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  1. Article ; Online: Overlapping group screening for detection of gene-environment interactions with application to TCGA high-dimensional survival genomic data.

    Wang, Jie-Huei / Wang, Kang-Hsin / Chen, Yi-Hau

    BMC bioinformatics

    2022  Volume 23, Issue 1, Page(s) 202

    Abstract: Background: In the context of biomedical and epidemiological research, gene-environment (G-E) interaction is of great significance to the etiology and progression of many complex diseases. In high-dimensional genetic data, two general models, marginal ... ...

    Abstract Background: In the context of biomedical and epidemiological research, gene-environment (G-E) interaction is of great significance to the etiology and progression of many complex diseases. In high-dimensional genetic data, two general models, marginal and joint models, are proposed to identify important interaction factors. Most existing approaches for identifying G-E interactions are limited owing to the lack of robustness to outliers/contamination in response and predictor data. In particular, right-censored survival outcomes make the associated feature screening even challenging. In this article, we utilize the overlapping group screening (OGS) approach to select important G-E interactions related to clinical survival outcomes by incorporating the gene pathway information under a joint modeling framework.
    Results: Simulation studies under various scenarios are carried out to compare the performances of our proposed method with some commonly used methods. In the real data applications, we use our proposed method to identify G-E interactions related to the clinical survival outcomes of patients with head and neck squamous cell carcinoma, and esophageal carcinoma in The Cancer Genome Atlas clinical survival genetic data, and further establish corresponding survival prediction models. Both simulation and real data studies show that our method performs well and outperforms existing methods in the G-E interaction selection, effect estimation, and survival prediction accuracy.
    Conclusions: The OGS approach is useful for selecting important environmental factors, genes and G-E interactions in the ultra-high dimensional feature space. The prediction ability of OGS with the Lasso penalty is better than existing methods. The same idea of the OGS approach can apply to other outcome models, such as the proportional odds survival time model, the logistic regression model for binary outcomes, and the multinomial logistic regression model for multi-class outcomes.
    MeSH term(s) Computer Simulation ; Gene-Environment Interaction ; Genomics ; Humans ; Neoplasms/genetics ; Research
    Language English
    Publishing date 2022-05-30
    Publishing country England
    Document type Journal Article
    ZDB-ID 2041484-5
    ISSN 1471-2105 ; 1471-2105
    ISSN (online) 1471-2105
    ISSN 1471-2105
    DOI 10.1186/s12859-022-04750-7
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article: KSHV vIL-6 Enhances Inflammatory Responses by Epigenetic Reprogramming.

    Inagaki, Tomoki / Wang, Kang-Hsin / Kumar, Ashish / Izumiya, Chie / Miura, Hiroki / Komaki, Somayeh / Davis, Ryan R / Tepper, Clifford G / Katano, Harutaka / Shimoda, Michiko / Izumiya, Yoshihiro

    bioRxiv : the preprint server for biology

    2023  

    Abstract: Kaposi sarcoma-associated herpesvirus (KSHV) inflammatory cytokine syndrome (KICS) is a newly described chronic inflammatory disease condition caused by KSHV infection and is characterized by high KSHV viral load and sustained elevations of serum KSHV- ... ...

    Abstract Kaposi sarcoma-associated herpesvirus (KSHV) inflammatory cytokine syndrome (KICS) is a newly described chronic inflammatory disease condition caused by KSHV infection and is characterized by high KSHV viral load and sustained elevations of serum KSHV-encoded IL-6 (vIL-6) and human IL-6 (hIL-6). KICS has significant immortality and possesses greater risks of having other complications, which include malignancies. Although prolonged inflammatory vIL-6 exposure by persistent KSHV infection is expected to have key roles in subsequent disease development, the biological effects of prolonged vIL-6 exposure remain elusive. Using thiol-Linked Alkylation for the Metabolic Sequencing and Cleavage Under Target & Release Using Nuclease analysis, we studied the effect of prolonged vIL-6 exposure in chromatin landscape and resulting cytokine production. The studies showed that prolonged vIL-6 exposure increased Bromodomain containing 4 (BRD4) and histone H3 lysine 27 acetylation co-occupancies on chromatin, and the recruitment sites were frequently co-localized with poised RNAPII with associated enzymes. Increased BRD4 recruitment on promoters was associated with increased and prolonged NF-κB p65 binding after the lipopolysaccharide stimulation. The p65 binding resulted in quicker and sustained transcription bursts from the promoters; this mechanism increased total amounts of hIL-6 and IL-10 in tissue culture. Pretreatment with the BRD4 inhibitor, OTX015, eliminated the enhanced inflammatory cytokine production. These findings suggest that persistent vIL-6 exposure may establish a chromatin landscape favorable for the reactivation of inflammatory responses in monocytes. This epigenetic memory may explain the greater risk of chronic inflammatory disease development in KSHV-infected individuals.
    Language English
    Publishing date 2023-07-23
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2023.06.25.546454
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: KSHV vIL-6 enhances inflammatory responses by epigenetic reprogramming.

