of 0.8746, which are 0.1625 and 0.0801 respectively higher than those of the existing best predictor (fathmm). Besides, MutTMPredictor also outperforms two specific predictors on the Pred-MutHTP datasets. The results suggest that MutTMPredictor can be used as an effective method for predicting and prioritizing missense mutations in transmembrane proteins. The MutTMPredictor webserver and datasets are freely accessible at http://csbio.njust.edu.cn/bioinf/muttmpredictor/ for academic use.

A novel mutation in transmembrane protein 168 causes fatal ventricular arrhythmogenesis in Brugada syndrome

Abstract Brugada syndrome (BrS) is diagnosed by a typical electrocardiography (ECG) with ST-segment elevation in precordial leads and tends to induce sudden cardiac death (SCD) due to ventricular tachycardia/fibrillation. About 20% of SCDs in non-structural cardiac diseases are considered to be caused by BrS. In patients with BrS, loss of function mutations in the Na+ channel is often observed, but the causative gene mutation is not detected for about 70% of BrS patients. Here, we investigated a family with clinically diagnosed BrS, in which no known gene mutations related to BrS had not been found, by whole exome sequencing. Novel heterozygous variant (c. 1616G>A, p. R539Q) in transmembrane protein 168 (TMEM168) was identified only in symptomatic family members. Similar to endogenous TMEM168, both wild-type and mutant TMEM168 localized at the nuclear membrane. Na+ current density in whole-cell patch-clamp recordings was significantly reduced in HL-1 cardiomyocytes transfected with TMEM168 R539Q mutant, compared with those with wild-type TMEM168. Next, heterozygous Tmem168 1616G>A knock-in mice were generated by the CRISPR/Cas9 genome editing technology. Although the knock-in mice had no abnormalities in ECG at the physiological state, the treatment with ajmaline caused various arrhythmias including ventricular tachycardia/fibrillation in the knock-in mice, but not in wild-type mice. Na+ current density and the parameters of action potentials were remarkably impaired in the cardiomyocytes of the knock-in mice. Optical mapping analysis in the whole heart showed the reduced left ventricular conduction velocity in the knock-in mice. The expression of Nav1.5, an α-subunit of the cardiac Na+ channel, was significantly decreased in the mutant TMEM168-transfected HL-1 cells and the knock-in hearts. We found that the decrease was caused by the enhanced ubiquitination of Nav1.5, which was mediated by increased binding of Nedd4–2 E3 ubiquitin ligase to Nav1.5 in the knock-in hearts. Co-immunoprecipitation experiments demonstrated that overactivity of Nedd4–2 is a result of Tmem168 mutant-mediated sequestration of a chaperon protein αB-crystallin, a Nav1.5-binding molecule that interferes with the interaction of Nedd4–2 with Nav1.5. These findings reveal the molecular mechanism of TMEM168 R539Q mutation-induced fatal ventricular arrhythmias in BrS. Funding Acknowledgement Type of funding source: Public Institution(s). Main funding source(s): JSPS Grants-in-aid for Scientific Research

Transmembrane Protein 230 Mediates a Poly(ADP-ribose) Polymerase-1-Linked Apoptosis.

Mutations in transmembrane protein 230 (TMEM230) gene are suggested to be associated with the autosomal dominant Parkinson’s disease (PD) with typical movement disorders and Lewy body pathology. However, the normal functions and the pathological roles of TMEM230 are not clear. In this study, we used TMEM230 isoform II constructs including wild-type (WT) and four reported PD-linked mutation constructs (Y92C, R141L, 184Wext*5, and 184PGext*5). Ectopic expression of WT and PD-linked mutant TMEM230 variants in cultured cells dramatically induced apoptotic cell death compared with that of vector control cells. Mutant TMEM230 caused cell toxicity at an increased severity than WT TMEM230. Moreover, expression of TMEM230 increased mitochondrial reactive oxygen species (ROS) levels, decreased cellular ATP, activated caspase 3/7, and increased poly(ADP-ribose) polymerase-1 (PARP1) cleavage. Treatment with N-acetylcysteine (NAC; an ROS scavenger) or Z-VAD-FMK (a caspase inhibitor) significantly attenuated TMEM230-induced apoptosis in both cultured cells and primary neurons. Our results indicated that TMEM230 mediated a PARP1-linked apoptotic cell death pathway. These findings not only provide the novel insight into the biological roles of TMEM230 in the PARP1-linked pathway but also provide a TMEM230-induced cell death mechanism underlying PD pathogenesis.

