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https://www.readbyqxmd.com/read/28944845/efficient-inhibition-of-duck-hepatitis-b-virus-dna-by-the-crispr-cas9-system
#1
Qingfen Zheng, Li Bai, Sujun Zheng, Mei Liu, Jinyan Zhang, Ting Wang, Zhongwei Xu, Yu Chen, Jiansheng Li, Zhongping Duan
Current therapeutic strategies cannot eradicate hepatitis B virus covalently closed circular DNA (HBV cccDNA), which accounts for the persistence of HBV infection. Very recently, the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR‑associated protein 9 (Cas9) system has been used as an efficient and powerful tool for viral genome editing. Given that the primary duck hepatocyte (PDH) infected with duck hepatitis B virus (DHBV) has been widely used to study human HBV infection in vitro, the present study aimed to demonstrate the targeted inhibition of DHBV DNA, especially cccDNA, by the CRISPR/Cas9 system using this model...
September 19, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28915572/down-regulation-of-ntcp-expression-by-cyclin-d1-in-hepatitis-b-virus-related-hepatocellular-carcinoma-has-clinical-significance
#2
Jingting Kang, Jie Wang, Jin Cheng, Zhiliang Cao, Ran Chen, Huiyu Li, Shuang Liu, Xiangmei Chen, Jianhua Sui, Fengmin Lu
The sodium-dependent taurocholate cotransporter polypeptide (NTCP) has been identified as a liver specific functional receptor for the hepatitis B virus (HBV). Previous studies indicated that the expression of NTCP may be associated with the proliferation status of hepatocytes. However, the involvement of NTCP in hepatocellular carcinoma (HCC) cells proliferation remains unclear. In this study, we confirmed that NTCP was down-regulated in HCC tumor tissues compared with that in the adjacent non-tumor tissues (P < 0...
August 22, 2017: Oncotarget
https://www.readbyqxmd.com/read/28887167/identification-of-slug-and-sox7-as-transcriptional-repressors-binding-to-the-hepatitis-b-virus-core-promoter
#3
Hui Ling Ko, Tze Hau Lam, Huijin Ng, Jiaying Toh, Liang Wei Wang, Ee Chee Ren
BACKGROUND & AIMS: The Hepatitis B Virus (HBV) is carried in many non-liver cell types but does not actively replicate in them. We investigated the possibility that these cells possess HBVCP transcription inhibitory mechanisms specifically absent in liver cells, which together with other liver-specific mechanisms such as NTCP-mediated entry, enable liver cells to effectively produce HBV. METHODS: Liver and non-liver cell lines were screened for their capacity to activate the HBVCP and synthesize pgRNA...
September 5, 2017: Journal of Hepatology
https://www.readbyqxmd.com/read/28870671/relationship-between-serum-hbv-rna-levels-and-intrahepatic-viral-as-well-as-histologic-activity-markers-in-entecavir-treated-patients
#4
Jing Wang, Yiqi Yu, Guojun Li, Chuan Shen, Zhefeng Meng, Jianming Zheng, Yanhong Jia, Shaolong Chen, Xiao Zhang, MengQi Zhu, Jiangjiang Zheng, Zhangzhang Song, Jing Wu, Lingyun Shao, Peiyu Qian, Xiaona Mao, Xuanyi Wang, Yuxian Huang, Caiyan Zhao, Jiming Zhang, Chao Qiu, Wenhong Zhang
BACKGROUND: In diagnostics, serum hepatitis B virus (HBV)-RNA levels are valuable when the HBV-DNA load in circulation is effectively suppressed by nucleos(t)ide analogue (NUC) therapy. This study aimed to determine the intrahepatic viral replication activity reflected in serum HBV-RNA and whether HBV-RNA contributes to liver histological changes in NUC-treated patients. METHODS: A cross-sectional set of serum and liver biopsy samples was obtained from entecavir-treated patients with undetectable levels of serum HBV-DNA...
