Transcriptomic Analysis Reveals Novel Mechanistic Insight into Murine Biological Responses to Multi-Walled Carbon Nanotubes in Lungs and Cultured Lung Epithelial Cells

Sarah Søs Poulsen, Nicklas Raun Jacobsen, Sarah Labib, Dongmei Wu, Mainul Husain, Andrew Williams, Jesper Bøgelund, Ole Andersen, Carsten Købler, Kristian Mølhave, Zdenka Kyjovska, Anne Saber, Håkan Wallin, carole Yauk, Ulla Vogel, Sabina Halappanavar

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

    Resumé

    There is great interest in substituting animal work with in vitro experimentation in human health risk assessment; however, there are only few comparisons of in vitro and in vivo biological responses to engineered nanomaterials. We used high-content genomics tools to compare in vivo pulmonary responses of multiwalled carbon nanotubes (MWCNT) to those in vitro in cultured lung epithelial cells (FE1) at the global transcriptomic level. Primary size, surface area and other properties of MWCNT- XNRI -7 (Mitsui7) were characterized using DLS, SEM and TEM. Mice were exposed via a single intratracheal instillation to 18, 54, or 162 μg of Mitsui7/mouse. FE1 cells were incubated with 12.5, 25 and 100 μg/ml of Mitsui7. Tissue and cell samples were collected at 24 hours post-exposure. DNA microarrays were employed to establish mechanistic differences and similarities between the two models. Microarray results were confirmed using gene-specific RT-qPCR. Bronchoalveolar lavage (BAL) fluid was assessed for indications of inflammation in vivo. A strong dose-dependent activation of acute phase and inflammation response was observed in mouse lungs reflective mainly of an inflammatory response as observed in BAL. In vitro, a wide variety of core cellular functions were affected including transcription, cell cycle, and cellular growth and proliferation. Oxidative stress, fibrosis and inflammation processes were altered in both models. Although there were similarities observed between the two models at the pathway-level, the specific genes altered under these pathways were different, suggesting that the underlying mechanisms of responses are different in cells in culture and the lung tissue. Our results suggest that careful consideration should be given in selecting relevant endpoints when substituting animal with in vitro testing.
    OriginalsprogEngelsk
    TidsskriftP L o S One
    Vol/bind2013
    Sider (fra-til)1-25
    Antal sider25
    ISSN1932-6203
    DOI
    StatusUdgivet - 19 nov. 2013

    Citer dette

    Poulsen, Sarah Søs ; Raun Jacobsen, Nicklas ; Labib, Sarah ; Wu, Dongmei ; Husain, Mainul ; Williams, Andrew ; Bøgelund, Jesper ; Andersen, Ole ; Købler, Carsten ; Mølhave, Kristian ; Kyjovska, Zdenka ; Saber, Anne ; Wallin, Håkan ; Yauk, carole ; Vogel, Ulla ; Halappanavar, Sabina. / Transcriptomic Analysis Reveals Novel Mechanistic Insight into Murine Biological Responses to Multi-Walled Carbon Nanotubes in Lungs and Cultured Lung Epithelial Cells. I: P L o S One. 2013 ; Bind 2013. s. 1-25.
    @article{5ba24fc04faf4be6a51d3c83c957da83,
    title = "Transcriptomic Analysis Reveals Novel Mechanistic Insight into Murine Biological Responses to Multi-Walled Carbon Nanotubes in Lungs and Cultured Lung Epithelial Cells",
    abstract = "There is great interest in substituting animal work with in vitro experimentation in human health risk assessment; however, there are only few comparisons of in vitro and in vivo biological responses to engineered nanomaterials. We used high-content genomics tools to compare in vivo pulmonary responses of multiwalled carbon nanotubes (MWCNT) to those in vitro in cultured lung epithelial cells (FE1) at the global transcriptomic level. Primary size, surface area and other properties of MWCNT- XNRI -7 (Mitsui7) were characterized using DLS, SEM and TEM. Mice were exposed via a single intratracheal instillation to 18, 54, or 162 μg of Mitsui7/mouse. FE1 cells were incubated with 12.5, 25 and 100 μg/ml of Mitsui7. Tissue and cell samples were collected at 24 hours post-exposure. DNA microarrays were employed to establish mechanistic differences and similarities between the two models. Microarray results were confirmed using gene-specific RT-qPCR. Bronchoalveolar lavage (BAL) fluid was assessed for indications of inflammation in vivo. A strong dose-dependent activation of acute phase and inflammation response was observed in mouse lungs reflective mainly of an inflammatory response as observed in BAL. In vitro, a wide variety of core cellular functions were affected including transcription, cell cycle, and cellular growth and proliferation. Oxidative stress, fibrosis and inflammation processes were altered in both models. Although there were similarities observed between the two models at the pathway-level, the specific genes altered under these pathways were different, suggesting that the underlying mechanisms of responses are different in cells in culture and the lung tissue. Our results suggest that careful consideration should be given in selecting relevant endpoints when substituting animal with in vitro testing.",
    author = "Poulsen, {Sarah S{\o}s} and {Raun Jacobsen}, Nicklas and Sarah Labib and Dongmei Wu and Mainul Husain and Andrew Williams and Jesper B{\o}gelund and Ole Andersen and Carsten K{\o}bler and Kristian M{\o}lhave and Zdenka Kyjovska and Anne Saber and H{\aa}kan Wallin and carole Yauk and Ulla Vogel and Sabina Halappanavar",
    year = "2013",
    month = "11",
    day = "19",
    doi = "10.1371/journal.pone.0080452",
    language = "English",
    volume = "2013",
    pages = "1--25",
    journal = "P L o S One",
    issn = "1932-6203",
    publisher = "Public Library of Science",

