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Effects of miR-219/miR-338 on microglia and astrocyte behaviors and astrocyte-oligodendrocyte precursor cell interactions

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Neural Regeneration Research 2020年第4期15卷 739-747页

作者： Lan Huong Nguyen William Ong Kai Wang Mingfeng Wang Dean Nizetic Sing Yian Chew School of Chemical and Biomedical Engineering Nanyang Technological UniversitySingaporeSingapore Lee Kong Chian School of Medicine Nanyang Technological UniversitySingaporeSingapore Blizard Institute Barts&The London School of MedicineQueen Mary University of LondonLondonUK NUS Synthetic Biology for Clinical and Technological Innovation(SynCTI) Life Sciences InstituteNational University of SingaporeSingapore

MiR-219 and miR-338(miR-219/miR-338)are oligodendrocyte-specific *** overexpression of these miRs in oligodendrocyte precursor cells promotes their differentiation and maturation into oligodendrocytes,which may enhance axonal remyelination after nerve injuries in the central nervous system(CNS).As such,the delivery of miR-219/miR-338 to the CNS to promote oligodendrocyte precursor cell differentiation,maturation and myelination could be a promising approach for nerve ***,nerve injuries in the CNS also involve other cell types,such as microglia and ***,we investigated the effects of miR-219/miR-338 treatment on microglia and astrocytes in vitro and in *** found that miR-219/miR-338 diminished microglial expression of pro-inflammatory cytokines and suppressed astrocyte *** addition,we showed that miR-219/miR-338 enhanced oligodendrocyte precursor cell differentiation and maturation in a scratch assay paradigm that re-created a nerve injury condition in ***,our results suggest miR-219/miR-338 as a promising treatment for axonal remyelination in the CNS following nerve *** experimental procedures were approved by the Institutional Animal Care and Use Committee(IACUC),Nanyang Technological University(approval No.A0309 and A0333)on April 27,2016 and October 8,2016.

关键词： central nervous system electrospinning gene silencing glia hydrogel myelination nanofibers oligodendroglial polycaprolactone spinal cord injury

RNAi mechanisms in Huntington’s disease therapy:siRNA versus shRNA

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Translational Neurodegeneration 2017年第1期6卷 300-309页

作者： Sebastian Aguiar Bram van der Gaag Francesco Albert Bosco Cortese Molecular Neuroscience Laboratory Swammerdam Institute for Life Sciences(SILS-CNS)University of AmsterdamAmsterdamNetherlands Fulbright Program US Department of State(IIE)New York CityNYUSA Department of Neurobiology Care Sciences and SocietyKarolinska InstituteStockholmSweden Biogerontology Research Foundation(BGRF) OxfordUK Department of Biomedical and Molecular Sciences Queen’s University School of MedicineQueen’s UniversityKingstonCanada

Huntington’s Disease(HD)is a genetically dominant trinucleotide repeat disorder resulting from CAG repeats within the Huntingtin(HTT)gene exceeding a normal range(>36 CAGs).Symptoms of the disease manifest in middle age and include chorea,dystonia,and cognitive *** latency from diagnosis to death is 20 *** are currently no disease-modifying therapies available to HD *** is a potentially curative therapy for HD.A popular line of research employs siRNA or antisense oligonucleotides(ASO)to knock down mutant Huntingtin mRNA(mHTT).Unfortunately,this modality requires repeated dosing,commonly exhibit off target effects(OTEs),and exert renal and hepatic *** contrast,a single AAV-mediated short-hairpin RNA(shRNA)dose can last years with low *** addition,we highlight research indicating that shRNA elicits fewer OTEs than siRNA when tested *** this promise,shRNA therapy has been held back by difficulties controlling expression(oversaturating cells with toxic levels of RNA construct).In this review,we compare RNAi modalities for HD and propose novel methods of optimizing shRNA expression and on-target fidelity.

