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  1. Article ; Online: The Hippo Pathway, YAP/TAZ, and the Plasma Membrane.

    Rausch, Valentina / Hansen, Carsten G

    Trends in cell biology

    2019  Volume 30, Issue 1, Page(s) 32–48

    Abstract: The plasma membrane allows the cell to sense and adapt to changes in the extracellular environment by relaying external inputs via intracellular signaling networks. One central cellular signaling pathway is the Hippo pathway, which regulates homeostasis ... ...

    Abstract The plasma membrane allows the cell to sense and adapt to changes in the extracellular environment by relaying external inputs via intracellular signaling networks. One central cellular signaling pathway is the Hippo pathway, which regulates homeostasis and plays chief roles in carcinogenesis and regenerative processes. Recent studies have found that mechanical stimuli and diffusible chemical components can regulate the Hippo pathway primarily through receptors embedded in the plasma membrane. Morphologically defined structures within the plasma membrane, such as cellular junctions, focal adhesions, primary cilia, caveolae, clathrin-coated pits, and plaques play additional key roles. Here, we discuss recent evidence highlighting the importance of these specialized plasma membrane domains in cellular feedback via the Hippo pathway.
    MeSH term(s) Adaptor Proteins, Signal Transducing/metabolism ; Animals ; Cell Membrane/metabolism ; Cilia/metabolism ; Humans ; Protein-Serine-Threonine Kinases/metabolism ; Signal Transduction ; Transcription Factors/metabolism
    Chemical Substances Adaptor Proteins, Signal Transducing ; Transcription Factors ; Protein-Serine-Threonine Kinases (EC 2.7.11.1)
    Language English
    Publishing date 2019-12-02
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 30122-x
    ISSN 1879-3088 ; 0962-8924
    ISSN (online) 1879-3088
    ISSN 0962-8924
    DOI 10.1016/j.tcb.2019.10.005
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  2. Article: The Hippo Pathway in Prostate Cancer.

    Salem, Omar / Hansen, Carsten G

    Cells

    2019  Volume 8, Issue 4

    Abstract: Despite recent efforts, prostate cancer (PCa) remains one of the most common cancers in men. Currently, there is no effective treatment for castration-resistant prostate cancer (CRPC). There is, therefore, an urgent need to identify new therapeutic ... ...

    Abstract Despite recent efforts, prostate cancer (PCa) remains one of the most common cancers in men. Currently, there is no effective treatment for castration-resistant prostate cancer (CRPC). There is, therefore, an urgent need to identify new therapeutic targets. The Hippo pathway and its downstream effectors-the transcriptional co-activators, Yes-associated protein (YAP) and its paralog, transcriptional co-activator with PDZ-binding motif (TAZ)-are foremost regulators of stem cells and cancer biology. Defective Hippo pathway signaling and YAP/TAZ hyperactivation are common across various cancers. Here, we draw on insights learned from other types of cancers and review the latest advances linking the Hippo pathway and YAP/TAZ to PCa onset and progression. We examine the regulatory interaction between Hippo-YAP/TAZ and the androgen receptor (AR), as main regulators of PCa development, and how uncontrolled expression of YAP/TAZ drives castration resistance by inducing cellular stemness. Finally, we survey the potential therapeutic targeting of the Hippo pathway and YAP/TAZ to overcome PCa.
    MeSH term(s) Adaptor Proteins, Signal Transducing/metabolism ; Humans ; Intracellular Signaling Peptides and Proteins/metabolism ; Male ; Phosphoproteins ; Prostatic Neoplasms/metabolism ; Prostatic Neoplasms/physiopathology ; Protein-Serine-Threonine Kinases/metabolism ; Protein-Serine-Threonine Kinases/physiology ; Signal Transduction ; Trans-Activators/metabolism ; Transcription Factors/metabolism
    Chemical Substances Adaptor Proteins, Signal Transducing ; Intracellular Signaling Peptides and Proteins ; Phosphoproteins ; Trans-Activators ; Transcription Factors ; WWTR1 protein, human ; YAP1 protein, human ; Hippo protein, human (EC 2.7.11.1) ; Protein-Serine-Threonine Kinases (EC 2.7.11.1)
    Language English
    Publishing date 2019-04-23
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Review
    ZDB-ID 2661518-6
    ISSN 2073-4409
    ISSN 2073-4409
    DOI 10.3390/cells8040370
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  3. Article ; Online: The Hippo Pathway in Prostate Cancer

    Omar Salem / Carsten G. Hansen

    Cells, Vol 8, Iss 4, p

    2019  Volume 370

    Abstract: Despite recent efforts, prostate cancer (PCa) remains one of the most common cancers in men. Currently, there is no effective treatment for castration-resistant prostate cancer (CRPC). There is, therefore, an urgent need to identify new therapeutic ... ...

