Rescue of Advanced Pompe Disease in Mice with Hepatic Expression of Secretable Acid α-Glucosidase.

Pompe disease is a neuromuscular disorder caused by disease-associated variants in the gene encoding for the lysosomal enzyme acid α-glucosidase (GAA), which converts lysosomal glycogen to glucose. We previously reported full rescue of Pompe disease in symptomatic 4-month-old Gaa knockout (Gaa-/-) mice by adeno-associated virus (AAV) vector-mediated liver gene transfer of an engineered secretable form of GAA (secGAA). Here, we showed that hepatic expression of secGAA rescues the phenotype of 4-month-old Gaa-/- mice at vector doses at which the native form of GAA has little to no therapeutic effect. Based on these results, we then treated severely affected 9-month-old Gaa-/- mice with an AAV vector expressing secGAA and followed the animals for 9 months thereafter. AAV-treated Gaa-/- mice showed complete reversal of the Pompe phenotype, with rescue of glycogen accumulation in most tissues, including the central nervous system, and normalization of muscle strength. Transcriptomic profiling of skeletal muscle showed rescue of most altered pathways, including those involved in mitochondrial defects, a finding supported by structural and biochemical analyses, which also showed restoration of lysosomal function. Together, these results provide insight into the reversibility of advanced Pompe disease in the Gaa-/- mouse model via liver gene transfer of secGAA.This work was supported by Genethon, the French Muscular Dystro-phy Association (AFM), and Spark Therapeutics. It was also sup-ported by the European Union’s Research and Innovation Programunder grant agreement number 667751 (to F.M.), the EuropeanResearch Council Consolidator Grant under grant agreement number617432 (to F.M.), and Marie Skłodowska-Curie Actions-IndividualFellowship (MSCA-IF) grant agreement number 797144 (to U.C.)S

Benzotriazole Derivatives as Long Wavelength Photosensitizers for Diaryliodonium Salt Initiators

Two benzotriazole derivative dyes 4,7-bis(2,3-dihydrothieno[3,4-b][1,4]dioxin-5-yl)-2-dodecyl-2H-benzo[1,2,3]triazole, and 2-dodecyl-4,7-bis(4-hexylthiophen-2-yl)-2H-benzo[d][1,2,3]triazole are shown to work as efficient photosensitizers for a dipheny-liodonium salt initiator in cationic photopolymerization of epoxide and vinyl monomers. Substituted thienyl groups are attached to benzotriazole backbone to extend conjugation and enhance electron density of the molecules. Thereby, it was possible to initiate polymerizations at room temperature using long wavelength UV and visible light. The progress of photopolymerizations was monitored using optical pyrometry. The photopolymerization of an epoxide monomer using solar irradiation was also demonstrated. (C) 2010 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 49: 729-733, 201

Is manubrium limited mini-sternotomy an alternative for traditional full sternotomy in cases with massive mediastinal (retrosternal) goiter? A case report

Although most mediastinal goiters (MG) are operable by a cervical approach, some cases require sternotomy. Manubrium limited mini-sternotomy (MLMS) is an alternative method for traditional full sternotomy in subjects with MG. We present the case of a 45-year-old male subject who was operated for a total MG. The nodular mass was conical in shape and extended beyond the aortic arch and tracheal bifurcation. Sternotomy was decided because excision via cervical Kocher incision was not safe. The nodular mass was extracted from the mediastinum easily via MLMS. The postoperative period was uneventful, without any complication, and the patient was discharged from the hospital after two days. He returned to his normal life within one month. This case shows that MLMS is a convenient and easy alternative for traditional full sternotomy in subjects with MG

The effects of L-carnitine administration on energy metabolism in pregnant Halep (Damascus) goats

The aim of this study was to determine the effects of parenteral administration of L-carnitine on some biochemical parameters in Halep (Damascus) goats during the last month of pregnancy. L-carnitine was administrated to goats in group I (n = 13) by subcutaneous injections once a week during the last month of the pregnancy. Physiologic salt solution was administered to goats in group II (n = 12) by the same route during the same period. Differences of glucose concentration between groups were not significant (P > 0.05). Serum beta-hydroxybutyric acid (BHB) concentrations in both groups increased until parturition. However, differences between groups were not significant (P > 0.05). Concentration of serum NEFA (Non Esterified Fatty Acid) in group I was lower compared to group II 2 weeks before parturition (P 0.05). Level of glucose concentration in L-carnitine administered goats with twin kids was higher than the controls with twin kids in the 2(nd) (P < 0.01) and 3(rd) weeks (P < 0.05) before parturition. It was concluded that parenteral administration of L-carnitine might be a protective measure against pregnancy toxemia (ketosis) via increasing serum glucose concentration in goats with twin pregnancy