Prediction of disease-associated mutations in the transmembrane regions of proteins with known 3D structure.

Being able to assess the phenotypic effects of mutations is a much required capability in precision medicine. However, most of the currently available structure-based methods actually predict stability changes caused by mutations rather than their pathogenic potential. There are also no dedicated methods to predict damaging mutations specifically in transmembrane proteins. In this study we developed and applied a machine-learning approach to discriminate between disease-associated and benign point mutations in the transmembrane regions of proteins with known 3D structure. The method, called BorodaTM (BOosted RegressiOn trees for Disease-Associated mutations in TransMembrane proteins), was trained on sequence-, structure-, and energy-derived descriptors. When compared with the state-of-the-art methods, BorodaTM is superior in classifying point mutations in transmembrane regions. Using BorodaTM we have conducted a large-scale mutation analysis in the transmembrane regions of human proteins with known 3D structures. For each protein we generated structural models for all point mutations by replacing each residue to 19 possible residue types. We classified ~1.8 millions point mutations as benign or diseased-associated and made all predictions available as a Web-server at https://www.iitm.ac.in/bioinfo/MutHTP/boroda.php.

Statistical analysis of disease-causing and neutral mutations in human membrane proteins.

Mutations in transmembrane proteins (TMPs) have diverse effects on their structure and functions, which may lead to various diseases. In this present study, we have investigated variations in human membrane proteins and found that negatively charged to positively charged/polar and nonpolar to nonpolar changes are dominant in disease-causing and neutral mutations, respectively. Further, we analyzed the top 10 preferred mutations in 14 different disease classes and found that each class has at least two Arg mutations. Moreover, in cardiovascular diseases and congenital disorders of metabolism, Cys mutations occur more frequently in single-pass proteins, whereas Arg and nonpolar residues are more frequently substituted in multi-pass membrane proteins. The immune system diseases are enriched in C → R and C → Y mutations in inside and outside regions. On the other hand, in the membrane region, E → K and R → Q mutations are prevalent. The comparison of mutations in topologically similar regions of globular and membrane proteins showed that Ser and Thr mutations cause deleterious effects in membrane regions, whereas Cys and charged residues, Asp and Arg are prevalent in the buried regions of globular proteins. Our comprehensive analysis of disease-associated mutations in transmembrane proteins will be useful for developing prediction tools.

Systems Genetics Analysis of Arrhythmogenic Cardiomyopathy Induced by p.S368L Mutation in Transmembrane Protein 43

BackgroundA missense mutation, p.S358L, in the transmembrane protein 43 (TMEM43/LUMA) gene causes arrhythmogenic right ventricular cardiomyopathy type 5 (ARVC5). ARVC5 is characterized by sudden cardiac death and fibro‐fatty replacement of the myocardium, particularly, in the right ventricle (RV). It has been hypothesized that the TMEM43 mutation impacts function via Ppar signaling, major regulators of adipogenesis. Little is known regarding the mechanistic basis of the S358L mutation‐induced effects.ObjectivesThe aim of this study is to discover novel pathways and gene networks associated with the clinical phenotype elicited by the S358L mutation in the Tmem43 gene using a systems genetics approach.MethodsThe mutant Tmem43S358L mouse was created using the conventional knock‐in approach. BXD strains were derived from a cross between C57BL/6J (B6) and DBA/2J (D2) mice. Total RNAs were extracted from heart of mutant Tmem43S358L mice and 40 BXD strains, and hybridized to Affymetrix Mouse Gene 2.0ST arrays. Differential expression of gene was analyzed using two‐way ANOVA and t‐test. Genetic correlation, functional enrichment, and co‐expression network were performed using GeneNetwork (http://genenetwork.org) and Webgestalt (http://www.webgestalt.org) online tools, respectively.ResultsWe found highly significant expression of Tmem43 hearts among the BXD strains with broad variability in expression levels suggesting an important role for Tmem43 in the heart. A co‐expression Tmem43‐gene network consisting of 25 highly associated genes was created and includes many cardiomyopathy‐related genes, including Tmem43, Ppargc1a and Jup, that are highly significantly connected with Tmem43 and each other (r>0.5, p< 0.00075). A significant negative correlation (p<0.01) between Tmem43 expression and heart mass and heart rate was also noted. Levels of high‐density lipoprotein (HDL) in plasma in BXD mice was significantly correlated with Tmem43 on a normal or high‐fat diet. Microarray analysis of the myocardium revealed significant downregulation of Ppargc1a and Pparg (p<0.0001) in Tmem43S358L mice hearts compared to WT littermates. Decrease in Ppargc1a was significantly related with the Tmem43 decrease in RV, suggesting direct inhibition of Ppargc1a by S358L‐Tmem43 mutation.ConclusionsWe conclude that Tmem43 is essential for normal cardiac function. The Tmem43‐S358L mutation causes alterations in gene expression of numerous cardiomyopathy‐associated genes, including Jup, Ppargc1a and Pparg, resulting in the development of ARVC5.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Chronic interstitial lung disease in children.