    Inagaki, Tomoki / Wang, Kang-Hsin / Kumar, Ashish / Izumiya, Chie / Miura, Hiroki / Komaki, Somayeh / Davis, Ryan R / Tepper, Clifford G / Katano, Harutaka / Shimoda, Michiko / Izumiya, Yoshihiro

    PLoS pathogens

    2023  Volume 19, Issue 11, Page(s) e1011771

    Abstract: Kaposi sarcoma-associated herpesvirus (KSHV) inflammatory cytokine syndrome (KICS) is a newly described chronic inflammatory disease condition caused by KSHV infection and is characterized by high KSHV viral load and sustained elevations of serum KSHV- ... ...

    Abstract Kaposi sarcoma-associated herpesvirus (KSHV) inflammatory cytokine syndrome (KICS) is a newly described chronic inflammatory disease condition caused by KSHV infection and is characterized by high KSHV viral load and sustained elevations of serum KSHV-encoded IL-6 (vIL-6) and human IL-6 (hIL-6). KICS has significant immortality and greater risks of other complications, including malignancies. Although prolonged inflammatory vIL-6 exposure by persistent KSHV infection is expected to have key roles in subsequent disease development, the biological effects of prolonged vIL-6 exposure remain elusive. Using thiol(SH)-linked alkylation for the metabolic (SLAM) sequencing and Cleavage Under Target & Release Using Nuclease analysis (CUT&RUN), we studied the effect of prolonged vIL-6 exposure in chromatin landscape and resulting cytokine production. The studies showed that prolonged vIL-6 exposure increased Bromodomain containing 4 (BRD4) and histone H3 lysine 27 acetylation co-occupancies on chromatin, and the recruitment sites were frequently co-localized with poised RNA polymerase II with associated enzymes. Increased BRD4 recruitment on promoters was associated with increased and prolonged NF-κB p65 binding after the lipopolysaccharide stimulation. The p65 binding resulted in quicker and sustained transcription bursts from the promoters; this mechanism increased total amounts of hIL-6 and IL-10 in tissue culture. Pretreatment with the BRD4 inhibitors, OTX015 and MZ1, eliminated the enhanced inflammatory cytokine production. These findings suggest that persistent vIL-6 exposure may establish a chromatin landscape favorable for the reactivation of inflammatory responses in monocytes. This epigenetic memory may explain the greater risk of chronic inflammatory disease development in KSHV-infected individuals.
    MeSH term(s) Humans ; Herpesvirus 8, Human/physiology ; Interleukin-6/metabolism ; Nuclear Proteins/metabolism ; Transcription Factors/metabolism ; Cytokines/metabolism ; Herpesviridae Infections/metabolism ; Chromatin/metabolism ; Epigenesis, Genetic ; Sarcoma, Kaposi ; Cell Cycle Proteins/metabolism
    Chemical Substances Interleukin-6 ; Nuclear Proteins ; Transcription Factors ; Cytokines ; Chromatin ; BRD4 protein, human ; Cell Cycle Proteins
    Language English
    Publishing date 2023-11-07
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2205412-1
    ISSN 1553-7374 ; 1553-7374
    ISSN (online) 1553-7374
    ISSN 1553-7374
    DOI 10.1371/journal.ppat.1011771
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Determination of the Maturation Status of Dendritic Cells by Applying Pattern Recognition to High-Resolution Images.

    Lohrer, Michael F / Liu, Yang / Hanna, Darrin M / Wang, Kang-Hsin / Liu, Fu-Tong / Laurence, Ted A / Liu, Gang-Yu

    The journal of physical chemistry. B

    2020  Volume 124, Issue 39, Page(s) 8540–8548

    Abstract: The maturation or activation status of dendritic cells (DCs) directly correlates with their behavior and immunofunction. A common means to determine the maturity of dendritic cells is from high-resolution images acquired via scanning electron microscopy ( ...