关键词： RNAi shRNA siRNA Huntington’s disease Off-target effects Huntingtin silencing

Poly(ADP-ribose)polymerase family member 14 promotes functional recovery after spinal cord injury through regulating microglia M1/M2 polarization via STAT1/6 pathway

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Neural Regeneration Research 2023年第8期18卷 1809-1817页

作者： Ai-Hua Xu Yang Yang Yang Shao Man-Yu Jiang Yong-Xin Sun Department of Rehabilitation Medicine The First Hospital of China Medical UniversityShenyangLiaoning ProvinceChina

Poly(ADP-ribose)polymerase family member 14(PARP14),which is an intracellular mono(ADP-ribosyl)transferase,has been reported to promote post-stroke functional recovery,but its role in spinal cord injury(SCI)remains *** investigate this,a T10 spinal cord contusion model was established in C57BL/6 mice,and immediately after the injury PARP14 shRNA-carrying lentivirus was injected 1 mm from the injury site to silence PARP14 *** found that PARP14 was up-regulated in the injured spinal cord and that lentivirus-mediated downregulation of PARP14 aggravated functional impairment after injury,accompanied by obvious neuronal apoptosis,severe neuroinflammation,and slight bone ***,PARP14 levels were elevated in microglia after SCI,PARP14 knockdown activated microglia in the spinal cord and promoted a shift from M2-polarized microglia(anti-inflammatory phenotype)to M1-polarized microglia(pro-inflammatory phenotype)that may have been mediated by the signal transducers and activators of transcription(STAT)1/6 ***,microglia M1 and M2 polarization were induced in vitro using lipopolysaccharide/interferon-γand interleukin-4,*** results showed that PARP14 knockdown promoted microglia M1 polarization,accompanied by activation of the STAT1 *** addition,PARP14 overexpression made microglia more prone to M2 polarization and further activated the STAT6 *** conclusion,these findings suggest that PARP14 may improve functional recovery after SCI by regulating the phenotypic transformation of microglia via the STAT1/6 pathway.

关键词： apoptosis M1 polarization M2 polarization microglia neuroinflammation PARP14 silencing spinal cord injury STAT1 pathway STAT6 pathway

The role of water channel proteins and nitric oxide signaling in rice seed germination

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Cell Research 2007年第7期17卷 638-649页

作者： Sang-Soo Kwak Korea Research Institute of Bioscience and Biotechnology Oun-dong 52 Yusong Taejon 305-333 South Korea

Previous studies have demonstrated the possible role of several aquaporins in seed germination. But systematic inves-tigation of the role of aquaporin family members in this process is lacking. Here, the developmental regulation of plasmamembrane intrinsic protein (PIP) expression throughout germination and post-germination processes in rice embryoswas analyzed. The expression patterns of the PIPs suggest these aquaporins play different roles in seed germination andseedling growth. Partial silencing of the water channel genes, OsPIP1;1 and OsPIP1;3, reduced seed germination whileover-expression of OsPIP1;3 promoted seed germination under water-stress conditions. Moreover, spatial expressionanalysis indicates that OsPIP1;3 is expressed predominantly in embryo during seed germination. Our data also revealedthat the nitric oxide (NO) donors, sodium nitroprusside (SNP) and S-nitrosoglutathione (GSNO), promoted seed germi-nation; furthermore, the NO scavenger, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, inhibitedgermination and reduced the stimulative effects of SNP and GSNO on rice germination. Exogenous NO stimulated thetranscription of OsPIP1;1, OsPIP1;2, OsPIP1;3 and OsPIP2;8 in germinating seeds. These results suggest that waterchannels play an important role in seed germination, acting, at least partly, in response to the NO signaling pathway.