Children's interstitial lung diseases (chILD) are increasingly recognised and contain many lung developmental and genetic disorders not yet identified in adult pneumology. Worldwide, several registers have been established. The Australasian Registry Network for Orphan Lung Disease (ARNOLD) has identified problems in estimating rare disease prevalence; focusing on chILD in immunocompetent patients, a period prevalence of 1.5 cases per million children and a mortality rate of 7% were determined. The chILD-EU register highlighted the workload to be covered per patient included and provided protocols for diagnosis and initial treatment, similar to the United States chILD network. Whereas case reports may be useful for young physicians to practise writing articles, cohorts of patients can catapult progress, as demonstrated by recent studies on persistent tachypnoea of infancy, hypersensitivity pneumonitis in children and interstitial lung disease related to interferonopathies from mutations in transmembrane protein 173. Translational research has linked heterozygous mutations in the ABCA3 transporter to an increased risk of interstitial lung diseases, not only in neonates, but also in older children and adults. For surfactant dysfunction disorders in infancy and early childhood, lung transplantation was reported to be as successful as in adult patients. Mutual potentiation of paediatric and adult pneumologists is mandatory in this rapidly extending field for successful future development.This brief review highlights publications in the field of paediatric interstitial lung disease as reviewed during the Clinical Year in Review session presented at the 2017 European Respiratory Society (ERS) Annual Congress in Milan, Italy. It was commissioned by the ERS and critically presents progress made as well as drawbacks.

Lack of TMEM230 mutations in patients with familial and sporadic Parkinson's disease in a Taiwanese population.

Mutations in transmembrane protein 230 (TMEM230) have recently been reported to be associated with Parkinson's disease (PD) in a North American population. A highly prevalent mutation, c.550_552delTAGinsCCCGGG (p.*184ProGlyext*5) was found in 3.1% of Chinese familial PD patients. However, subsequent studies failed to replicate these findings in different populations. Our objective was to confirm the role of this gene in a large number of PD patients and controls in a Taiwanese population. Among 1,672 participants, we sequenced all coding exons and exon-intron boundary junctions of the TMEM230 gene in 180 probands with familial PD. We also genotyped the potential pathogenic variants identified and the previously reported mutations (p.Arg141Leu, p.Tyr92Cys, p.*184Trpext*5, and p.*184ProGlyext*5) in an additional cohort of 500 patients with sporadic PD, and 992 age and gender-matched neurologically normal control subjects. We did not find any of the previously reported mutations, but we observed one novel missense exonic variant, c.G68A (p.Arg23Gln), in one patient with familial PD, and two patients with sporadic PD in a heterozygous state. However, subsequent analysis of this variant in 992 controls did not find any significant associations between p.Arg23Gln and the risk of PD (0.44% vs. 0.30%, p = 0.22). Our findings suggest that genetic variants of TMEM230 do not play a major role in PD in our Taiwanese population. Further experimental studies are warranted to confirm the pathogenicity of this gene in PD disease process.