    Abstract Despite recent efforts, prostate cancer (PCa) remains one of the most common cancers in men. Currently, there is no effective treatment for castration-resistant prostate cancer (CRPC). There is, therefore, an urgent need to identify new therapeutic targets. The Hippo pathway and its downstream effectors—the transcriptional co-activators, Yes-associated protein (YAP) and its paralog, transcriptional co-activator with PDZ-binding motif (TAZ)—are foremost regulators of stem cells and cancer biology. Defective Hippo pathway signaling and YAP/TAZ hyperactivation are common across various cancers. Here, we draw on insights learned from other types of cancers and review the latest advances linking the Hippo pathway and YAP/TAZ to PCa onset and progression. We examine the regulatory interaction between Hippo-YAP/TAZ and the androgen receptor (AR), as main regulators of PCa development, and how uncontrolled expression of YAP/TAZ drives castration resistance by inducing cellular stemness. Finally, we survey the potential therapeutic targeting of the Hippo pathway and YAP/TAZ to overcome PCa.
    Keywords hippo pathway ; YAP/TAZ ; prostate cancer ; castration resistance ; signal cross-talk ; feedback loops ; Biology (General) ; QH301-705.5
    Subject code 570
    Language English
    Publishing date 2019-04-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
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  4. Article ; Online: Immunofluorescence Study of Endogenous YAP in Mammalian Cells.

    Rausch, Valentina / Hansen, Carsten G

    Methods in molecular biology (Clifton, N.J.)

    2018  Volume 1893, Page(s) 97–106

    Abstract: Immunocytochemistry enables determination of cellular localization and relative abundance of proteins. This protocol describes a rapid and cost-effective approach to study the cellular localization of YAP (and TAZ), the transcriptional co activators of ... ...

    Abstract Immunocytochemistry enables determination of cellular localization and relative abundance of proteins. This protocol describes a rapid and cost-effective approach to study the cellular localization of YAP (and TAZ), the transcriptional co activators of the Hippo pathway, in mammalian cells. Cells are seeded onto coated cover slips, cultivated and treated as required. Subsequently, they are chemically fixed, and cellular proteins are fluorescently labeled by means of specific antibodies. Multiplexing antibodies enables ascertaining the subcellular localization of YAP and TAZ and thereby also the activation state of the Hippo pathway in various cell types.
    MeSH term(s) Animals ; Cell Culture Techniques ; Cell Line ; Fluorescent Antibody Technique ; Humans ; Microscopy ; Nuclear Proteins/metabolism ; Signal Transduction ; Transcription Factors/metabolism
    Chemical Substances Nuclear Proteins ; Transcription Factors ; YY1AP1 protein, human
    Language English
    Publishing date 2018-12-18
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-4939-8910-2_8
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  5. Article ; Online: The Hippo pathway drives the cellular response to hydrostatic pressure.

    Park, Jiwon / Jia, Siyang / Salter, Donald / Bagnaninchi, Pierre / Hansen, Carsten G

    The EMBO journal

    2022  Volume 41, Issue 13, Page(s) e108719

    Abstract: Cells need to rapidly and precisely react to multiple mechanical and chemical stimuli in order to ensure precise context-dependent responses. This requires dynamic cellular signalling events that ensure homeostasis and plasticity when needed. A less well- ...

    Abstract Cells need to rapidly and precisely react to multiple mechanical and chemical stimuli in order to ensure precise context-dependent responses. This requires dynamic cellular signalling events that ensure homeostasis and plasticity when needed. A less well-understood process is cellular response to elevated interstitial fluid pressure, where the cell senses and responds to changes in extracellular hydrostatic pressure. Here, using quantitative label-free digital holographic imaging, combined with genome editing, biochemical assays and confocal imaging, we analyse the temporal cellular response to hydrostatic pressure. Upon elevated cyclic hydrostatic pressure, the cell responds by rapid, dramatic and reversible changes in cellular volume. We show that YAP and TAZ, the co-transcriptional regulators of the Hippo signalling pathway, control cell volume and that cells without YAP and TAZ have lower plasma membrane tension. We present direct evidence that YAP/TAZ drive the cellular response to hydrostatic pressure, a process that is at least partly mediated via clathrin-dependent endocytosis. Additionally, upon elevated oscillating hydrostatic pressure, YAP/TAZ are activated and induce TEAD-mediated transcription and expression of cellular components involved in dynamic regulation of cell volume and extracellular matrix. This cellular response confers a feedback loop that allows the cell to robustly respond to changes in interstitial fluid pressure.
    MeSH term(s) Hippo Signaling Pathway ; Homeostasis ; Hydrostatic Pressure ; Phosphoproteins/metabolism ; Protein Serine-Threonine Kinases/genetics ; Transcription Factors/genetics ; Transcription Factors/metabolism
    Chemical Substances Phosphoproteins ; Transcription Factors ; Protein Serine-Threonine Kinases (EC 2.7.11.1)
    Language English
    Publishing date 2022-06-15
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 586044-1
    ISSN 1460-2075 ; 0261-4189
    ISSN (online) 1460-2075
    ISSN 0261-4189
    DOI 10.15252/embj.2021108719
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  6. Article ; Online: The LKB1-TSSK1B axis controls YAP phosphorylation to regulate the Hippo-YAP pathway.