Mitochondrial retrograde signaling regulates neuronal function

Mitochondria are key regulators of cellular homeostasis, and mitochondrial dysfunction is strongly linked to neurodegenerative diseases, including Alzheimer’s and Parkinson’s. Mitochondria communicate their bioenergetic status to the cell via mitochondrial retrograde signaling. To investigate the role of mitochondrial retrograde signaling in neurons, we induced mitochondrial dysfunction in the Drosophila nervous system. Neuronal mitochondrial dysfunction causes reduced viability, defects in neuronal function, decreased redox potential, and reduced numbers of presynaptic mitochondria and active zones. We find that neuronal mitochondrial dysfunction stimulates a retrograde signaling response that controls the expression of several hundred nuclear genes. We show that the Drosophila hypoxia inducible factor alpha (HIFα) ortholog Similar (Sima) regulates the expression of several of these retrograde genes, suggesting that Sima mediates mitochondrial retrograde signaling. Remarkably, knockdown of Sima restores neuronal function without affecting the primary mitochondrial defect, demonstrating that mitochondrial retrograde signaling is partly responsible for neuronal dysfunction. Sima knockdown also restores function in a Drosophila model of the mitochondrial disease Leigh syndrome and in a Drosophila model of familial Parkinson’s disease. Thus, mitochondrial retrograde signaling regulates neuronal activity and can be manipulated to enhance neuronal function, despite mitochondrial impairment

Gene therapy with secreted acid alpha-glucosidase rescues Pompe disease in a novel mouse model with early-onset spinal cord and respiratory defects.

Pompe disease (PD) is a neuromuscular disorder caused by deficiency of acidalpha-glucosidase (GAA), leading to motor and respiratory dysfunctions. Available Gaa knock-out (KO) mouse models do not accurately mimic PD, particularly its highly impaired respiratory phenotype. Here we developed a new mouse model of PD crossing Gaa KOB6;129 with DBA2/J mice. We subsequently treated Gaa KODBA2/J mice with adeno-associated virus (AAV) vectors expressing a secretable form of GAA (secGAA). Male Gaa KODBA2/J mice present most of the key features of the human disease, including early lethality, severe respiratory impairment, cardiac hypertrophy and muscle weakness. Transcriptome analyses of Gaa KODBA2/J, compared to the parental Gaa KOB6;129 mice, revealed a profoundly impaired gene signature in the spinal cord and a similarly deregulated gene expression in skeletal muscle. Muscle and spinal cord transcriptome changes, biochemical defects, respiratory and muscle function in the Gaa KODBA2/J model were significantly improved upon gene therapy with AAV vectors expressing secGAA. These data show that the genetic background impacts on the severity of respiratory function and neuroglial spinal cord defects in the Gaa KO mouse model of PD. Our findings have implications for PD prognosis and treatment, show novel molecular pathophysiology mechanisms of the disease and provide a unique model to study PD respiratory defects, which majorly affect patients. This work was supported by Genethon, the French Muscular Dystrophy Association (AFM), the European Commission (grant nos. 667751, 617432, and 797144), and Spark Therapeutics.This work was supported by Genethon and the French Muscular Dystrophy Association (AFM, to F.M.). It was also supported by the European Union’s research and innovation program under grant agreement no. 667751 (to F.M.), the European Research Council Consolidator Grant under grant agreement no. 617432 (to F.M.), Marie Skodowska-Curie Actions Individual Fellowship (MSCA-IF) grant agreement no. 797144 (to U.C.), and by Spark Therapeutics under a sponsored research agreement.S

Unkempt Is Negatively Regulated by mTOR and Uncouples Neuronal Differentiation from Growth Control

<div><p>Neuronal differentiation is exquisitely controlled both spatially and temporally during nervous system development. Defects in the spatiotemporal control of neurogenesis cause incorrect formation of neural networks and lead to neurological disorders such as epilepsy and autism. The mTOR kinase integrates signals from mitogens, nutrients and energy levels to regulate growth, autophagy and metabolism. We previously identified the insulin receptor (InR)/mTOR pathway as a critical regulator of the timing of neuronal differentiation in the <i>Drosophila melanogaster</i> eye. Subsequently, this pathway has been shown to play a conserved role in regulating neurogenesis in vertebrates. However, the factors that mediate the neurogenic role of this pathway are completely unknown. To identify downstream effectors of the InR/mTOR pathway we screened transcriptional targets of mTOR for neuronal differentiation phenotypes in photoreceptor neurons. We identified the conserved gene <i>unkempt</i> (<i>unk</i>), which encodes a zinc finger/RING domain containing protein, as a negative regulator of the timing of photoreceptor differentiation. Loss of <i>unk</i> phenocopies InR/mTOR pathway activation and <i>unk</i> acts downstream of this pathway to regulate neurogenesis. In contrast to InR/mTOR signalling, <i>unk</i> does not regulate growth. <i>unk</i> therefore uncouples the role of the InR/mTOR pathway in neurogenesis from its role in growth control. We also identified the gene <i>headcase</i> (<i>hdc</i>) as a second downstream regulator of the InR/mTOR pathway controlling the timing of neurogenesis. Unk forms a complex with Hdc, and Hdc expression is regulated by <i>unk</i> and InR/mTOR signalling. Co-overexpression of <i>unk</i> and <i>hdc</i> completely suppresses the precocious neuronal differentiation phenotype caused by loss of <i>Tsc1</i>. Thus, Unk and Hdc are the first neurogenic components of the InR/mTOR pathway to be identified. Finally, we show that Unkempt-like is expressed in the developing mouse retina and in neural stem/progenitor cells, suggesting that the role of Unk in neurogenesis may be conserved in mammals.</p></div