Subfertility in bulls carrying a nonsense mutation in transmembrane protein 95 is due to failure to interact with the oocyte vestments.

In a recent genome-wide association study, 40 Fleckvieh bulls with exceptionally poor fertility were found to be homozygous for a nonsense mutation in the transmembrane protein 95 (TMEM95) encoding gene. Ejaculates from these individuals exhibited normal sperm concentration, morphology, viability, and motility. However, only 1.7% of inseminations resulted in pregnancies. The aim of this study was to examine the effect of this mutation in TMEM95 on bovine sperm function in vitro. Sperm from homozygous (mt/mt) males had lower in vitro fertility than sperm from wild-type (wt/wt) or heterozygous (wt/mt) bulls (P <0.01). In addition, early embryo division was affected in the mt/mt group (P <0.01). This translated into a lower (P <0.01) blastocyst rate at day 8. Fluorescent staining revealed that TMEM95 is lost after the acrosome reaction. This led us to hypothesize that TMEM95 might be involved in events that lead to sperm-oocyte interaction. After fertilization, a lower number (P <0.01) of sperm from mt/mt bulls bound to the zona pellucida (ZP). Sperm from mt/mt bulls were also less able to penetrate oocytes with no ZP (P<0.01). However, when sperm from these animals were injected into mouse oocytes, they could decondense as successfully as sperm from wt/wt bulls. No differences between genotypes were observed in the ability of sperm to retain motility in an ex vivo oviduct, or in the percentage of sperm exhibiting markers for capacitation and acrosomal reaction. These results suggest that fertilization failure in mt/mt bulls is due to the inability of their sperm to interact with the oocyte vestments.

Disease-associated mutations identify a novel region in human STING necessary for the control of type I interferon signaling

Gain-of-function mutations intransmembrane protein 173 (TMEM173) encoding stimulator of interferon genes (STING) underlie a recently described type I interferonopathy called STING-associated vasculopathy with onset in infancy (SAVI). We sought to define the molecular and cellular pathology relating to 3 individuals variably exhibiting the core features of the SAVI phenotype including systemic inflammation, destructive skin lesions, and interstitial lung disease. Genetic analysis, conformational studies, in vitro assays and ex vivo flow-cytometry were performed. Molecular and in vitro data demonstrate that the pathology in these patients is due to amino acid substitutions at positions 206, 281, and 284 of the human STING protein. These mutations confer cGAMP-independent constitutive activation of type I interferon signaling through TBK1 (TANK-binding kinase), independent from the alternative STING pathway triggered by membrane fusion of enveloped RNA viruses. This constitutive activation was abrogated by ex vivo treatment with the janus kinase 1/2 inhibitor ruxolitinib. Structural analysis indicates that the 3 disease-associated mutations at positions 206, 281, and 284 of the STING protein define a novel cluster of amino acids with functional importance in the regulation of type I interferon signaling.

Characterization of Disease-Associated Mutations in Human Transmembrane Proteins.

Transmembrane protein coding genes are commonly associated with human diseases. We characterized disease causing mutations and natural polymorphisms in transmembrane proteins by mapping missense genetic variations from the UniProt database on the transmembrane protein topology listed in the Human Transmembrane Proteome database. We found characteristic differences in the spectrum of amino acid changes within transmembrane regions: in the case of disease associated mutations the non-polar to non-polar and non-polar to charged amino acid changes are equally frequent. In contrast, in the case of natural polymorphisms non-polar to charged amino acid changes are rare while non-polar to non-polar changes are common. The majority of disease associated mutations result in glycine to arginine and leucine to proline substitutions. Mutations to positively charged amino acids are more common in the center of the lipid bilayer, where they cause more severe structural and functional anomalies. Our analysis contributes to the better understanding of the effect of disease associated mutations in transmembrane proteins, which can help prioritize genetic variations in personal genomic investigations.

Spinocerebellar ataxia type 21 exists in the Chinese Han population.

