In vivo repair of rat liver DNA damaged by dimethylnitrosamine or diethylnitrosamine

Engelse, L. Den; Philippus, E.J.

doi:10.1016/0009-2797(77)90046-1

Cited by 22 publications

(5 citation statements)

References 22 publications

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“…On the basis of the promising results with this first experiment, a second experiment was carried out with the following modifications: Instead of NNM, the ethylating agent DENA was used because this carcinogen is known to give rise to a relatively large proportion of 0 6 -guanine ethylations (8] kn1J1wn for their miscoding properties [9]. The doses used were much lower so as not to induce any liver darnage [10].…”

Section: Resultsmentioning

confidence: 99%

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In vivo assay for somatic point mutations induced by genotoxic carcinogens: Incorporation of [35S]methionine into a rat liver cytochrome b5 normally lacking sulphur-containing amino acids

Jauch

Lutz

1983

Chemico-Biological Interactions

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Section: Resultsmentioning

confidence: 99%

“…Of the total number of ethylations, only about 9o/C are at ~he 0 6 of guanine. resulting~ in a critical premutagenic lesion [8]. Taking have grown into a group of mutated progeny or even into a hyperplastic nodule ol · .…”

Section: Resultsmentioning

confidence: 99%

In vivo assay for somatic point mutations induced by genotoxic carcinogens: Incorporation of [35S]methionine into a rat liver cytochrome b5 normally lacking sulphur-containing amino acids

Jauch

Lutz

1983

Chemico-Biological Interactions

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“…Mice treated with diethylnitrosamine (DEN), a potent mutagen, showed intense staining (Extended Fig. 12c-d) 34 . During udarabine treatment, mouse livers possessed widespread pan-nuclear γH2AX foci (Fig.…”

Section: Fludarabine Induced a Transient Dna Damage Responsementioning

confidence: 99%

Fludarabine increases nuclease-free AAV- and CRISPR/Cas9-mediated homologous recombination in mice

et al. 2022

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Recombinant adeno-associated viral (rAAV) vectors continue to gain popularity for in vivo therapeutic gene delivery. Homologous recombination-based gene therapy using rAAV (AAV-HR) without nucleases has several advantages over classical gene therapy, especially when targeting the liver in neonatal/pediatric populations due to its potential for permanent sustained transgene expression.However, the low e ciency of AAV-HR remains a limitation for some clinical applications. Here, we tested series of small molecule compounds with different mechanisms of action in the context of AAV-HR and identi ed that ribonucleotide reductase (RNR) inhibitors signi cantly enhanced the AAV-HR e ciency in mouse and human liver cell lines. Furthermore, short term administration of the RNR inhibitor udarabine increased the in vivo e ciency of both non-nuclease and CRISPR/Cas9-mediated AAV-HR in the murine liver, without causing toxicity. Mechanistic experiments showed that udarabine administration induced transient DNA damage signaling in both proliferating and quiescent hepatocytes. Surprisingly, in vivo BrdU labeling implicated that the majority of AAV-HR events occurred in non-proliferating hepatocytes in both udarabine-treated and control mice. These studies suggested that the induction of transient DNA repair signaling in non-dividing hepatocytes was responsible for enhancing the e ciency of AAV-HR in mice during RNR inhibition. In total, we show the use of a clinically approved RNR inhibitor can enhance AAV-HR based genome editing therapeutics. Contributions S.T. and M.A.K. designed the study. S.T., C.J.S and M.A.K. reviewed all data and wrote paper with all coauthors. S.T. performed in vitro studies. S.T., C.J.S. and F.Z. performed in vivo studies. C.J.S. performed imaging analysis. G.B. and A.F.M. performed in vivo study using CRISPR/Cas9 system. S.T., K.P. and G.A. designed and made plasmids. S.T. and K.P. performed rAAV production. H.J. performed ALT assay.

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“…Minimal levels of γH2AX were detected in untreated or BrdU-only treated livers ( Fig.4a-b). Mice treated with diethylnitrosamine (DEN), a potent mutagen, showed intense staining (Extended Fig.12c-d) 34 . During udarabine treatment, mouse livers possessed widespread pan-nuclear γH2AX foci ( Fig.4a-b).…”

Section: Fludarabine Administration Increased Aav-mediated Gene Targementioning

confidence: 99%

Improving the efficiency of liver targeting rAAV-mediated homologous recombination using ribonucleotide reductase inhibitors

Tsuji¹,

C²,

Bortolussi³

et al. 2020

Preprint

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Recombinant adeno-associated viral (rAAV) vectors continue to gain popularity for in vivo therapeutic gene delivery. Homologous recombination-based gene therapy using rAAV (AAV-HR) without nucleases has several advantages over classical gene therapy, especially when targeting the liver in neonatal/pediatric populations due to its potential for permanent sustained transgene expression. However, the low efficiency of AAV-HR remains a limitation for some clinical applications. Here, we tested series of small molecule compounds with different mechanisms of action in the context of AAV-HR and identified that ribonucleotide reductase (RNR) inhibitors significantly enhanced the AAV-HR efficiency in mouse and human liver cell lines. Furthermore, short term administration of the RNR inhibitor fludarabine increased the in vivo efficiency of both non-nuclease and CRISPR/Cas9-mediated AAV-HR in the murine liver, without causing toxicity. Mechanistic experiments showed that fludarabine administration induced transient DNA damage signaling in both proliferating and quiescent hepatocytes. Surprisingly, in vivo BrdU labeling implicated that the majority of AAV-HR events occurred in non-proliferating hepatocytes in both fludarabine-treated and control mice. These studies suggested that the induction of transient DNA repair signaling in non-dividing hepatocytes was responsible for enhancing the efficiency of AAV-HR in mice during RNR inhibition. In total, we show the use of a clinically approved RNR inhibitor can enhance AAV-HR based genome editing therapeutics.

show abstract

In vivo repair of rat liver DNA damaged by dimethylnitrosamine or diethylnitrosamine

Cited by 22 publications

References 22 publications

In vivo assay for somatic point mutations induced by genotoxic carcinogens: Incorporation of [35S]methionine into a rat liver cytochrome b5 normally lacking sulphur-containing amino acids

In vivo assay for somatic point mutations induced by genotoxic carcinogens: Incorporation of [35S]methionine into a rat liver cytochrome b5 normally lacking sulphur-containing amino acids

Fludarabine increases nuclease-free AAV- and CRISPR/Cas9-mediated homologous recombination in mice

Improving the efficiency of liver targeting rAAV-mediated homologous recombination using ribonucleotide reductase inhibitors

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