المؤلفون: Jérôme Gilleron, Abderrahman Chafik, Sandra Lacas-Gervais, Jean-François Tanti, Mireille Cormont

المصدر: Cellular & Molecular Biology Letters, Vol 29, Iss 1, Pp 1-21 (2024)

مصطلحات موضوعية: Endosome, Trans Golgi network, Cation-independent mannose-6-phosphate receptor, VPS52, Cytology, QH573-671

الوصف: Abstract Background The trafficking of cargoes from endosomes to the trans-Golgi network requires numerous sequential and coordinated steps. Cargoes are sorted into endosomal-derived carriers that are transported, tethered, and fused to the trans-Golgi network. The tethering step requires several complexes, including the Golgi-associated retrograde protein complex, whose localization at the trans-Golgi network is determined by the activity of small GTPases of the Arl and Rab family. However, how the Golgi-associated retrograde protein complex recognizes the endosome-derived carriers that will fuse with the trans-Golgi network is still unknown. Methods We studied the retrograde trafficking to the trans-Golgi network by using fluorescent cargoes in cells overexpressing Rab4b or after Rab4b knocked-down by small interfering RNA in combination with the downregulation of subunits of the Golgi-associated retrograde protein complex. We used immunofluorescence and image processing (Super Resolution Radial Fluctuation and 3D reconstruction) as well as biochemical approaches to characterize the consequences of these interventions on cargo carriers trafficking. Results We reported that the VPS52 subunit of the Golgi-associated retrograde protein complex is an effector of Rab4b. We found that overexpression of wild type or active Rab4b increased early endosomal to trans-Golgi network retrograde trafficking of the cation-independent mannose-6-phosphate receptor in a Golgi-associated retrograde protein complex-dependent manner. Conversely, overexpression of an inactive Rab4b or Rab4b knockdown attenuated this trafficking. In the absence of Rab4b, the internalized cation-independent mannose 6 phosphate receptor did not have access to VPS52-labeled structures that look like endosomal subdomains and/or endosome-derived carriers, and whose subcellular distribution is Rab4b-independent. Consequently, the cation-independent mannose-6-phosphate receptor was blocked in early endosomes and no longer had access to the trans-Golgi network. Conclusion Our results support that Rab4b, by controlling the sorting of the cation-independent mannose-6-phosphate receptor towards VPS52 microdomains, confers a directional specificity for cargo carriers en route to the trans-Golgi network. Given the importance of the endocytic recycling in cell homeostasis, disruption of the Rab4b/Golgi-associated retrograde protein complex-dependent step could have serious consequences in pathologies.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/1689-1392Test

الوصول الحر: https://doaj.org/article/5ad83691a217409291345c0fa586ceecTest

المؤلفون: Alexander Giaramita, Nathan Wise, Jon Vincelette, Rolando De Angelis, Brett E. Crawford, Sherry Bullens, Nicole Galicia, Melanie J. Lo, Jeremy L. Van Vleet, Sushmita M Roy, Gouri Yogalingam, Amanda R. Luu, Linley Mangini, Alessandra d'Azzo, Vishal Agrawal, Glenn Pacheco, Sylvia Fong, Jonathan H. LeBowitz, Terri Christianson, Hio Wong, Roger Lawrence, Britta Handyside, Harry J Sterling, Stuart Bunting, Chuck Hague, Joseph C Chen

المصدر: The Journal of Biological Chemistry

مصطلحات موضوعية: 0301 basic medicine, safety, medicine.medical_specialty, GM1 gangliosidosis, Biochemistry, enzyme replacement therapy (ERT), 03 medical and health sciences, Mice, Internal medicine, Lysosome, biophysics, Lysosomal storage disease, medicine, Animals, cation-independent mannose-6-phosphate receptor, Enzyme Replacement Therapy, unfolded protein response (UPR), Molecular Biology, Ganglioside, Gangliosidosis, GM1, 030102 biochemistry & molecular biology, Chemistry, Chinese hamster ovary cell, Endoplasmic reticulum, neurodegeneration, toxicity, Enzyme replacement therapy, Cell Biology, medicine.disease, beta-Galactosidase, gene therapy, 030104 developmental biology, medicine.anatomical_structure, Endocrinology, GLB1, lysosomal storage disease, Unfolded protein response, lysosome, endoplasmic reticulum stress