Recently, mutations in transmembrane protein 240 (TMEM240) were identified as the cause of spinocerebellar ataxia type 21 (SCA21) in several French families. Clinically, SCA21 is characterized as an early-onset, slowly progressive cerebellar syndrome typically associated with cognitive impairment. To date, molecular screening of SCA21 has not been reported among patients of other ethnic origins or in other areas. Here we used Sanger sequencing to detect mutations in exons of TMEM240 in 340 unrelated probands with spinocerebellar ataxia for whom commonly known causative mutations have been excluded (96 probands of autosomal dominant spinocerebellar ataxia families and 244 patients with sporadic spinocerebellar ataxia). As a result, a de novo missense mutation (c.509C > T/p.P170L) was identified in one sporadic SCA patient. The condition manifested as early-onset (30 years old), slowly progressive cerebellar ataxia accompanied by mild early evidenced mental retardation, mild frontal behavior disorders and intentional hand tremors. Although rare, a SCA21 case was identified and described in mainland China, thus broadening the ethnic distribution of SCA21 beyond French families.

NET37, a Nuclear Envelope Transmembrane Protein with Glycosidase Homology, Is Involved in Myoblast Differentiation

The nuclear lamina and its associated proteins are important for nuclear structure and chromatin organization and also have been implicated in the regulation of cell signaling and gene expression. In this study we demonstrate that the lamina-associated nuclear envelope transmembrane protein NET37 is required for myogenic differentiation of C2C12 cells. NET37, a member of glycosidase family 31, is highly expressed in mouse skeletal muscle and is strongly up-regulated during C2C12 differentiation. By protease mapping we show that its glycosidase homology domain is located in the lumen of the nuclear envelope/endoplasmic reticulum. When NET37 is depleted from proliferating myoblasts by RNAi, myogenic differentiation is significantly impaired, and there is a concomitant delay in up-regulation of the late myogenic transcription factor myogenin. We expressed silencing-resistant NET37 mutated at a conserved residue in the glycosidase domain and found that this predicted catalytically inactive protein is unable to support myogenesis in cells depleted of wild type NET37. Therefore, the enzymatic function of NET37 appears to be important for myogenic differentiation. C2C12 cells depleted of NET37 have reduced activation of Akt after shifting to differentiation medium and are defective in insulin like growth factor-II (IGF-II) secretion, an autocrine/paracrine factor involved in Akt activation. We also observed that pro-IGF-II co-immunoprecipitates with NET37. Based on our results, we propose that NET37 has a role in IGF-II maturation in the secretory pathway during myoblast differentiation. The localization of NET37 at the nuclear envelope raises the possibility that it may coordinate myogenic events between the nuclear interior and the endoplasmic reticulum lumen via transmembrane communication.

Moving towards therapies for Juvenile Batten disease?

The neuronal ceroid lipofuscinoses (NCLs or Batten disease) are a group of at least nine autosomal recessively inherited monogenetic storage disorders. Because there are no effective therapies available, all forms of NCL invariably prove fatal after a prolonged period of disability. Indeed, for the forms of NCL that are the result of mutations in transmembrane proteins, the therapeutic outlook remains uniformly bleak. This includes juvenile NCL (JNCL); the most prevalent form of Batten disease that is the result of mutations in the CLN3 gene. Characterizing Cln3 deficient mice is now revealing important clues about the pathogenesis of JNCL. This includes evidence for elevated levels of glutamate within the JNCL CNS and cell type selective sensitivity to α-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) type glutamate receptor overactivation. These findings raise the possibility that AMPA-receptor blockade may potentially be beneficial in JNCL. This possibility has now begun to be tested in Cln3 mutant mice using a single intraperitoneal injection of the non-competitive AMPA antagonist EGIS-8332 by Kovács and Pearce [Kovács, A.D., Pearce, D.A., 2008. Attenuation of AMPA receptor activity improves motor skills in a mouse model of juvenile Batten disease. Exp. Neurol. 209, 288–291.]. Although a positive effect of upon motor coordination deficits in this mouse model of JNCL is reported in this acute study, it remains unclear whether EGIS-8332 provides any lasting benefit or effects upon other aspects of their disease phenotype. Although supplying the first evidence for any form of improvement in a disease-relevant phenotype in Cln3 mutant mice, more detailed studies will be needed to determine whether these preliminary findings will translate into a successful therapy for either murine or human JNCL.