ライフサイエンスコーパス: cccDNA

1 cccDNA is assembled with cellular histone proteins into

2 he nuclei assayed contained between 1 and 17 cccDNA molecules, with the remaining 10% containing more

3 ssing the virus are known, information about cccDNA formation, stability, and turnover is lacking.

4 Current knowledge about cccDNA stability was largely derived from quantitative a

5 T1 and STAT2 transcription factors to active cccDNA.

6 he relationship between hepatocyte death and cccDNA elimination requires knowing both the amount of h

7 assay for the detection of HBV RNA, DNA, and cccDNA.

8 en-positive (HBsA-positive) and HBV DNA- and cccDNA-positive cells.

9 oplasm and reduction of nuclear DP rcDNA and cccDNA.

10 d the decline in cccDNA, so that the average cccDNA copy number in infected cells dropped during the

11 Because cccDNA does not replicate semiconservatively, it is not

12 DNA by Cas9 and a dramatic reduction in both cccDNA and other parameters of viral gene expression and

13 To fill the gaps in knowledge concerning cccDNA biology, we have developed a fluorescence imaging

14 In contrast, cccDNA declined more slowly, consistent with a half-life

15 gating HBV cccDNA biology and for developing cccDNA-targeting drugs.

16 ded the first insights into the fate of DHBV cccDNA and nuclear HBV DNA under conditions mimicking an

17 e detection of duck hepatitis B virus (DHBV) cccDNA and HBV nuclear DNA in established cell lines.

18 s, including covalently closed circular DNA (cccDNA) and Dane particles, were detected only after ind

19 e same time, covalently closed circular DNA (cccDNA) and viral mRNA levels both declined about two- t

20 es including covalently closed circular DNA (cccDNA) are present.

21 fect initial covalently closed circular DNA (cccDNA) conversion but inhibits the synthesis of progeny

22 coating, and covalently closed circular DNA (cccDNA) formation.

23 s methylated covalently closed circular DNA (cccDNA) in human liver tissue.

24 umulation of covalently closed circular DNA (cccDNA) in nuclei of infected cells.

25 lear pool of covalently closed circular DNA (cccDNA) in the liver.

26 from the HBV covalently closed circular DNA (cccDNA) minichromosome, both in cultured cells in which

27 a long-lived covalently closed circular DNA (cccDNA) molecule, is degraded noncytolytically by agents

28 V) contained covalently closed circular DNA (cccDNA) molecules with deletions and insertions indicati

29 Covalently closed circular DNA (cccDNA) of hepatitis B virus (HBV) is formed by conversi

30 HBV covalently closed circular DNA (cccDNA) plays an essential role in HBV persistence and r

31 ool of viral covalently closed circular DNA (cccDNA) transcriptional template of HBV, which must be e

32 ated DNA and covalently closed circular DNA (cccDNA) were turned over independently of cell division.

33 virus (HBV), covalently closed circular DNA (cccDNA), has been difficult to study in patients with ch

34 omosome, the covalently closed circular DNA (cccDNA), in the nucleus of infected hepatocytes, as well

35 ool of viral covalently closed circular DNA (cccDNA), resulting in a transient elevation of viral rep

36 ctly affect nuclear HBV closed circular DNA (cccDNA), the genomic form that templates viral transcrip

37 Covalently closed circular DNA (cccDNA), the nuclear form of hepatitis B virus (HBV), is

38 ation of new covalently closed circular DNA (cccDNA), the viral transcriptional template.

39 te the viral covalently closed circular DNA (cccDNA), which is a stable episomal form of the viral ge

40 ahepatic WHV covalently closed circular DNA (cccDNA).

41 e stable HBV covalently-closed-circular DNA (cccDNA).

42 ncluding the covalently closed circular DNA (cccDNA).

43 hepadnavirus covalently closed circular DNA (cccDNA).

44 ral genomes [covalently closed circular DNA (cccDNA)] in the nuclei of infected cells.

45 ue to the persistence of viral episomal DNA (cccDNA) in infected cells.

46 It remains unclear if existing cccDNA is eliminated noncytolytically or if hepatocyte d

47 The development of a FISH-based assay for cccDNA tracking provided the first insights into the fat

48 While methods for cccDNA quantification from liver biopsy specimens and ce

49 Each nucleus was assayed by nested PCR for cccDNA and for cellular IFN-alpha genes as an internal c

50 cer and the amount of pgRNA transcribed from cccDNA) were significantly higher in cells expressing wi

51 The mcHBV-GLuc cccDNA model is independent of HBV infection, and will b

52 d to non-HBV minicircle plasmids, mcHBV-GLuc cccDNA showed persistent HBV-GLuc activity and HBx-depen

53 Importantly, the mcHBV-GLuc cccDNA showed resistance to interferons (IFN) treatment,

54 The mcHBV-GLuc cccDNA was easily produced in bacteria, and it formed mi

55 th a Gaussia Luciferase reporter (mcHBV-GLuc cccDNA), which serves as a surrogate to measure cccDNA a

56 cteria, and it formed minichromosomes as HBV cccDNA episome DNA does when it was transfected into hum

57 and nucleic acids and a mismatch between HBV cccDNA, RNA, and expression of the hepatitis B surface a

58 B virus covalently closed circular DNA (HBV cccDNA), the episomal form of the virus that persists de

59 only, host DNA-repair factor involved in HBV cccDNA biogenesis.