September 1, 2017: Journal of Hepatology
https://www.readbyqxmd.com/read/28839466/the-grna-mirna-grna-ternary-cassette-combining-crispr-cas9-with-rnai-approach-strongly-inhibits-hepatitis-b-virus-replication
#5
Jie Wang, Ran Chen, Ruiyang Zhang, Shanlong Ding, Tianying Zhang, Quan Yuan, Guiwen Guan, Xiangmei Chen, Ting Zhang, Hui Zhuang, Frederick Nunes, Timothy Block, Shuang Liu, Zhongping Duan, Ningshao Xia, Zhongwei Xu, Fengmin Lu
The CRISPR/Cas9 system is a novel genome editing technology which has been successfully used to inhibit HBV replication. Here, we described a novel gRNA-microRNA (miRNA)-gRNA ternary cassette driven by a single U6 promoter. With an anti-HBV pri-miR31 mimic integrated between two HBV-specific gRNAs, both gRNAs could be separated from the long transcript of gRNA-miR-HBV-gRNA ternary cassette through Drosha/DGCR8 processing. The results showed that the gRNA-miR-HBV-gRNA ternary cassette could efficiently express two gRNAs and miR-HBV...
2017: Theranostics
https://www.readbyqxmd.com/read/28833361/epigenetic-regulation-of-hepatitis-b-virus-covalently-closed-circular-dna-implications-for-epigenetic-therapy-against-chronic-hepatitis-b
#6
REVIEW
Xupeng Hong, Elena S Kim, Haitao Guo
Hepatitis B virus (HBV) infection represents a significant public health burden worldwide. Although current therapeutics manage to control the disease progression, lifelong treatment and surveillance are required because drug resistance develops during treatment and reactivations frequently occur following medication cessation. Thus, the occurrence of hepatocellular carcinoma (HCC) is decreased but not eliminated. One major reason for the treatment failure is the inability to eradicate or inactivate the viral covalently closed circular DNA (cccDNA) which is a stable episomal form of viral genome decorated with host histones and non-histone proteins...
August 18, 2017: Hepatology: Official Journal of the American Association for the Study of Liver Diseases
https://www.readbyqxmd.com/read/28827084/inhibitory-effects-of-metachromin-a-on-hepatitis-b-virus-production-via-impairment-of-the-viral-promoter-activity
#7
Atsuya Yamashita, Mayumi Tamaki, Hirotake Kasai, Tomohisa Tanaka, Teruhime Otoguro, Akihide Ryo, Shinya Maekawa, Nobuyuki Enomoto, Nicole J de Voogd, Junichi Tanaka, Kohji Moriishi
The currently available antiviral agents for chronic infection with hepatitis B virus (HBV) are pegylated interferon-α and nucleoside/nucleotide analogues, although it has been difficult to completely eliminate covalently closed circular DNA (cccDNA) from patients. To identify an antiviral compound targeting HBV core promoter, 15 terpenes originating from marine organisms were screened using a cell line expressing firefly luciferase under the control of the HBV core promoter. Metachromin A, which is a merosesquiterpene isolated from the marine sponge Dactylospongia metachromia, inhibited the viral promoter activity at the highest level among the tested compounds, and suppressed HBV production with an EC50 value of 0...
August 4, 2017: Antiviral Research
https://www.readbyqxmd.com/read/28802063/hepatitis-b-virus-pregenomic-rna-in-hepatocellular-carcinoma-a-nosological-and-prognostic-determinant
#8
Boris Halgand, Christophe Desterke, Lise Rivière, Guillaume Fallot, Mylène Sebagh, Julien Calderaro, Paulette Bioulac-Sage, Christine Neuveut, Marie-Annick Buendia, Didier Samuel, Cyrille Féray
Hepatitis B virus (HBV) is a major cause of hepatocellular carcinoma (HCC). However, very little is known about the replication of HBV in HCC tissues. PATIENTS AND METHODS: We analysed viral and cellular parameters in HCC (T) and non-tumor liver (NT) samples from 99 HBsAg-positive, virologically suppressed patients treated by tumour resection or liver transplantation. We examined total HBV DNA and RNA as well as covalently closed circular DNA (cccDNA) and pregenomic RNA (pgRNA), which are considered as markers of active HBV replication...