关键词： germination nitric oxide Oryza sativa over-expression silencing water channel

Efficient Self-Assembled DNA Nanoparticles through Rolling Circle Amplification for siRNA Delivery in vitro

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Chinese Journal of Chemistry 2019年第6期37卷 588-592页

作者： Qian Yao Yuqi Chen Fan Wu1 Fan Wu Chaoxing Liu Tingting Hong Wei Li Yi Chen Xiang Zhou College of Chemistry and Molecular Sciences Key Laboratory of Biomedical Polymers of Ministry of EducationHubei Province Key Laboratory of Allergy and ImmunologyWuhan UniversityWuhanHubei 430072China

Effective and low toxicity delivery of siRNA is of great importance for clinical gene therapy. Herein, self-assembled DNA nanoparticles (NPs) based on rolling circle amplification (RCA) with a small interfering RNA (siRNA) payload were successfully developed as a facile and efficient siRNA delivery strategy. This intracellular gene silencing strategy exhibits various advantages including low toxicity, high efficiency, and good stability. The synthesized DNA NPs serve as siRNA carriers, protecting the siRNA against nuclease degradation. We demonstrate that the obtained self-assembled siRNA/NP/PEI system can successfully deliver enhanced green fluorescent protein (EGFP)-siRNA into HeLa cells, realizing the same EGFP knockdown efficiency with less toxicity as that of commercial Lipofectamine 2000.

关键词： siRNA delivery self-assembly RCA DNA nanopartides gene silencing

Design, mechanism, delivery and therapeutics of canonical and Dicer-substrate siRNA

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Asian Journal of Pharmaceutical Sciences 2019年第5期14卷 497-510页

作者： Maria Abdul Ghafoor Raja Haliza Katas Muhammad Wahab Amjad Department of Pharmaceutics Faculty of PharmacyNorthern Border UniversityRafha73211Saudi Arabia Centre for Drug Delivery Research Faculty of PharmacyUniversiti Kebangsaan MalaysiaKuala Lumpur50300Malaysia

Upon the discovery of RNA interference(RNAi),canonical small interfering RNA(si RNA) has been recognized to trigger sequence-specific gene silencing. Despite the benefits of si RNAs as potential new drugs,there are obstacles still to be overcome,including off-target effects and immune stimulation. More recently,Dicer substrate si RNA(Dsi RNA) has been introduced as an alternative to si RNA. Similarly,it also is proving to be potent and target-specific,while rendering less immune stimulation. Dsi RNA is 25–30 nucleotides in length,and is further cleaved and processed by the Dicer enzyme. As with si RNA,it is crucial to design and develop a stable,safe,and efficient system for the delivery of Dsi RNA into the cytoplasm of targeted cells. Several polymeric nanoparticle systems have been well established to load Dsi RNA for in vitro and in vivo delivery,thereby overcoming a major hurdle in the therapeutic uses of Dsi RNA. The present review focuses on a comparison of si RNA and Dsi RNA on the basis of their design,mechanism,in vitro and in vivo delivery,and therapeutics.

关键词： RNA interference Drug delivery system Polymeric nanoparticles Gene silencing Gene carrier Non-viral vector

RhoC,vascular endothelial growth factor and microvascular density in esophageal squamous cell carcinoma

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World Journal of Gastroenterology 2015年第3期21卷 905-912页

作者： Zhi-Hua Zhao Yan Tian Jian-Pin Yang Jun Zhou Kui-Sheng Chen Department of Pathology the First Affiliated Hospital of Zhengzhou University

AIM:To investigate the expression of Ras homolog(Rho)C,vascular endothelial growth factor(VEGF) and CD105 in esophageal squamous cell ***:Semi-quantitative reverse transcriptase polymerase chain reaction,in situ hybridization and immunohistochemical streptavidin-biotin- peroxidase methods were used to detect expression of Rho C m RNA and protein,and VEGF protein in 62 cases with esophageal squamous cell carcinoma,31 cases with adjacent atypical hyperplastic tissues,and 62 cases with normal esophageal ***105 antibody labeling was used to measure microvascular *** levels were compared according to clinicopathologic and patient ***:Expression of Rho C m RNA showed a positive correlation with the protein level in esophageal squamous cell carcinoma,as well as with VEGF protein *** C m RNA expression was mainly located within the cytoplasm of the tumor cells,appearing as blue to purple particles by in situ *** differences in Rho C m RNA expression in esophageal squamous cell carcinoma,adjacent atypical hyperplasia and normal esophageal mucosa were significant(P < 0.05).The relative expression of Rho C m RNA in cancer tissues with lymph node metastasis was significantly higher than in the tissues without lymph node metastasis(P < 0.05).VEGF protein expression was consistent with microvascular density(t = 25.52,P < 0.05).Positive expression of VEGF protein in esophageal squamous cell carcinoma of different histologic gradings did not differ *** expression of VEGF protein in carcinoma tissues with deep infiltration was significantly higher than in tissues with only superficial infiltration(P < 0.05).The positive expression of VEGF protein in cancer tissues with lymph node metastasis was significantly higher than in the tissues without lymph node metastasis(P < 0.05).CONCLUSION:Rho C protein may upregulate VEGF expression,thereby promoting tumor *** C m RNA and protein expre