    Kim, Cho-Long / Lim, Su-Bin / Choi, Sue-Hee / Kim, Dong Hyun / Sim, Ye Eun / Jo, Eun-Hye / Kim, Keeeun / Lee, Keesook / Park, Hee-Sae / Lim, Su Bin / Kang, Li-Jung / Jeong, Han-Sol / Lee, Youngsoo / Hansen, Carsten G / Mo, Jung-Soon

    Cell death & disease

    2024  Volume 15, Issue 1, Page(s) 76

    Abstract: The Hippo pathway's main effector, Yes-associated protein (YAP), plays a crucial role in tumorigenesis as a transcriptional coactivator. YAP's phosphorylation by core upstream components of the Hippo pathway, such as mammalian Ste20 kinase 1/2 (MST1/2), ... ...

    Abstract The Hippo pathway's main effector, Yes-associated protein (YAP), plays a crucial role in tumorigenesis as a transcriptional coactivator. YAP's phosphorylation by core upstream components of the Hippo pathway, such as mammalian Ste20 kinase 1/2 (MST1/2), mitogen-activated protein kinase kinase kinase kinases (MAP4Ks), and their substrate, large tumor suppressor 1/2 (LATS1/2), influences YAP's subcellular localization, stability, and transcriptional activity. However, recent research suggests the existence of alternative pathways that phosphorylate YAP, independent of these core upstream Hippo pathway components, raising questions about additional means to inactivate YAP. In this study, we present evidence demonstrating that TSSK1B, a calcium/calmodulin-dependent protein kinase (CAMK) superfamily member, is a negative regulator of YAP, suppressing cellular proliferation and oncogenic transformation. Mechanistically, TSSK1B inhibits YAP through two distinct pathways. Firstly, the LKB1-TSSK1B axis directly phosphorylates YAP at Ser94, inhibiting the YAP-TEAD complex's formation and suppressing its target genes' expression. Secondly, the TSSK1B-LATS1/2 axis inhibits YAP via phosphorylation at Ser127. Our findings reveal the involvement of TSSK1B-mediated molecular mechanisms in the Hippo-YAP pathway, emphasizing the importance of multilevel regulation in critical cellular decision-making processes.
    MeSH term(s) Animals ; Humans ; Phosphorylation ; Hippo Signaling Pathway ; Signal Transduction ; YAP-Signaling Proteins ; Protein Serine-Threonine Kinases/genetics ; Protein Serine-Threonine Kinases/metabolism ; Cell Transformation, Neoplastic/metabolism ; Cell Proliferation/physiology ; Phosphoproteins/metabolism ; Mammals
    Chemical Substances YAP-Signaling Proteins ; Protein Serine-Threonine Kinases (EC 2.7.11.1) ; Phosphoproteins
    Language English
    Publishing date 2024-01-20
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2541626-1
    ISSN 2041-4889 ; 2041-4889
    ISSN (online) 2041-4889
    ISSN 2041-4889
    DOI 10.1038/s41419-024-06465-4
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  7. Article ; Online: Immune complex-induced apoptosis and concurrent immune complex clearance are anti-inflammatory neutrophil functions.

    Karmakar, Utsa / Chu, Julia Y / Sundaram, Kruthika / Astier, Anne L / Garside, Hannah / Hansen, Carsten G / Dransfield, Ian / Vermeren, Sonja

    Cell death & disease

    2021  Volume 12, Issue 4, Page(s) 296

    Abstract: Persistent neutrophilic inflammation drives host damage in autoimmune diseases that are characterized by abundant immune complexes. Insoluble immune complexes (iICs) potently activate pro-inflammatory neutrophil effector functions. We and others have ... ...