60 V entry inhibitors, HBV core inhibitors, HBV cccDNA transcripts RNA interference, HBV cell apoptosis

61 mechanisms, we quantitated intrahepatic HBV cccDNA levels in acutely infected chimpanzees whose viro

62 , and will be valuable for investigating HBV cccDNA biology and for developing cccDNA-targeting drugs

63 We engineered a minicircle HBV cccDNA with a Gaussia Luciferase reporter (mcHBV-GLuc cc

64 We report here a novel HBV cccDNA technology that will meet the need.

65 a, produced by T cells, reduce levels of HBV cccDNA in hepatocytes by inducing deamination and subseq

66 n about the intrahepatic distribution of HBV cccDNA in infected patients, particularly at the single-

67 nistic understanding of the stability of HBV cccDNA in the presence of antiviral therapy and during c

68 -alpha mediates epigenetic repression of HBV cccDNA transcriptional activity, which may assist in the

69 cccDNA is hampered by the lack of robust HBV cccDNA models.

70 ent, indicating its unique similarity to HBV cccDNA that is usually resistant to long-term IFN treatm

71 el it also decreased hepatitis B virus (HBV) cccDNA and woodchuck surface antigen.

72 Most importantly, GLuc illuminates cccDNA as a surrogate of cccDNA activity, providing a ve

73 ted hepatocytes lagged behind the decline in cccDNA, so that the average cccDNA copy number in infect

74 Moreover, the declines in cccDNA appeared to reflect the loss of hepatocytes from

75 ith M and/or S proteins led to a decrease in cccDNA levels, indicating that L contributes to the regu

76 ledge, the first genome-wide maps of PTMs in cccDNA-containing chromatin from de novo infected HepG2

77 gate the role of the cytoplasmic DP rcDNA in cccDNA formation, we demonstrated that rcDNA deproteiniz

78 Administration of IFN-alpha resulted in cccDNA-bound histone hypoacetylation as well as active r

79 t with TPD2's having a physiological role in cccDNA formation, RNAi-mediated TDP2 depletion in human

80 for detecting viral nucleic acids, including cccDNA, with single-cell resolution provides a means for

81 Initially, cccDNA is derived from RC DNA from the infecting virion,

82 A formation, and conversion of DP-rcDNA into cccDNA is a rate-limiting step of cccDNA formation in He

83 the increase of HBV viremia and intrahepatic cccDNA loads was significantly slower than in HBV mono-i

84 Clearances of intrahepatic cccDNA and/or HBsAg are critical endpoints for future an

85 DNA), which serves as a surrogate to measure cccDNA activity.

86 We have therefore measured cccDNA levels in woodchuck hepatocyte cultures following

87 Furthermore, methylated cccDNA was found in tumor and nonneoplastic human liver

88 Here, using a new cccDNA ChIP-Seq approach, we report, to our knowledge, t

89 he generation time of hepatocytes and if new cccDNA formation were effectively blocked.

90 is possible to segregate cells containing no cccDNA.

91 up to 36 days with 3TC reduced the amount of cccDNA in the cultures not more than twofold compared to

92 e and quick method to detect trace amount of cccDNA.

93 in limits the intracellular amplification of cccDNA for duck hepatitis B virus.

94 proteins in regulating the amplification of cccDNA in HBV is not well characterized.

95 argely derived from quantitative analyses of cccDNA levels present in liver samples, and little was k

96 hosen guide RNAs, we demonstrate cleavage of cccDNA by Cas9 and a dramatic reduction in both cccDNA a

97 e infecting virion, but additional copies of cccDNA are derived from newly synthesized RC DNA molecul

98 ect effect on the up to 50 or more copies of cccDNA that maintain the infected state.

99 ma and TNF-alpha each induced deamination of cccDNA and interfered with its stability; their effects

100 Deprivation of cccDNA required activation of nuclear APOBEC3 deaminases

101 In vitro, the desiccation of cccDNA led to loss of supercoiling, aggregation, loss of

102 sing FISH, we determined the distribution of cccDNA under conditions mimicking chronic infections wit

103 The underlying dynamics of cccDNA persistence are unlikely to impact the probabilit

104 te death were responsible for elimination of cccDNA during recovery from transient infections.