    Abstract The maturation or activation status of dendritic cells (DCs) directly correlates with their behavior and immunofunction. A common means to determine the maturity of dendritic cells is from high-resolution images acquired via scanning electron microscopy (SEM) or atomic force microscopy (AFM). While direct and visual, the determination has been made by directly looking at the images by researchers. This work reports a machine learning approach using pattern recognition in conjunction with cellular biophysical knowledge of dendritic cells to determine the maturation status of dendritic cells automatically. The determination from AFM images reaches 100% accuracy. The results from SEM images reaches 94.9%. The results demonstrate the accuracy of using machine learning for accelerating data analysis, extracting information, and drawing conclusions from high-resolution cellular images, paving the way for future applications requiring high-throughput and automation, such as cellular sorting and selection based on morphology, quantification of cellular structure, and DC-based immunotherapy.
    MeSH term(s) Dendritic Cells ; Machine Learning ; Microscopy, Atomic Force ; Microscopy, Electron, Scanning
    Language English
    Publishing date 2020-09-22
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
    ISSN 1520-5207
    ISSN (online) 1520-5207
    DOI 10.1021/acs.jpcb.0c06437
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article: Determination of the Maturation Status of Dendritic Cells by Applying Pattern Recognition to High-Resolution Images

    Lohrer, Michael F / Liu, Yang / Hanna, Darrin M / Wang, Kang-Hsin / Liu, Fu-Tong / Laurence, Ted A / Liu, Gang-yu

    Journal of physical chemistry. 2020 Sept. 03, v. 124, no. 39

    2020  

    Abstract: The maturation or activation status of dendritic cells (DCs) directly correlates with their behavior and immunofunction. A common means to determine the maturity of dendritic cells is from high-resolution images acquired via scanning electron microscopy ( ...

    Abstract The maturation or activation status of dendritic cells (DCs) directly correlates with their behavior and immunofunction. A common means to determine the maturity of dendritic cells is from high-resolution images acquired via scanning electron microscopy (SEM) or atomic force microscopy (AFM). While direct and visual, the determination has been made by directly looking at the images by researchers. This work reports a machine learning approach using pattern recognition in conjunction with cellular biophysical knowledge of dendritic cells to determine the maturation status of dendritic cells automatically. The determination from AFM images reaches 100% accuracy. The results from SEM images reaches 94.9%. The results demonstrate the accuracy of using machine learning for accelerating data analysis, extracting information, and drawing conclusions from high-resolution cellular images, paving the way for future applications requiring high-throughput and automation, such as cellular sorting and selection based on morphology, quantification of cellular structure, and DC-based immunotherapy.
    Keywords atomic force microscopy ; automation ; cell structures ; immunotherapy ; physical chemistry
    Language English
    Dates of publication 2020-0903
    Size p. 8540-8548.
    Publishing place American Chemical Society
    Document type Article
    Note NAL-AP-2-clean
    ISSN 1520-5207
    DOI 10.1021/acs.jpcb.0c06437
    Database NAL-Catalogue (AGRICOLA)

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  6. Article ; Online: KSHV episome tethering sites on host chromosomes and regulation of latency-lytic switch by CHD4.

    Kumar, Ashish / Lyu, Yuanzhi / Yanagihashi, Yuichi / Chantarasrivong, Chanikarn / Majerciak, Vladimir / Salemi, Michelle / Wang, Kang-Hsin / Inagaki, Tomoki / Chuang, Frank / Davis, Ryan R / Tepper, Clifford G / Nakano, Kazushi / Izumiya, Chie / Shimoda, Michiko / Nakajima, Ken-Ichi / Merleev, Alexander / Zheng, Zhi-Ming / Campbell, Mel / Izumiya, Yoshihiro

    Cell reports

    2022  Volume 39, Issue 6, Page(s) 110788

    Abstract: Kaposi sarcoma-associated herpesvirus (KSHV) establishes a latent infection in the cell nucleus, but where KSHV episomal genomes are tethered and the mechanisms underlying KSHV lytic reactivation are unclear. Here, we study the nuclear microenvironment ... ...