    Abstract Persistent neutrophilic inflammation drives host damage in autoimmune diseases that are characterized by abundant immune complexes. Insoluble immune complexes (iICs) potently activate pro-inflammatory neutrophil effector functions. We and others have shown that iICs also promote resolution of inflammation via stimulation of neutrophil apoptosis. We demonstrate here that iICs trigger FcγRIIa-dependent neutrophil macropinocytosis, leading to the rapid uptake, and subsequent degradation of iICs. We provide evidence that concurrent iIC-induced neutrophil apoptosis is distinct from phagocytosis-induced cell death. First, uptake of iICs occurs by FcγRII-stimulated macropinocytosis, rather than phagocytosis. Second, production of reactive oxygen species, but not iIC-internalization is a pre-requisite for iIC-induced neutrophil apoptosis. Our findings identify a previously unknown mechanism by which neutrophils can remove pro-inflammatory iICs from the circulation. Together iIC clearance and iIC-induced neutrophil apoptosis may act to prevent the potential escalation of neutrophilic inflammation in response to iICs.
    MeSH term(s) Antigen-Antibody Complex/metabolism ; Apoptosis ; Humans ; Inflammation/immunology ; Neutrophils/immunology
    Chemical Substances Antigen-Antibody Complex
    Language English
    Publishing date 2021-03-19
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2541626-1
    ISSN 2041-4889 ; 2041-4889
    ISSN (online) 2041-4889
    ISSN 2041-4889
    DOI 10.1038/s41419-021-03528-8
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  8. Article ; Online: Delineation uncertainties of tumour volumes on MRI of head and neck cancer patients.

    Zukauskaite, Ruta / Rumley, Christopher N / Hansen, Christian R / Jameson, Michael G / Trada, Yuvnik / Johansen, Jørgen / Gyldenkerne, Niels / Eriksen, Jesper G / Aly, Farhannah / Christensen, Rasmus L / Lee, Mark / Brink, Carsten / Holloway, Lois

    Clinical and translational radiation oncology

    2022  Volume 36, Page(s) 121–126

    Abstract: Background: During the last decade, radiotherapy using MR Linac has gone from research to clinical implementation for different cancer locations. For head and neck cancer (HNC), target delineation based only on MR images is not yet standard, and the ... ...

    Abstract Background: During the last decade, radiotherapy using MR Linac has gone from research to clinical implementation for different cancer locations. For head and neck cancer (HNC), target delineation based only on MR images is not yet standard, and the utilisation of MRI instead of PET/CT in radiotherapy planning is not well established. We aimed to analyse the inter-observer variation (IOV) in delineating GTV (gross tumour volume) on MR images only for patients with HNC.
    Material/methods: 32 HNC patients from two independent departments were included. Four clinical oncologists from Denmark and four radiation oncologists from Australia had independently contoured primary tumour GTVs (GTV-T) and nodal GTVs (GTV-N) on T2-weighted MR images obtained at the time of treatment planning. Observers were provided with sets of images, delineation guidelines and patient synopsis. Simultaneous truth and performance level estimation (STAPLE) reference volumes were generated for each structure using all observer contours. The IOV was assessed using the DICE Similarity Coefficient (DSC) and mean absolute surface distance (MASD).
    Results: 32 GTV-Ts and 68 GTV-Ns were contoured per observer. The median MASD for GTV-Ts and GTV-Ns across all patients was 0.17 cm (range 0.08-0.39 cm) and 0.07 cm (range 0.04-0.33 cm), respectively. Median DSC relative to a STAPLE volume for GTV-Ts and GTV-Ns across all patients were 0.73 and 0.76, respectively. A significant correlation was seen between median DSCs and median volumes of GTV-Ts (Spearman correlation coefficient 0.76, p < 0.001) and of GTV-Ns (Spearman correlation coefficient 0.55, p < 0.001).
    Conclusion: Contouring GTVs in patients with HNC on MRI showed that the median IOV for GTV-T and GTV-N was below 2 mm, based on observes from two separate radiation departments. However, there are still specific regions in tumours that are difficult to resolve as either malignant tissue or oedema that potentially could be improved by further training in MR-only delineation.
    Language English
    Publishing date 2022-08-06
    Publishing country Ireland
    Document type Journal Article
    ISSN 2405-6308
    ISSN (online) 2405-6308
    DOI 10.1016/j.ctro.2022.08.005
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  9. Article ; Online: Delineation uncertainties of tumour volumes on MRI of head and neck cancer patients

    Ruta Zukauskaite / Christopher N. Rumley / Christian R. Hansen / Michael G. Jameson / Yuvnik Trada / Jørgen Johansen / Niels Gyldenkerne / Jesper G. Eriksen / Farhannah Aly / Rasmus L. Christensen / Mark Lee / Carsten Brink / Lois Holloway

    Clinical and Translational Radiation Oncology, Vol 36, Iss , Pp 121-

    2022  Volume 126

    Abstract: Background: During the last decade, radiotherapy using MR Linac has gone from research to clinical implementation for different cancer locations. For head and neck cancer (HNC), target delineation based only on MR images is not yet standard, and the ... ...