105 However, elimination of cccDNA might be facilitated if its half-life were short

106 s one of the major routes for elimination of cccDNA.

107 Finally, an in vitro equivalent of cccDNA showed decreased viral protein production in HepG

108 articular, little is known about the fate of cccDNA during cell division.

109 ples, and little was known about the fate of cccDNA in individual cells.

110 t with the possibility that some fraction of cccDNA was distributed to daughter cells in those infect

111 the formation of two or more generations of cccDNA from linear DNA was observed.

112 DNA target will promote the highest level of cccDNA disruption.

113 ion led to a decrease (>50%) in the level of cccDNA, which inversely correlated with the level of the

114 led to an increase (>6-fold) in the level of cccDNA.

115 ation, the cells accumulate higher levels of cccDNA as well as larger amounts of deproteinized rcDNA

116 The loss of cccDNA was comparable to that expected from the estimate

117 ring antiviral therapy, the rates of loss of cccDNA, infected hepatocytes (1 or more molecules of ccc

118 ter T-cell stimulation prevented the loss of cccDNA.

119 The mechanism of cccDNA formation is unknown, but the release of P protei

120 ce suggests that epigenetic modifications of cccDNA contribute to viral replication and the outcome o

121 infected hepatocytes (1 or more molecules of cccDNA), and replicating DNAs may be quite different.

122 tudies have measured the mean copy number of cccDNA molecules in hepadnaviral-infected cells, the dis

123 a PCR-based assay, we examined the number of cccDNA molecules of the duck hepatitis B virus in single

124 all, the data suggest (i) that the number of cccDNA molecules per cell may fluctuate over time, and (

125 To study the molecular pathway of cccDNA formation and its regulation by viral and cellula

126 The first step reduces the pool of cccDNA molecules noncytolytically, probably by eliminati

127 e, including the mechanism and regulation of cccDNA formation.

128 ssibility that chromatin-based regulation of cccDNA transcription could be a new therapeutic approach

129 ting that L contributes to the regulation of cccDNA.

130 rcDNA into cccDNA is a rate-limiting step of cccDNA formation in HepG2 cells.

131 y, GLuc illuminates cccDNA as a surrogate of cccDNA activity, providing a very sensitive and quick me

132 port demonstrates regulation of synthesis of cccDNA by the envelope proteins of HBV.

133 hepatocytes from the cultures rather than of cccDNA from hepatocytes.

134 espite this multipronged response, traces of cccDNA persist indefinitely in the liver, likely providi

135 A fraction of cccDNAs formed from both type 1 and type 2 linear DNAs a

136 HBV replication with little or no impact on cccDNA, hence lifelong treatment is required in the vast

137 tion products, without affecting HBV RNAs or cccDNA.

138 The specificity of our cccDNA probe set was confirmed by its strict intranuclea

139 ersion but inhibits the synthesis of progeny cccDNA by amplification.

140 alpha, inhibited HBV replication and reduced cccDNA in infected cells without the direct contact requ

141 at HBV envelope proteins negatively regulate cccDNA formation, and conversion of DP-rcDNA into cccDNA

142 e infected hepatocytes, are needed to remove cccDNA from surviving infected hepatocytes.

143 cytes by inducing deamination and subsequent cccDNA decay.

144 he development of drugs that directly target cccDNA is hampered by the lack of robust HBV cccDNA mode

145 Novel approaches that directly target cccDNA regulation would therefore be highly desirable.

146 at replicative DNA levels declined more than cccDNA and mRNA levels following adenovirus infection su

147 Collectively, these results suggest that cccDNA clearance is a two-step process mediated by the c

148 iral DNA found in the cytoplasm but also the cccDNA from the nucleus.

149 nstrate that the elimination kinetics of the cccDNA are more rapid than the elimination of HBV antige

150 In contrast, terminal clearance of the cccDNA is associated with the peak of liver disease and

151 ssion decreased rapidly-less than 50% of the cccDNA remained detectable in 1.5 days.

152 ylation as well as active recruitment to the cccDNA of transcriptional corepressors.

153 ificantly slowed the conversion of RC-DNA to cccDNA.

154 hat maintenance of wild-type levels of viral cccDNA promotes persistence of virus infection by establ

155 ed birds coincided with a reduction of viral cccDNA to wild-type virus levels in the liver.

156 ultiple enzymes which target different vital cccDNA regions, or sequential delivery of different enzy

157 he amount of hepatocyte turnover and whether cccDNA synthesis is effectively blocked during the perio

158 analysis of the recombination joints in WHV cccDNA.