    Abstract Kaposi sarcoma-associated herpesvirus (KSHV) establishes a latent infection in the cell nucleus, but where KSHV episomal genomes are tethered and the mechanisms underlying KSHV lytic reactivation are unclear. Here, we study the nuclear microenvironment of KSHV episomes and show that the KSHV latency-lytic replication switch is regulated via viral long non-coding (lnc)RNA-CHD4 (chromodomain helicase DNA binding protein 4) interaction. KSHV episomes localize with CHD4 and ADNP proteins, components of the cellular ChAHP complex. The CHD4 and ADNP proteins occupy the 5'-region of the highly inducible lncRNAs and terminal repeats of the KSHV genome together with latency-associated nuclear antigen (LANA). Viral lncRNA binding competes with CHD4 DNA binding, and KSHV reactivation sequesters CHD4 from the KSHV genome, which is also accompanied by detachment of KSHV episomes from host chromosome docking sites. We propose a model in which robust KSHV lncRNA expression determines the latency-lytic decision by regulating LANA/CHD4 binding to KSHV episomes.
    MeSH term(s) Antigens, Viral/genetics ; Antigens, Viral/metabolism ; Chromosomes/metabolism ; Herpesvirus 8, Human/genetics ; Humans ; Mi-2 Nucleosome Remodeling and Deacetylase Complex/genetics ; Plasmids ; RNA, Long Noncoding/genetics ; Sarcoma, Kaposi ; Tumor Microenvironment ; Virus Latency/genetics
    Chemical Substances Antigens, Viral ; CHD4 protein, human ; RNA, Long Noncoding ; Mi-2 Nucleosome Remodeling and Deacetylase Complex (EC 3.5.1.98)
    Language English
    Publishing date 2022-05-06
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, N.I.H., Intramural
    ZDB-ID 2649101-1
    ISSN 2211-1247 ; 2211-1247
    ISSN (online) 2211-1247
    ISSN 2211-1247
    DOI 10.1016/j.celrep.2022.110788
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: KSHV transactivator-derived small peptide traps coactivators to attenuate MYC and inhibits leukemia and lymphoma cell growth.

    Shimoda, Michiko / Lyu, Yuanzhi / Wang, Kang-Hsin / Kumar, Ashish / Miura, Hiroki / Meckler, Joshua F / Davis, Ryan R / Chantarasrivong, Chanikarn / Izumiya, Chie / Tepper, Clifford G / Nakajima, Ken-Ichi / Tuscano, Joseph / Barisone, Gustavo / Izumiya, Yoshihiro

    Communications biology

    2021  Volume 4, Issue 1, Page(s) 1330

    Abstract: In herpesvirus replicating cells, host cell gene transcription is frequently down-regulated because important transcriptional apparatuses are appropriated by viral transcription factors. Here, we show a small peptide derived from the Kaposi's sarcoma- ... ...

    Abstract In herpesvirus replicating cells, host cell gene transcription is frequently down-regulated because important transcriptional apparatuses are appropriated by viral transcription factors. Here, we show a small peptide derived from the Kaposi's sarcoma-associated herpesvirus transactivator (K-Rta) sequence, which attenuates cellular MYC expression, reduces cell proliferation, and selectively kills cancer cell lines in both tissue culture and a xenograft tumor mouse model. Mechanistically, the peptide functions as a decoy to block the recruitment of coactivator complexes consisting of Nuclear receptor coactivator 2 (NCOA2), p300, and SWI/SNF proteins to the MYC promoter in primary effusion lymphoma cells. Thiol(SH)-linked alkylation for the metabolic sequencing of RNA (SLAM seq) with target-transcriptional analyses further confirm that the viral peptide directly attenuates MYC and MYC-target gene expression. This study thus provides a unique tool to control MYC activation, which may be used as a therapeutic payload to treat MYC-dependent diseases such as cancers and autoimmune diseases.
    MeSH term(s) Cell Line, Tumor ; Cell Proliferation ; Herpesvirus 8, Human/chemistry ; Herpesvirus 8, Human/physiology ; Humans ; Leukemia/physiopathology ; Lymphoma/physiopathology ; Proto-Oncogene Proteins c-myc/genetics ; Proto-Oncogene Proteins c-myc/metabolism ; Trans-Activators/genetics ; Trans-Activators/metabolism ; Tumor Cells, Cultured
    Chemical Substances Proto-Oncogene Proteins c-myc ; Trans-Activators
    Language English
    Publishing date 2021-12-02
    Publishing country England
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ISSN 2399-3642
    ISSN (online) 2399-3642
    DOI 10.1038/s42003-021-02853-0
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  8. Article ; Online: Proximity Biotin Labeling Reveals Kaposi's Sarcoma-Associated Herpesvirus Interferon Regulatory Factor Networks.