الوصف: Autosomal recessive mutations in the galactosidase β1 (GLB1) gene cause lysosomal β-gal deficiency, resulting in accumulation of galactose-containing substrates and onset of the progressive and fatal neurodegenerative lysosomal storage disease, GM1 gangliosidosis. Here, an enzyme replacement therapy (ERT) approach in fibroblasts from GM1 gangliosidosis patients with recombinant human β-gal (rhβ-gal) produced in Chinese hamster ovary cells enabled direct and precise rhβ-gal delivery to acidified lysosomes. A single, low dose (3 nm) of rhβ-gal was sufficient for normalizing β-gal activity and mediating substrate clearance for several weeks. We found that rhβ-gal uptake by the fibroblasts is dose-dependent and saturable and can be competitively inhibited by mannose 6-phosphate, suggesting cation-independent, mannose 6-phosphate receptor-mediated endocytosis from the cell surface. A single intracerebroventricularly (ICV) administered dose of rhβ-gal (100 μg) resulted in broad bilateral biodistribution of rhβ-gal to critical regions of pathology in a mouse model of GM1 gangliosidosis. Weekly ICV dosing of rhβ-gal for 8 weeks substantially reduced brain levels of ganglioside and oligosaccharide substrates and reversed well-established secondary neuropathology. Of note, unlike with the ERT approach, chronic lentivirus-mediated GLB1 overexpression in the GM1 gangliosidosis patient fibroblasts caused accumulation of a prelysosomal pool of β-gal, resulting in activation of the unfolded protein response and endoplasmic reticulum stress. This outcome was unsurprising in light of our in vitro biophysical findings for rhβ-gal, which include pH-dependent and concentration-dependent stability and dynamic self-association. Collectively, our results highlight that ICV-ERT is an effective therapeutic intervention for managing GM1 gangliosidosis potentially more safely than with gene therapy approaches.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::1842a37d379f3461db62d067305f34f9Test
http://europepmc.org/articles/PMC7521651Test

المؤلفون: Ni-Chung Lee, Wen Hui Tsai, Yin-Hsiu Chien, Pao Chin Chiu, Dwight D. Koeberl, Chaw Liang Chang, Fuu Jen Tsai, Wuh-Liang Hwu

المصدر: Molecular Genetics and Metabolism Reports
Molecular Genetics and Metabolism Reports, Vol 11, Iss C, Pp 31-35 (2017)

مصطلحات موضوعية: 0301 basic medicine, CRIM, cross-reactive immunologic material, 030105 genetics & heredity, NBS, newborn screening, SCT, 4-stair climb test, 0302 clinical medicine, Endocrinology, 6-Min walk test, GMFM, Gross Motor Function Measure, lcsh:QH301-705.5, CI-MPR, cation-independent mannose-6-phosphate receptor, lcsh:R5-920, GAA, acid alpha-glucosidase, Pompe disease, Enzyme replacement therapy, ERT, enzyme replacement therapy, Glc4, glucose tetrasaccharide, Cohort, Ambulatory, medicine.symptom, lcsh:Medicine (General), LOPD, late-onset Pompe disease, AE, adverse event, medicine.drug, Research Paper, medicine.medical_specialty, 6MWT, 6-minute walk test, Nausea, IOPD, infantile-onset Pompe disease, 03 medical and health sciences, Drug withdrawal, Internal medicine, Genetics, medicine, Albuterol, Creatine kinase, Adverse effect, Molecular Biology, rhGAA, recombinant human GAA, business.industry, medicine.disease, Surgery, lcsh:Biology (General), 4-Stair climb test, Adjunctive treatment, Salbutamol, business, MRI, magnetic resonance imaging, 030217 neurology & neurosurgery, CK, creatine kinase

الوصف: Background Early initiation of enzyme replacement therapy (ERT) with recombinant human acid alpha-glucosidase is an effective treatment for patients with infantile-onset Pompe disease (IOPD) but cannot prevent a slow progression of myopathy. Albuterol has been shown to be helpful in adult patients with Pompe disease, and therefore, we administered an open-label adjunctive therapy with albuterol in IOPD patients undergoing ERT. Methods Fourteen patients, aged 2 to 12 years, were enrolled in this study; all of them had a disease onset before 12 months of life, and 13 of them were ambulatory because of early initiation of ERT. All patients received albuterol (also referred to as salbutamol) 12 mg daily for 26 weeks. The outcome measurements included a 6-minute walk test, four-stair climb test (SCT), the standing/walking/running/jumping domains of Gross Motor Function Measure-88, speech quality, serum creatine kinase, and urinary glucose tetrasaccharide. Outcome and safety measurements were evaluated at baseline, and at 1, 3, and 6 months (26 weeks) after entering the trial. Results After a period of 26 weeks, among the 12 patients who were able to complete the SCT, the median time needed decreased by 22% (p = 0.034). Other parameters inconsistently improved in a variety of individuals. Eleven adverse events, including nausea, urinary frequency, and tachycardia, were potentially related to the study drug, but all were mild and disappeared after a brief drug withdrawal. One patient was actively withdrawn from the trial because of poor compliance. Conclusions The results of our study suggest that albuterol showed a good safety profile as an adjunctive treatment in our IOPD cohort, although the benefits are limited.
Highlights • Albuterol may benefit some children with infantile-onset Pompe disease. • Albuterol is safe as an adjunctive treatment to enzyme replacement therapy in children with Pompe disease. • Adverse events associated with albuterol are usually manageable.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::e64331e0852e79ccf95645ab2e3e4a34Test
http://europepmc.org/articles/PMC5406275Test

