A Herpes Simplex Virus 1 DNA Polymerase Multidrug Resistance Mutation Identified in a Patient With Immunodeficiency and Confirmed by Gene Editing
© The Author(s) 2023. Published by Oxford University Press on behalf of Infectious Diseases Society of America. All rights reserved. For permissions, please e-mail: journals.permissionsoup.com..
BACKGROUND: Herpes simplex virus 1 can cause severe infections in individuals who are immunocompromised. In these patients, emergence of drug resistance mutations causes difficulties in infection management.
METHODS: Seventeen herpes simplex virus 1 isolates were obtained from orofacial/anogenital lesions in a patient with leaky severe combined immunodeficiency over 7 years, before and after stem cell transplantation. Spatial/temporal evolution of drug resistance was characterized genotypically-with Sanger and next-generation sequencing of viral thymidine kinase (TK) and DNA polymerase (DP)-and phenotypically. CRISPR/Cas9 was used to introduce the novel DP Q727R mutation, and dual infection-competition assays were performed to assess viral fitness.
RESULTS: Isolates had identical genetic backgrounds, suggesting that orofacial/anogenital infections derived from the same virus lineage. Eleven isolates proved heterogeneous TK virus populations by next-generation sequencing, undetectable by Sanger sequencing. Thirteen isolates were acyclovir resistant due to TK mutations, and the Q727R isolate additionally exhibited foscarnet/adefovir resistance. Recombinant Q727R mutant virus showed multidrug resistance and increased fitness under antiviral pressure.
CONCLUSIONS: Long-term follow-up of a patient with severe combined immunodeficiency revealed virus evolution and frequent reactivation of wild-type and TK mutant strains, mostly as heterogeneous populations. The DP Q727R resistance phenotype was confirmed with CRISPR/Cas9, a useful tool to validate novel drug resistance mutations.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:228 |
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| Enthalten in: |
The Journal of infectious diseases - 228(2023), 11 vom: 28. Nov., Seite 1505-1515 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Schalkwijk, Hanna Helena [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 29.11.2023 Date Revised 06.02.2024 published: Print Citation Status MEDLINE |
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| doi: |
10.1093/infdis/jiad184 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM357269101 |
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| 100 | 1 | |a Schalkwijk, Hanna Helena |e verfasserin |4 aut | |
| 245 | 1 | 2 | |a A Herpes Simplex Virus 1 DNA Polymerase Multidrug Resistance Mutation Identified in a Patient With Immunodeficiency and Confirmed by Gene Editing |
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| 500 | |a Date Revised 06.02.2024 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a © The Author(s) 2023. Published by Oxford University Press on behalf of Infectious Diseases Society of America. All rights reserved. For permissions, please e-mail: journals.permissionsoup.com. | ||
| 520 | |a BACKGROUND: Herpes simplex virus 1 can cause severe infections in individuals who are immunocompromised. In these patients, emergence of drug resistance mutations causes difficulties in infection management | ||
| 520 | |a METHODS: Seventeen herpes simplex virus 1 isolates were obtained from orofacial/anogenital lesions in a patient with leaky severe combined immunodeficiency over 7 years, before and after stem cell transplantation. Spatial/temporal evolution of drug resistance was characterized genotypically-with Sanger and next-generation sequencing of viral thymidine kinase (TK) and DNA polymerase (DP)-and phenotypically. CRISPR/Cas9 was used to introduce the novel DP Q727R mutation, and dual infection-competition assays were performed to assess viral fitness | ||
| 520 | |a RESULTS: Isolates had identical genetic backgrounds, suggesting that orofacial/anogenital infections derived from the same virus lineage. Eleven isolates proved heterogeneous TK virus populations by next-generation sequencing, undetectable by Sanger sequencing. Thirteen isolates were acyclovir resistant due to TK mutations, and the Q727R isolate additionally exhibited foscarnet/adefovir resistance. Recombinant Q727R mutant virus showed multidrug resistance and increased fitness under antiviral pressure | ||
| 520 | |a CONCLUSIONS: Long-term follow-up of a patient with severe combined immunodeficiency revealed virus evolution and frequent reactivation of wild-type and TK mutant strains, mostly as heterogeneous populations. The DP Q727R resistance phenotype was confirmed with CRISPR/Cas9, a useful tool to validate novel drug resistance mutations | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a CRISPR/Cas9 precise gene editing | |
| 650 | 4 | |a drug resistance | |
| 650 | 4 | |a hematopoietic stem cell transplantation | |
| 650 | 4 | |a herpes simplex virus 1 | |
| 650 | 4 | |a viral evolution | |
| 650 | 7 | |a Antiviral Agents |2 NLM | |
| 650 | 7 | |a Acyclovir |2 NLM | |
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| 650 | 7 | |a DNA-Directed DNA Polymerase |2 NLM | |
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| 650 | 7 | |a Thymidine Kinase |2 NLM | |
| 650 | 7 | |a EC 2.7.1.21 |2 NLM | |
| 700 | 1 | |a Georgala, Aspasia |e verfasserin |4 aut | |
| 700 | 1 | |a Gillemot, Sarah |e verfasserin |4 aut | |
| 700 | 1 | |a Temblador, Arturo |e verfasserin |4 aut | |
| 700 | 1 | |a Topalis, Dimitri |e verfasserin |4 aut | |
| 700 | 1 | |a Wittnebel, Sebastian |e verfasserin |4 aut | |
| 700 | 1 | |a Andrei, Graciela |e verfasserin |4 aut | |
| 700 | 1 | |a Snoeck, Robert |e verfasserin |4 aut | |
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