August 12, 2017: Hepatology: Official Journal of the American Association for the Study of Liver Diseases
https://www.readbyqxmd.com/read/28774415/new-treatments-to-reach-functional-cure-virological-approaches
#9
REVIEW
David Durantel
Current therapies of chronic hepatitis B (CHB) remain limited to pegylated-interferon-alpha (pegIFN-α) or any of the five approved nucleos(t)ide analogues (NA). If viral suppression can be achieved in the majority of patients with the high-barrier-to-resistance new-generation of NA, i.e. entecavir and tenofovir, HBsAg loss is achieved by PEG-IFN-α and/or NA in only 10% of patients, after a 5-year follow-up. Attempts to improve the response by administering two different NA or a combination of NA and PEG-IFN-α have not provided a dramatic increase in the rate of "functional cure"...
June 2017: Best Practice & Research. Clinical Gastroenterology
https://www.readbyqxmd.com/read/28774407/the-virological-aspects-of-hepatitis-b
#10
REVIEW
Zina S Valaydon, Stephen A Locarnini
Human hepatitis B virus (HBV) is a hepatotropic virus that is responsible for a significant burden of disease, causing liver disease and hepatocellular carcinoma. It is a small DNA virus with a replication strategy that is similar to that of a retrovirus. HBV is prone to mutagenesis and under the influence of diverse selection pressures, has evolved into a pool of quasispecies, genotypes and mutants, which confers a significant survival advantage. The genome is small, circular, and compact but has a complex replication strategy...
June 2017: Best Practice & Research. Clinical Gastroenterology
https://www.readbyqxmd.com/read/28750917/predictive-role-of-serum-hbsag-and-hbcrag-kinetics-in-patients-with-hbeag-negative-chronic-hepatitis-b-receiving-pegylated-interferon-based-therapy
#11
Natthaya Chuaypen, Nawarat Posuwan, Salyavit Chittmittraprap, Nattiya Hirankarn, Sombat Treeprasertsuk, Yasuhito Tanaka, Noboru Shinkai, Yong Poovorawan, Pisit Tangkijvanich
OBJECTIVES: To investigate the role of serum hepatitis B core-related antigen (HBcrAg) kinetics in predicting long-term outcome of pegylated interferon (PEG-IFN)-based therapy in patients with HBeAg-negative chronic hepatitis B (CHB). METHODS: 121 Thai patients with HBeAg-negative CHB recruited from a previous randomized trial of 48-week PEG-IFN alone or combined with entecavir were enrolled. HBsAg and HBcrAg levels were serially examined. Paired biopsies at baseline and post-treatment were assessed for intrahepatic cccDNA...
July 24, 2017: Clinical Microbiology and Infection
https://www.readbyqxmd.com/read/28749559/recombinant-cccdna-of-hepatitis-b-virus-induces-long-term-viral-persistence-with-chronic-hepatitis-in-a-mouse-model
#12
Gaiyun Li, Yuanfei Zhu, Dianhui Shao, Hao Chang, Xiaoming Zhang, Dongming Zhou, Yueqiu Gao, Ke Lan, Qiang Deng
Covalently closed circular (ccc) DNA of hepatitis B virus (HBV) is critical for viral persistence in vivo. We recently reported a technique involving recombinant (r) cccDNA of HBV by site-specific DNA recombination. Using hydrodynamic injection, rcccDNA induces a temporarily prolonged HBV anigenemia in immunocompetent mice, similar to acute resolving HBV infection. In this study, we simulated the pathophysiological impact of chronic hepatitis by which to reproduce (r)cccDNA persistence in mouse models. We showed that rcccDNA achieved long-lasting persistence in the presence of a compromised immune response or when transcriptional activity was repressed...