    Kumar, Ashish / Salemi, Michelle / Bhullar, Resham / Guevara-Plunkett, Sara / Lyu, Yuanzhi / Wang, Kang-Hsin / Izumiya, Chie / Campbell, Mel / Nakajima, Ken-Ichi / Izumiya, Yoshihiro

    Journal of virology

    2021  Volume 95, Issue 9

    Abstract: Studies on "hit-and-run" effects by viral proteins are difficult when using traditional affinity precipitation-based techniques under dynamic conditions, because only proteins interacting at a specific instance in time can be precipitated by affinity ... ...

    Abstract Studies on "hit-and-run" effects by viral proteins are difficult when using traditional affinity precipitation-based techniques under dynamic conditions, because only proteins interacting at a specific instance in time can be precipitated by affinity purification. Recent advances in proximity labeling (PL) have enabled identification of both static and dynamic protein-protein interactions. In this study, we applied a PL method by generating recombinant Kaposi's sarcoma-associated herpesvirus (KSHV). KSHV, a gammaherpesvirus, uniquely encodes four interferon regulatory factors (IRF-1 to -4) that suppress host interferon responses, and we examined KSHV IRF-1 and IRF-4 neighbor proteins to identify cellular proteins involved in innate immune regulation. PL identified 213 and 70 proteins as neighboring proteins of viral IRF-1 (vIRF-1) and vIRF-4 during viral reactivation, and 47 proteins were shared between the two vIRFs; the list also includes three viral proteins, ORF17, thymidine kinase, and vIRF-4. Functional annotation of respective interacting proteins showed highly overlapping biological roles such as mRNA processing and transcriptional regulation by TP53. Innate immune regulation by these commonly interacting 44 cellular proteins was examined with small interfering RNAs (siRNAs), and the splicing factor 3B family proteins were found to be associated with interferon transcription and to act as suppressors of KSHV reactivation. We propose that recombinant mini-TurboID-KSHV is a powerful tool to probe key cellular proteins that play a role in KSHV replication and that selective splicing factors have a function in the regulation of innate immune responses.
    MeSH term(s) Biotinylation/methods ; Gene Expression Regulation, Viral ; HEK293 Cells ; Herpesvirus 8, Human/metabolism ; Host Microbial Interactions ; Humans ; Interferon Regulatory Factors/metabolism ; Proteomics ; Sarcoma, Kaposi/virology ; Viral Proteins/metabolism ; Virus Replication
    Chemical Substances Interferon Regulatory Factors ; Viral Proteins ; viral interferon regulatory factors
    Language English
    Publishing date 2021-04-12
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 80174-4
    ISSN 1098-5514 ; 0022-538X
    ISSN (online) 1098-5514
    ISSN 0022-538X
    DOI 10.1128/JVI.02049-20
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  9. Article ; Online: IL-10 is overexpressed in human cutaneous T-cell lymphoma and is required for maximal tumor growth in a mouse model.

    Wu, Xuesong / Hsu, Daniel K / Wang, Kang-Hsin / Huang, Yuanshen / Mendoza, Lindsay / Zhou, Youwen / Hwang, Sam T

    Leukemia & lymphoma

    2018  Volume 60, Issue 5, Page(s) 1244–1252

    Abstract: A crucial question pertains to a role of IL-10 as a tumorigenic factor, or just a marker of advanced disease in cutaneous T-cell lymphoma (CTCL). Herein, we measured significantly elevated IL-10 mRNA in a cohort of skin samples of patients with CTCL. ... ...