المؤلفون: Xinmin Fan, Zhangang Xiao, Chunman Li, Bilian Xiong, Ou Sha, Desheng Lu, Tzi Bun Ng, Jing Shen, Meiqi Zeng, Huju Chi, Guangyi Jin

المصدر: Oncotarget

مصطلحات موضوعية: 0301 basic medicine, Male, Cell Membrane Permeability, trichosanthin, medicine.medical_treatment, tumor cells, Apoptosis, Trichosanthin, Granzymes, Receptor, IGF Type 2, 03 medical and health sciences, Mice, 0302 clinical medicine, Cancer immunotherapy, Downregulation and upregulation, Cell Line, Tumor, medicine, Cytotoxic T cell, Animals, Humans, Protein Interaction Domains and Motifs, cation-independent mannose-6-phosphate receptor, Amino Acid Sequence, Receptor, biology, business.industry, granzyme B (GrzB), Cell Membrane, Immunotherapy, Xenograft Model Antitumor Assays, Granzyme B, Disease Models, Animal, 030104 developmental biology, Oncology, Granzyme, 030220 oncology & carcinogenesis, Immunology, Cancer research, biology.protein, immunotherapy, business, Protein Binding, Research Paper

الوصف: // Chunman Li 1 , Meiqi Zeng 1 , Huju Chi 1 , Jing Shen 2 , Tzi-Bun Ng 3 , Guangyi Jin 4 , Desheng Lu 4 , Xinmin Fan 4 , Bilian Xiong 1 , Zhangang Xiao 2 , Ou Sha 1 1 Department of Anatomy, Histology and Developmental Biology, School of Basic Medical Sciences, Shenzhen University Health Science Centre, Shenzhen, Guangdong, China 2 Laboratory of Molecular Pharmacology, Department of Pharmacology, School of Pharmacy, Southwest Medical University, Luzhou, Sichuan, China 3 School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong, Hong Kong, China 4 School of Basic Medical Sciences, Shenzhen University Health Science Centre, Shenzhen, Guangdong, China Correspondence to: Ou Sha, email: shaou@szu.edu.cn Zhangang Xiao, email: xzg555898@hotmail.com Keywords: trichosanthin, granzyme B (GrzB), cation-independent mannose-6-phosphate receptor, tumor cells, immunotherapy Received: December 23, 2016 Accepted: February 08, 2017 Published: February 19, 2017 ABSTRACT Trichosanthin is a plant toxin belonging to the family of ribosome-inactivating proteins. It has various biological and pharmacological activities, including anti-tumor and immunoregulatory effects. In this study, we explored the potential medicinal applications of trichosanthin in cancer immunotherapy. We found that trichosanthin and cation-independent mannose-6-phosphate receptor competitively bind to the Golgi-localized, γ-ear containing and Arf-binding proteins. It in turn promotes the translocation of cation-independent mannose-6-phosphate receptor from the cytosol to the plasma membrane, which is a receptor of Granzyme B. The upregulation of this receptor on the tumor cell surface increased the cell permeability to Granzyme B, and the latter is one of the major factors of cytotoxic T lymphocyte-mediated tumor cell apoptosis. These results suggest a novel potential application of trichosanthin and shed light on its anti-tumor immunotherapy.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::d70b0eb2428e97e785734e95e0b4ad16Test
https://pubmed.ncbi.nlm.nih.gov/28460437Test

المؤلفون: Shinji Saiki, Soichiro Kakuta, Masato Koike, Masumi Sato, Atsuhito Fuse, Aki Inose, Nobutaka Hattori, Norihiko Furuya

المصدر: FEBS letters. 589(13)

مصطلحات موضوعية: Proteasome Endopeptidase Complex, Retromer, Endosome, Protein subunit, Blotting, Western, Biophysics, SNXs, Vesicular Transport Proteins, Endosomes, Biochemistry, Transport Pathway, Cation independent mannose-6-phosphate receptor, VPS35, Structural Biology, Genetics, Humans, Molecular Biology, Microscopy, Confocal, Proteasome, Chemistry, Ubiquitin, Cell Biology, Transport protein, Cell biology, Retromer complex, Microscopy, Electron, Protein Transport, VPS29, Multiprotein Complexes, Proteolysis, RNA Interference, VPS26, HeLa Cells, trans-Golgi Network