关键词： Esophageal squamous cell carcinoma Gene silencing

Influence of DNA methylation on transgene expression

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Chinese Science Bulletin 2001年第15期46卷 1300-1304页

作者： Xugang Li Zhen Zhu Dejiang Feng Tuanjie Chang Xiang Liu Chinese Acad Sci Inst Genet Beijing 100101 Peoples R China

DNA methylation plays an important role in gene expression in eukaryote. But DNA methylation of transgene usually leads to target gene silencing in plant genetic engineering. In this research, reporter gene β-glu-curonidase (GUS) gene (uidA) was introduced into tobaccos via Agrobacterium-mediated transformation method, and the foreign uidA gene became inactive in some transgenic tobaccos. No mRNA of uidA was detected in these plants by Northern blotting analysis, and DNA methylation of promoter region was found. The results indicated that gene silencing might be caused by DNA methylation of promoter.

关键词： DNA methylation gene silencing transgene GUS tran-scriptional product.

Molecular Adaptation of Peanut Metabolic Pathways to Wide Variations of Mineral Ion Composition and Concentration

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American Journal of Plant Sciences 2012年第1期3卷 33-50页

作者： Godson O. Osuji Tassine K. Brown Sanique M. South Justin C. Duncan Dwiesha Johnson Shanique Hyllam 不详

Plant evolution, nutritional genomics, and mineral nutrition have been well documented but no studies have focused on the molecular adaptation of crop metabolism to wide variations of mineral ion composition and concentration. Diversification of peanut species from primary centers of domestication in South America depended on metabolic adaptation to the mineral ion conditions of the newer habitats. Understanding the diversification molecular biology of peanut metabolic pathways will permit the synthesis of the best mineral ion combinations for doubling CO2 assimilation. Valencia and Virginia cultivars belong to different subspecies of the tetraploid Arachis hypogaea. They were planted in the absence and presence of up to 99 mM (equivalent to 166 moles per hectare) of different mineral ions. Molecular properties of the primary metabolic pathways were studied by Northern analyses using Valencia GDH-synthesized RNAs as probes for Virginia mRNA and GDH-synthesized RNAs. Messenger RNAs are silenced by homologous RNAs synthesized by GDH. Peanut cellulose was analyzed by gravimetry;and fatty acids by HPLC. Complementary DNA probes made from Valencia GDH-synthesized RNAs hybridized perfectly to Virginia mRNAs and GDH-synthesized RNAs. Wide variations in mineral ion compositions and concentrations induced the GDHs of Valencia and Virginia to synthesize RNAs that differentially down-regulated the mRNAs encoding phosphate translocator, granule-bound starch synthase, phosphoglucomutase, glucosyltransferase, acetyl CoA carboxylase, nitrate reductase, and NADH-glutamate synthase so that the percent weights of oil (41.53 ± 8.75) and cellulose (30.29 ± 3.12) were similar in the control and mineral-treated peanuts. Therefore, RNA sequences that defined the molecular adaptation of mRNAs encoding the enzymes of primary metabolism were the same in the varietal types of A. hypogaea, in agreement with genetic data suggesting that tetraploid Arachis evolved relatively recently from the wi

关键词： Glutamate Dehydrogenase-Synthesized RNA mRNA silencing Phosphate Translocator Acetyl CoA Carboxylase Cellulosic Biomass Northern Analysis HPLC Fatty Acids