    Abstract Background: During the last decade, radiotherapy using MR Linac has gone from research to clinical implementation for different cancer locations. For head and neck cancer (HNC), target delineation based only on MR images is not yet standard, and the utilisation of MRI instead of PET/CT in radiotherapy planning is not well established. We aimed to analyse the inter-observer variation (IOV) in delineating GTV (gross tumour volume) on MR images only for patients with HNC. Material/methods: 32 HNC patients from two independent departments were included. Four clinical oncologists from Denmark and four radiation oncologists from Australia had independently contoured primary tumour GTVs (GTV-T) and nodal GTVs (GTV-N) on T2-weighted MR images obtained at the time of treatment planning. Observers were provided with sets of images, delineation guidelines and patient synopsis. Simultaneous truth and performance level estimation (STAPLE) reference volumes were generated for each structure using all observer contours. The IOV was assessed using the DICE Similarity Coefficient (DSC) and mean absolute surface distance (MASD). Results: 32 GTV-Ts and 68 GTV-Ns were contoured per observer. The median MASD for GTV-Ts and GTV-Ns across all patients was 0.17 cm (range 0.08–0.39 cm) and 0.07 cm (range 0.04–0.33 cm), respectively. Median DSC relative to a STAPLE volume for GTV-Ts and GTV-Ns across all patients were 0.73 and 0.76, respectively. A significant correlation was seen between median DSCs and median volumes of GTV-Ts (Spearman correlation coefficient 0.76, p < 0.001) and of GTV-Ns (Spearman correlation coefficient 0.55, p < 0.001). Conclusion: Contouring GTVs in patients with HNC on MRI showed that the median IOV for GTV-T and GTV-N was below 2 mm, based on observes from two separate radiation departments. However, there are still specific regions in tumours that are difficult to resolve as either malignant tissue or oedema that potentially could be improved by further training in MR-only delineation.
    Keywords Inter-observer variation ; Target delineation ; Head and neck cancer ; MRI ; Medical physics. Medical radiology. Nuclear medicine ; R895-920 ; Neoplasms. Tumors. Oncology. Including cancer and carcinogens ; RC254-282
    Subject code 610
    Language English
    Publishing date 2022-09-01T00:00:00Z
    Publisher Elsevier
    Document type Article ; Online
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  10. Article ; Online: Label2label: training a neural network to selectively restore cellular structures in fluorescence microscopy.

    Kölln, Lisa Sophie / Salem, Omar / Valli, Jessica / Hansen, Carsten Gram / McConnell, Gail

    Journal of cell science

    2022  Volume 135, Issue 3

    Abstract: Immunofluorescence microscopy is routinely used to visualise the spatial distribution of proteins that dictates their cellular function. However, unspecific antibody binding often results in high cytosolic background signals, decreasing the image ... ...

    Abstract Immunofluorescence microscopy is routinely used to visualise the spatial distribution of proteins that dictates their cellular function. However, unspecific antibody binding often results in high cytosolic background signals, decreasing the image contrast of a target structure. Recently, convolutional neural networks (CNNs) were successfully employed for image restoration in immunofluorescence microscopy, but current methods cannot correct for those background signals. We report a new method that trains a CNN to reduce unspecific signals in immunofluorescence images; we name this method label2label (L2L). In L2L, a CNN is trained with image pairs of two non-identical labels that target the same cellular structure. We show that after L2L training a network predicts images with significantly increased contrast of a target structure, which is further improved after implementing a multiscale structural similarity loss function. Here, our results suggest that sample differences in the training data decrease hallucination effects that are observed with other methods. We further assess the performance of a cycle generative adversarial network, and show that a CNN can be trained to separate structures in superposed immunofluorescence images of two targets.
    MeSH term(s) Cellular Structures ; Image Processing, Computer-Assisted/methods ; Microscopy, Fluorescence ; Neural Networks, Computer
    Language English
    Publishing date 2022-02-10
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2993-2
    ISSN 1477-9137 ; 0021-9533
    ISSN (online) 1477-9137
    ISSN 0021-9533
    DOI 10.1242/jcs.258994
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