Disruption of lysosomal nutrient sensing scaffold contributes to pathogenesis of a fatal neurodegenerative lysosomal storage disease.

The ceroid lipofuscinosis neuronal 1 (CLN1) disease, formerly called infantile neuronal ceroid lipofuscinosis, is a fatal hereditary neurodegenerative lysosomal storage disorder. This disease is caused by loss-of-function mutations in the CLN1 gene, encoding palmitoyl-protein thioesterase-1 (PPT1). PPT1 catalyzes depalmitoylation of S-palmitoylated proteins for degradation and clearance by lysosomal hydrolases. Numerous proteins, especially in the brain, require dynamic S-palmitoylation (palmitoylation-depalmitoylation cycles) for endosomal trafficking to their destination. While 23 palmitoyl-acyl transferases in the mammalian genome catalyze S-palmitoylation, depalmitoylation is catalyzed by thioesterases such as PPT1. Despite these discoveries, the pathogenic mechanism of CLN1 disease has remained elusive. Here, we report that in the brain of Cln1-/- mice, which mimic CLN1 disease, the mechanistic target of rapamycin complex-1 (mTORC1) kinase is hyperactivated. The activation of mTORC1 by nutrients requires its anchorage to lysosomal limiting membrane by Rag GTPases and Ragulator complex. These proteins form the lysosomal nutrient sensing scaffold to which mTORC1 must attach to activate. We found that in Cln1-/- mice, two constituent proteins of the Ragulator complex (vacuolar (H+)-ATPase and Lamtor1) require dynamic S-palmitoylation for endosomal trafficking to the lysosomal limiting membrane. Intriguingly, Ppt1 deficiency in Cln1-/- mice misrouted these proteins to the plasma membrane disrupting the lysosomal nutrient sensing scaffold. Despite this defect, mTORC1 was hyperactivated via the IGF1/PI3K/Akt-signaling pathway, which suppressed autophagy contributing to neuropathology. Importantly, pharmacological inhibition of PI3K/Akt suppressed mTORC1 activation, restored autophagy, and ameliorated neurodegeneration in Cln1-/- mice. Our findings reveal a previously unrecognized role of Cln1/Ppt1 in regulating mTORC1 activation and suggest that IGF1/PI3K/Akt may be a targetable pathway for CLN1 disease.

Inhibition of Wnt/ß-catenin signaling reduces renal fibrosis in murine glycogen storage disease type Ia.

Glycogen storage disease type Ia (GSD-Ia) is caused by a deficiency in the enzyme glucose-6-phosphatase-α (G6Pase-α or G6PC) that is expressed primarily in the gluconeogenic organs, namely liver, kidney cortex, and intestine. Renal G6Pase-α deficiency in GSD-Ia is characterized by impaired gluconeogenesis, nephromegaly due to elevated glycogen accumulation, and nephropathy caused, in part, by renal fibrosis, mediated by activation of the renin-angiotensin system (RAS). The Wnt/ß-catenin signaling regulates the expression of a variety of downstream mediators implicated in renal fibrosis, including multiple genes in the RAS. Sustained activation of Wnt/ß-catenin signaling is associated with the development and progression of renal fibrotic lesions that can lead to chronic kidney disease. In this study, we examined the molecular mechanism underlying GSD-Ia nephropathy. Damage to the kidney proximal tubules is known to trigger acute kidney injury (AKI) that can, in turn, activate Wnt/ß-catenin signaling. We show that GSD-Ia mice have AKI that leads to activation of the Wnt/ß-catenin/RAS axis. Renal fibrosis was demonstrated by increased renal levels of Snail1, α-smooth muscle actin (α-SMA), and extracellular matrix proteins, including collagen-Iα1 and collagen-IV. Treating GSD-Ia mice with a CBP/ß-catenin inhibitor, ICG-001, significantly decreased nuclear translocated active ß-catenin and reduced renal levels of renin, Snail1, α-SMA, and collagen-IV. The results suggest that inhibition of Wnt/ß-catenin signaling may be a promising therapeutic strategy for GSD-Ia nephropathy.

Anti-ulcerogenic potential of anthocyanin-loaded chitosan nanoparticles against alcohol-HCl induced gastric ulcer in rats.

Chitosan is more prominent in food applications due to its versatile properties. Anthocyanins have gained much research attention due to their multifaceted role in preventing various lifestyle ailments. Encapsulated anthocyanin- loaded chitosan nanoparticles (ACNPs) were prepared by conventional ionotropic gelation method. In the present study, the gastro-protective effect of encapsulated ACNPs was evaluated against absolute ethanol-hydrochloric acid (HCl-Ethanol mixture) induced gastric ulcer in male Wistar rats. The histopathology and microscopic scoring of ulcer data of stomach tissue sections revealed that oral administration of encapsulated ACNPs group can alleviate inflammation of induced-gastric ulcer. Further, the expression of anti-inflammatory cytokines (Interleukin 4, IL-4) and suppression of pro-inflammatory cytokines (Interferon gamma, IFN-γ) confirm the cytoprotective effect of encapsulated ACNPs against HCl-Ethanol induced necrotic damage to mucosal membrane. The results of the present study indicate that the gastro protective action of encapsulated ACNPs ascribable to at least in parts to its anti-inflammatory property.[Formula: see text].

Sulfated polysaccharide ascophyllan from Padina tetrastromatica enhances healing of burn wounds by ameliorating inflammatory responses and oxidative damage.

Sulfated polysaccharide ascophyllan from marine brown algae has been identified to have burn wound healing properties. Thus, we examined the effects of ascophyllan fraction (AF3) on the inflammatory response and oxidative damage in burn wounds. Full-thickness burn wounds in rats were then treated twice per day with topical AF3 ointment (5%), while control groups were treated with 10% povidone-iodine (positive control) and petroleum jelly-based ointment (negative control). The activity of cyclooxygenase-2 and myeloperoxidase and levels of C-reactive protein, nitric oxide, and proinflammatory cytokines (tumor necrosis factor-α, interleukin-6, and interleukin-1ß) were observed to have significantly decreased in peripheral blood mononuclear cells, serum, and wound tissue of the group treated with AF3 ointment on day 8 after wounding. The expression of inducible nitric oxide synthase, endothelial nitric oxide synthase, and vascular endothelial growth factor at the mRNA level was determined to be upregulated in the wound tissue of the AF3 ointment-treated group. After treatment with AF3 ointment, the antioxidant enzyme activity and level of reduced glutathione were upregulated, whereas the content of thiobarbituric acid reactive substances decreased. Treatment of burn wounds using 5% AF3 ointment decreases oxidative damage associated with inflammation deceptively via inhibition of inflammatory enzymes, regulation of proinflammatory cytokines, upregulation of angiogenesis, and activity of antioxidant enzymes.