July 27, 2017: Hepatology: Official Journal of the American Association for the Study of Liver Diseases
https://www.readbyqxmd.com/read/28747369/serum-hbv-dna-rna-and-hbsag-which-correlated-better-to-intrahepatic-covalently-closed-circular-dna-before-and-after-nucleos-t-ide-analogue-treatment
#13
Yuhua Gao, Yutang Li, Qinghua Meng, Zhanqing Zhang, Ping Zhao, Qinghua Shang, Yue Li, Mingze Su, Tong Li, Xueen Liu, Hui Zhuang
The study was to investigate whether serum HBV RNA is a strong surrogate marker for intrahepatic HBV covalently closed circular DNA (cccDNA) when compared with serum HBV DNA, hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) in HBeAg-positive chronic hepatitis B (CHB) patients. Serum HBV RNA, HBV DNA, HBsAg, HBeAg and intrahepatic cccDNA were quantitatively detected at baseline (n=82) and 96 weeks (n=62) after nucleos(t)ide analogues (NUC) treatment in HBeAg-positive CHB patients. Then the correlations between serum HBV RNA, HBV DNA, HBsAg, HBeAg and intrahepatic cccDNA were statistically analyzed...
July 26, 2017: Journal of Clinical Microbiology
https://www.readbyqxmd.com/read/28725013/baseline-value-of-intrahepatic-hbv-dna-over-cccdna-predicts-patient-s-response-to-interferon-therapy
#14
Di Mu, Fang-Chao Yuan, Yu Chen, Xiao-Yan Jiang, Liang Yan, Ling-Yu Jiang, Jian-Ping Gong, Da-Zhi Zhang, Hong Ren, Yong Liao
Methodology for accurate quantification of intra-hepatic cccDNA has long been a technical challenge, yet it is highly desired in the clinic. Here, we developed a sensitive method for quantification of intrahepatic cccDNA in liver biopsies from patients, which allowed to predict patient's response to interferon therapy at baseline. Twenty-five patients with HBeAg+ CHB were recruited and liver biopsies were obtained at baseline and 1-year after interferon treatment, respectively. Both intrahepatic cccDNA and HBV DNA were absolutely quantified by a droplet digital PCR amplification system...
July 19, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28709658/ngago-gdna-system-efficiently-suppresses-hepatitis-b-virus-replication-through-accelerating-decay-of-pregenomic-rna
#15
Zhuanchang Wu, Siyu Tan, Leiqi Xu, Lifen Gao, Haizhen Zhu, Chunhong Ma, Xiaohong Liang
Covalently closed circular DNA (cccDNA) in the hepatocytes nucleus is responsible for persistent infection of Hepatitis B virus (HBV). Current antiviral therapy drugs nucleos(t)ide analogs or interferon fail to eradicate HBV cccDNA. Genome editing technique provides an effective approach for HBV treatment through targeting viral cccDNA. Natronobacterium gregoryi Argonaute (NgAgo)-guide DNA (gDNA) system with powerful genome editing prompts us to explore its application in inhibiting HBV replication. Preliminary function verification indicated that NgAgo/EGFP-gDNA obviously inhibited EGFP expression...
July 12, 2017: Antiviral Research
https://www.readbyqxmd.com/read/28709657/detection-of-the-hepatitis-b-virus-hbv-covalently-closed-circular-dna-cccdna-in-mice-transduced-with-a-recombinant-aav-hbv-vector
#16
Julie Lucifora, Anna Salvetti, Xavier Marniquet, Laurent Mailly, Barbara Testoni, Floriane Fusil, Aurore Inchauspé, Maud Michelet, Marie-Louise Michel, Massimo Levrero, Pierre Cortez, Thomas F Baumert, François-Loic Cosset, Cécile Challier, Fabien Zoulim, David Durantel
Hepatitis B Virus (HBV) persists in infected hepatocytes as an episomal covalently-closed-circular DNA mini-chromosome, called cccDNA. As the main nuclear transcription template, HBV cccDNA is a key replication intermediate in the viral life cycle. Little is known about the mechanisms involved in its formation, maintenance and fate under antiviral therapies. This is mainly due to the lack of small immune-competent animal models able to recapitulate the entire HBV replication cycle, including formation of HBV cccDNA...
