Neonatal Lethality and Inflammatory Phenotype of the New Transgenic Mice with Overexpression of Human Interleukin-6 in Myeloid Cells

Зварцев, Р. В.; Korshunova, Diana S.; Горшкова, Е. А.; Носенко, М. А.; Korneev, Kirill V.; Maksimenko, Oksana; Коробко, И. В.; Kuprash, Dmitry V.; Drutskaya, Marina S.; Nedospasov, Sergei A.; Deikin, A. V.

doi:10.1134/s1607672918060157

Cited by 12 publications

(4 citation statements)

References 11 publications

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“…In the case of a new SARS-CoV-2 virus, it is impossible to predict its degree of contagion from mice to mice and human population. For the safety of the laboratory staff, we decided to use LoxP-induced expression of humanized genes, as described in (Zvartsev et al 2018;Deykin et al 2019;Silaeva et al 2020).…”

Section: Discussionmentioning

confidence: 99%

On the way from SARS-CoV-sensitive mice to murine COVID-19 model

Солдатов

Kubekina

Silaeva

et al. 2020

RRP

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The coronavirus disease 2019 (COVID-19) is a master killer which appeared suddenly and which has already claimed more than 200,000 human lives. In this situation, laboratories are in urgent need for a COVID-19 murine model to search for effective antiviral compounds. Here we propose a novel strategy for the development of mice that can be inoculated by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the COVID-19 causative agent. In humans, two proteins – ACE2 and TMPRSS2 – are involved in SARS-CoV-2 cells entry and, thus, we decided to introduce their genes into a murine genome. These genes will be placed with LoxP sites under the murine Tmprss2 promoter. Such an approach can provide a representative model with the opportunity to control the viral sensitivity of an animal population and tissue specificity of hACE2 and hTMPRSS2 expression. Graphical abstract The new COVID-19 model should be based on inducible co-expression of the human ACE2 and TMPRSS2 genes. Activation of ACE2 and TMPRSS2 genes will occur only in the virological laboratory, after crossbreeding with Cre-mice. Before activation, mice will be resistant to SARS-CoV-2 for their biological safety during the pandemic.

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Section: Discussionmentioning

confidence: 99%

On the way from SARS-CoV-sensitive mice to murine COVID-19 model

Солдатов

Kubekina

Silaeva

et al. 2020

RRP

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“…Loss of granulocyte–macrophage colony-stimulating factor results in highly expressed Il1b and Il6 in the spleen and increased mortality in mice [ 34 ]. Il6 also mediates recruiting leukocytes to the spleen and enhances Il6 level, causing early postnatal death [ 35 , 36 ]. Consistent with the results from previous reports, we observed that IL6 level was significantly increased in the spleen of Npr1 −/− mice.…”

Section: Discussionmentioning

confidence: 99%

Null Function of Npr1 Disturbs Immune Response in Colonic Inflammation During Early Postnatal Stage

et al. 2022

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Natriuretic peptide receptor 1 (NPR1) is conventionally known as a regulator of vascular homeostasis. Here, we generated an Npr1 knockout mouse model with CRISPR/Cas9 technology and found that homozygous mice (Npr1−/−) exhibited weight loss and poor survival rate during early postnatal stage. Careful examination revealed unexpectedly that Npr1−/− mice developed colitis characterized by shortened colon, evident colonic mucosal damage, increased histopathological score, and higher colonic expression of proinflammatory cytokines interleukin-1B (IL1B) and -6 (IL6). RNA-sequencing analysis revealed that differentially expressed genes were prominently enriched in the biological pathways related to immune response in both spleen and colon of Npr1−/− mice. Cytofluorimetric analysis demonstrated that leukocytes in the spleen were significantly increased, particularly, the populations of neutrophil and CD3+ T cell were elevated but CD4+ T cells were decreased in Npr1−/− mice. Administration of 8-Br-cGMP, a downstream activator of NPR1, restored these immune-cell populations disturbed in Npr1−/− mice and lessened the colitis-related phenotypes. To validate the involvement of Npr1 in colitis, we examined another mouse model induced by dextran sodium sulfate (DSS) and found a decreased Npr1 expression and shifted immune-cell populations as well. Importantly, 8-Br-cGMP treatment exhibited a similar effect in the restoration of immune-cell populations and attenuation of colonic inflammation in DSS mice. Our data indicate that loss of Npr1 possibly interrupts immune response, which is critical to the pathogenesis of colitis in the early life.

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“…Then the construction was prepared for microinjections: circular plasmid DNA was treated with the restriction enzyme PvuI (Thermo Fisher Scientific); the resulting fragments were separated by electrophoresis in agarose gel. A linear fragment 14832 bp, devoid of bacterial sequences, was extracted using a Monarch kit for isolation of DNA fragments from gels and reaction mixtures (New England Biolabs) and dissolved in TE buffer at a concentration of 1 ng/μl (Zvartsev et al 2019).…”

Section: Design and Manufacture Of Genetic Constructmentioning

confidence: 99%

“…Mitochondrial dysfunction (MD) represents an inadequate number of mitochondria, or an inability to provide necessary substrates to mitochondria, or a dysfunction in their electron transport and ATP-synthesis machinery (Nicolson 2014). Besides a large group of monogenic mitochondrial diseases (MD) is a key link in the pathobiology of a wide spectrum of disorders, including cardiovascular (Zakirov et al 2020), neurodegenerative (Angelova et al 2021) and metabolic diseases (Prasun 2020).…”

Section: Introductionmentioning

confidence: 99%

Transgenic mice Cre-dependently expressing mutant polymerase-gamma: novel test-system for pharmacological study of mitoprotective drugs

Kubekina¹,

Silaeva²,

Bruter³

et al. 2021

RRP

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Introduction: PolG-alpha is a nuclear-encoded enzyme which provides replication and repair of mitochondrial DNA. D257A mutation of PolG-alpha leads to change in the N-terminal ”proofreading” domain, which deprives the enzyme of 3′-5′ exonuclease activity, resulting in accumulation of mutations in the mitochondrial genome. Materials and methods: Murine zygotes were microinjected with transgene construction carrying mutant murine Polg coding sequence and GFP coding sequence by a loxP-flanked STOP-cassette. Two Cre-activator strains, CMV-Cre (systemic activation) and Tie2-Cre (endothelial activation), were used for activation of the transgene. To confirm the insertion and Cre-dependent activation of the transgene, genotyping and qPCR copy number measurement of mutant Polg were performed, and GFP fluorescence was assessed. Results: Two primary transgenic animals were used as the founders for two lines with copy numbers of transgene ~7 and ~5. After systemic activation, the number of the transgene copies decreases to ~1.0 while endothelial specific activation does not affect the number of transgene copies in tail tissue. Discussion: A murine model with spatial control of mutant Polg expression has been developed. To our knowledge, this is the first transgenic model of tissue-specific mitochondrial dysfunction. Conclusion: Transgenic mice Cre-dependent expressing mutant polymerase-gamma are a novel test-system for studying mitochondrial biology and efficacy of mitoprotective drugs.

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Neonatal Lethality and Inflammatory Phenotype of the New Transgenic Mice with Overexpression of Human Interleukin-6 in Myeloid Cells

Cited by 12 publications

References 11 publications

On the way from SARS-CoV-sensitive mice to murine COVID-19 model

On the way from SARS-CoV-sensitive mice to murine COVID-19 model

Null Function of Npr1 Disturbs Immune Response in Colonic Inflammation During Early Postnatal Stage

Transgenic mice Cre-dependently expressing mutant polymerase-gamma: novel test-system for pharmacological study of mitoprotective drugs

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