    Abstract A crucial question pertains to a role of IL-10 as a tumorigenic factor, or just a marker of advanced disease in cutaneous T-cell lymphoma (CTCL). Herein, we measured significantly elevated IL-10 mRNA in a cohort of skin samples of patients with CTCL. Increased IL-10 was also detected in the tumor microenvironment of an established inflammation-dependent murine model of using MBL2 T lymphoma cells. Conditioned media from MBL2 cells was able to stimulate IL-10 production in bone marrow-derived macrophages in an IL-4-dependent manner. Implanted MBL2 T-cell lymphomas in IL-10KO mice were 50% smaller, accompanied by decreased numbers of infiltrating macrophages and reduced efficiency of M2-polarization compared with wild-type mice. With anti-IL-10R mAb treatment, both wild-type tumor-bearing mice and IL-10KO mice exhibited a further growth inhibition. Our data indicate that targeting IL-10 signaling with neutralizing antibodies to IL-10 or its receptor may have a great potential for advanced CTCL therapy.
    MeSH term(s) Animals ; Biopsy ; Cell Line, Tumor ; Cell Proliferation ; Disease Models, Animal ; Female ; Gene Expression ; Humans ; Interleukin-10/genetics ; Interleukin-10/metabolism ; Lymphoma, T-Cell, Cutaneous/genetics ; Lymphoma, T-Cell, Cutaneous/metabolism ; Lymphoma, T-Cell, Cutaneous/pathology ; Macrophages/immunology ; Macrophages/metabolism ; Macrophages/pathology ; Mice ; Neoplasm Staging ; Signal Transduction ; Tumor Burden ; Tumor Microenvironment/genetics ; Tumor Microenvironment/immunology ; Xenograft Model Antitumor Assays
    Chemical Substances IL10 protein, human ; Interleukin-10 (130068-27-8)
    Language English
    Publishing date 2018-10-02
    Publishing country United States
    Document type Journal Article
    ZDB-ID 1042374-6
    ISSN 1029-2403 ; 1042-8194
    ISSN (online) 1029-2403
    ISSN 1042-8194
    DOI 10.1080/10428194.2018.1516037
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article ; Online: Rainbow Kaposi's Sarcoma-Associated Herpesvirus Revealed Heterogenic Replication with Dynamic Gene Expression.

    Nakajima, Ken-Ichi / Guevara-Plunkett, Sara / Chuang, Frank / Wang, Kang-Hsin / Lyu, Yuanzhi / Kumar, Ashish / Luxardi, Guillaume / Izumiya, Chie / Soulika, Athena / Campbell, Mel / Izumiya, Yoshihiro

    Journal of virology

    2020  Volume 94, Issue 8

    Abstract: Molecular mechanisms of Kaposi's sarcoma-associated herpesvirus (KSHV) reactivation have been studied primarily by measuring the total or average activity of an infected cell population, which often consists of a mixture of both nonresponding and ... ...

    Abstract Molecular mechanisms of Kaposi's sarcoma-associated herpesvirus (KSHV) reactivation have been studied primarily by measuring the total or average activity of an infected cell population, which often consists of a mixture of both nonresponding and reactivating cells that in turn contain KSHVs at various stages of replication. Studies on KSHV gene regulation at the individual cell level would allow us to better understand the basis for this heterogeneity, and new preventive measures could be developed based on findings from nonresponding cells exposed to reactivation stimuli. Here, we generated a recombinant reporter virus, which we named "Rainbow-KSHV," that encodes three fluorescence-tagged KSHV proteins (mBFP2-ORF6, mCardinal-ORF52, and mCherry-LANA). Rainbow-KSHV replicated similarly to a prototype reporter-KSHV, KSHVr.219, and wild-type BAC16 virus. Live imaging revealed unsynchronized initiation of reactivation and KSHV replication with diverse kinetics between individual cells. Cell fractionation revealed temporal gene regulation, in which early lytic gene expression was terminated in late protein-expressing cells. Finally, isolation of fluorescence-positive cells from nonresponders increased dynamic ranges of downstream experiments 10-fold. Thus, this study demonstrates a tool to examine heterogenic responses of KSHV reactivation for a deeper understanding of KSHV replication.
    MeSH term(s) Cell Line ; Fluorescence ; Gene Expression Regulation, Viral/genetics ; Genes, Viral/genetics ; Herpesvirus 8, Human/genetics ; Herpesvirus 8, Human/physiology ; Host-Pathogen Interactions/genetics ; Host-Pathogen Interactions/physiology ; Humans ; Viral Proteins/genetics ; Virus Replication/genetics
    Chemical Substances Viral Proteins
    Language English
    Publishing date 2020-03-31
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 80174-4
    ISSN 1098-5514 ; 0022-538X
    ISSN (online) 1098-5514
    ISSN 0022-538X
    DOI 10.1128/JVI.01565-19
    Database MEDical Literature Analysis and Retrieval System OnLINE

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