July 11, 2017: Antiviral Research
https://www.readbyqxmd.com/read/28709006/compartmental-hbv-evolution-and-replication-in-liver-and-extrahepatic-sites-after-nucleos-tide-analogue-therapy-in-chronic-hepatitis-b-carriers
#17
Shan Gao, Zhong-Ping Duan, Yu Chen, Frank van der Meer, Samuel S Lee, Carla Osiowy, Guido van Marle, Carla S Coffin
BACKGROUND: Hepatitis B virus (HBV) variants are associated with nucleos/tide analogue (NA) response and liver disease but it is unknown whether NA influences extrahepatic HBV persistence. OBJECTIVES: To investigate HBV replication and genetic evolution in hepatic and extrahepatic sites of chronic hepatitis B (CHB) before and after NA therapy. STUDY DESIGN: A total of 13 paired plasma, peripheral blood mononuclear cells (PBMC), were collected from chronic HBV carriers at baseline and after a median 53 weeks NA therapy as well as liver biopsy (N=7 baseline, N=5 follow-up)...
September 2017: Journal of Clinical Virology: the Official Publication of the Pan American Society for Clinical Virology
https://www.readbyqxmd.com/read/28696237/persistent-loss-of-hepatitis-b-virus-markers-in-serum-without-cellular-immunity-by-combination-of-peginterferon-and-entecavir-therapy-in-humanized-mice
#18
Takuro Uchida, Michio Imamura, C Nelson Hayes, Nobuhiko Hiraga, Hiromi Kan, Masataka Tsuge, Hiromi Abe-Chayama, Yizhou Zhang, Grace Naswa Makokha, Hiroshi Aikata, Daiki Miki, Hidenori Ochi, Yuji Ishida, Chise Tateno, Kazuaki Chayama
Nucleot(s)ide analogues and peginterferon (PEG-IFN) treatment are the only approved therapies for chronic hepatitis B virus (HBV) infection. However, complete eradication of the virus, as indicated by persistent loss of hepatitis B surface antigen (HBsAg), is rare among treated patients. This is due to long-term persistence of the HBV genome in infected hepatocytes in the form of covalently closed circular DNA (cccDNA). In this study, we investigated whether administration of a large dose of a nucleoside analogue in combination with PEG-IFN can achieve long-term loss of HBsAg in human hepatocyte chimeric mice...
September 2017: Antimicrobial Agents and Chemotherapy
https://www.readbyqxmd.com/read/28694331/transcriptional-elongation-control-of-hepatitis-b-virus-covalently-closed-circular-dna-transcription-by-super-elongation-complex-and-brd4
#19
Joel Celio Francisco, Qian Dai, Zhuojuan Luo, Yan Wang, Roxanne Hui-Heng Chong, Yee Joo Tan, Wei Xie, Guan-Huei Lee, Chengqi Lin
Chronic hepatitis B virus (HBV) infection can lead to liver cirrhosis and hepatocellular carcinoma. HBV reactivation during or after chemotherapy is a potentially fatal complication for cancer patients with chronic HBV infection. Transcription of HBV is a critical intermediate step of the HBV life cycle. However, factors controlling HBV transcription remain largely unknown. Here, we found that different P-TEFb complexes are involved in the transcription of the HBV viral genome. Both BRD4 and the super elongation complex (SEC) bind to the HBV genome...
October 1, 2017: Molecular and Cellular Biology
https://www.readbyqxmd.com/read/28688280/synthesis-of-sulfamoylbenzamide-derivatives-as-hbv-capsid-assembly-effector
#20
Ozkan Sari, Sebastien Boucle, Bryan D Cox, Tugba Ozturk, Olivia Ollinger Russell, Leda Bassit, Franck Amblard, Raymond F Schinazi
The synthesis of novel series of sulfamoylbenzamides as HBV capsid assembly effector is reported. The structure was divided into five parts which were independently modified as part of our lead optimization. All synthesized compounds were evaluated for their anti-HBV activity and toxicity in human hepatocytes, lymphocytes and other cells. Additionally, we assessed their effect on HBV cccDNA formation in an HBeAg reporter cell-based assay. Among the 27 compounds reported, several analogs exhibited submicromolar activities and significant reduction of HBeAg secretion...
September 29, 2017: European Journal of Medicinal Chemistry
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