الوصف: Retromer is a complex of proteins that functions in the endosome-to-Golgi retrieval cargo transport pathway. VPS35 works as the central subunit of retromer to recognize the cargos and binds with VPS29 and VPS26 via distinct domains. We show that deficiency of VPS35 or VPS29 accompanies degradation of other subunits, whereas VPS26 deficiency had no effect on VPS29 and VPS35 levels. Although VPS35 forms VPS26–VPS35 and VPS29–VPS35 sub-complexes with similar efficiency in vitro, VPS26–VPS35 was more easily degradable by the ubiquitin–proteasome-system than VPS29–VPS35. These results indicate that VPS29 and VPS35 form a biologically stable sub-complex in vivo.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::707ff1e3ac8dcbe567eef25f0869a98fTest
https://pubmed.ncbi.nlm.nih.gov/25937119Test

المؤلفون: Rajawat, Yogendra Singh

المساهمون: Bossis, Ioannis, Digital Repository at the University of Maryland, University of Maryland (College Park, Md.), Veterinary Medical Science

مصطلحات موضوعية: Veterinary medicine, Cellular biology, Molecular biology, All trans retinoic acid, Autophagosome maturation, Autophagy, Cation-independent Mannose-6-Phosphate Receptor, LC3, Vitamin A

الوصف: Retinoic acids (RAs) have diverse biologic effects and regulate several cellular functions. Here, we investigated the role of RA on autophagy by studying its effects on autophagosome (AUT) maturation, as well as on upstream regulators of autophagosome biogenesis. Our studies, based on the use of pH-sensitive fluorescent reporter markers, suggest that RA promotes AUT acidification and maturation. By using competitive inhibitors and specific agonists, we demonstrated that this effect is not mediated by the classic Retinoic Acid Receptor (RAR) and Retinoid X Receptors (RXR). RA did not affect the protein expression levels of upstream regulators of autophagy, such as Beclin-1, phospho-mTOR, and phospho-Akt1, but induced redistribution of both endogenous cation-independent mannose-6-phosphate receptor CI-MPR and transiently transfected GFP and RFP full-length CI-MPR fusion proteins from the trans-Golgi region to acidified AUT structures. Those structures were found to be amphisomes (acidified AUTs) and not autophagolysosomes. The critical role of CI-MPR in AUT maturation was further demonstrated by siRNA-mediated silencing of endogenous CI-MPR. Transient CI-MPR knockdown resulted in remarkable accumulation of nonacidified AUTs, a process that could not be reversed with RA.These results suggest that RA induces AUT acidification and maturation by regulating CI-MPR subcellular location, a process critical in the cellular autophagic mechanism.

وصف الملف: application/pdf

العلاقة: http://hdl.handle.net/1903/14646Test

الإتاحة: http://hdl.handle.net/1903/14646Test

مستخلص: Retinoic acids (RAs) have diverse biologic effects and regulate several cellular functions. Here, we investigated the role of RA on autophagy by studying its effects on autophagosome (AUT) maturation, as well as on upstream regulators of autophagosome biogenesis. Our studies, based on the use of pH-sensitive fluorescent reporter markers, suggest that RA promotes AUT acidification and maturation. By using competitive inhibitors and specific agonists, we demonstrated that this effect is not mediated by the classic Retinoic Acid Receptor (RAR) and Retinoid X Receptors (RXR). RA did not affect the protein expression levels of upstream regulators of autophagy, such as Beclin-1, phospho-mTOR, and phospho-Akt1, but induced redistribution of both endogenous cation-independent mannose-6-phosphate receptor CI-MPR and transiently transfected GFP and RFP full-length CI-MPR fusion proteins from the trans-Golgi region to acidified AUT structures. Those structures were found to be amphisomes (acidified AUTs) and not autophagolysosomes. The critical role of CI-MPR in AUT maturation was further demonstrated by siRNA-mediated silencing of endogenous CI-MPR. Transient CI-MPR knockdown resulted in remarkable accumulation of nonacidified AUTs, a process that could not be reversed with RA.These results suggest that RA induces AUT acidification and maturation by regulating CI-MPR subcellular location, a process critical in the cellular autophagic mechanism.

مصطلحات الفهرس: Veterinary medicine, Cellular biology, Molecular biology, All trans retinoic acid, Autophagosome maturation, Autophagy, Cation-independent Mannose-6-Phosphate Receptor, LC3, Vitamin A, Dissertation

URL: http://hdl.handle.net/1903/14646Test