    }

    Poulsen, SS, Raun Jacobsen, N, Labib, S, Wu, D, Husain, M, Williams, A, Bøgelund, J, Andersen, O, Købler, C, Mølhave, K, Kyjovska, Z, Saber, A, Wallin, H, Yauk, C, Vogel, U & Halappanavar, S 2013, 'Transcriptomic Analysis Reveals Novel Mechanistic Insight into Murine Biological Responses to Multi-Walled Carbon Nanotubes in Lungs and Cultured Lung Epithelial Cells', P L o S One, bind 2013, s. 1-25. https://doi.org/10.1371/journal.pone.0080452

    Transcriptomic Analysis Reveals Novel Mechanistic Insight into Murine Biological Responses to Multi-Walled Carbon Nanotubes in Lungs and Cultured Lung Epithelial Cells. / Poulsen, Sarah Søs; Raun Jacobsen, Nicklas; Labib, Sarah; Wu, Dongmei; Husain, Mainul ; Williams, Andrew; Bøgelund, Jesper; Andersen, Ole; Købler, Carsten; Mølhave, Kristian; Kyjovska, Zdenka; Saber, Anne ; Wallin, Håkan; Yauk, carole; Vogel, Ulla; Halappanavar, Sabina.

    I: P L o S One, Bind 2013, 19.11.2013, s. 1-25.

    Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

    TY - JOUR

    T1 - Transcriptomic Analysis Reveals Novel Mechanistic Insight into Murine Biological Responses to Multi-Walled Carbon Nanotubes in Lungs and Cultured Lung Epithelial Cells

    AU - Poulsen, Sarah Søs

    AU - Raun Jacobsen, Nicklas

    AU - Labib, Sarah

    AU - Wu, Dongmei

    AU - Husain, Mainul

    AU - Williams, Andrew

    AU - Bøgelund, Jesper

    AU - Andersen, Ole

    AU - Købler, Carsten

    AU - Mølhave, Kristian

    AU - Kyjovska, Zdenka

    AU - Saber, Anne

    AU - Wallin, Håkan

    AU - Yauk, carole

    AU - Vogel, Ulla

    AU - Halappanavar, Sabina

    PY - 2013/11/19

    Y1 - 2013/11/19

    N2 - There is great interest in substituting animal work with in vitro experimentation in human health risk assessment; however, there are only few comparisons of in vitro and in vivo biological responses to engineered nanomaterials. We used high-content genomics tools to compare in vivo pulmonary responses of multiwalled carbon nanotubes (MWCNT) to those in vitro in cultured lung epithelial cells (FE1) at the global transcriptomic level. Primary size, surface area and other properties of MWCNT- XNRI -7 (Mitsui7) were characterized using DLS, SEM and TEM. Mice were exposed via a single intratracheal instillation to 18, 54, or 162 μg of Mitsui7/mouse. FE1 cells were incubated with 12.5, 25 and 100 μg/ml of Mitsui7. Tissue and cell samples were collected at 24 hours post-exposure. DNA microarrays were employed to establish mechanistic differences and similarities between the two models. Microarray results were confirmed using gene-specific RT-qPCR. Bronchoalveolar lavage (BAL) fluid was assessed for indications of inflammation in vivo. A strong dose-dependent activation of acute phase and inflammation response was observed in mouse lungs reflective mainly of an inflammatory response as observed in BAL. In vitro, a wide variety of core cellular functions were affected including transcription, cell cycle, and cellular growth and proliferation. Oxidative stress, fibrosis and inflammation processes were altered in both models. Although there were similarities observed between the two models at the pathway-level, the specific genes altered under these pathways were different, suggesting that the underlying mechanisms of responses are different in cells in culture and the lung tissue. Our results suggest that careful consideration should be given in selecting relevant endpoints when substituting animal with in vitro testing.

    AB - There is great interest in substituting animal work with in vitro experimentation in human health risk assessment; however, there are only few comparisons of in vitro and in vivo biological responses to engineered nanomaterials. We used high-content genomics tools to compare in vivo pulmonary responses of multiwalled carbon nanotubes (MWCNT) to those in vitro in cultured lung epithelial cells (FE1) at the global transcriptomic level. Primary size, surface area and other properties of MWCNT- XNRI -7 (Mitsui7) were characterized using DLS, SEM and TEM. Mice were exposed via a single intratracheal instillation to 18, 54, or 162 μg of Mitsui7/mouse. FE1 cells were incubated with 12.5, 25 and 100 μg/ml of Mitsui7. Tissue and cell samples were collected at 24 hours post-exposure. DNA microarrays were employed to establish mechanistic differences and similarities between the two models. Microarray results were confirmed using gene-specific RT-qPCR. Bronchoalveolar lavage (BAL) fluid was assessed for indications of inflammation in vivo. A strong dose-dependent activation of acute phase and inflammation response was observed in mouse lungs reflective mainly of an inflammatory response as observed in BAL. In vitro, a wide variety of core cellular functions were affected including transcription, cell cycle, and cellular growth and proliferation. Oxidative stress, fibrosis and inflammation processes were altered in both models. Although there were similarities observed between the two models at the pathway-level, the specific genes altered under these pathways were different, suggesting that the underlying mechanisms of responses are different in cells in culture and the lung tissue. Our results suggest that careful consideration should be given in selecting relevant endpoints when substituting animal with in vitro testing.

    U2 - 10.1371/journal.pone.0080452

    DO - 10.1371/journal.pone.0080452

    M3 - Journal article

    VL - 2013

    SP - 1

    EP - 25

    JO - P L o S One

    JF - P L o S One

    SN - 1932-6203

    ER -