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  1. Article ; Online: Implantation of engineered human microvasculature to study human infectious diseases in mouse models.

    Schönherr-Hellec, Sophia / Chatzopoulou, Eirini / Barnier, Jean-Philippe / Atlas, Yoann / Dupichaud, Sébastien / Guilbert, Thomas / Dupraz, Yves / Meyer, Julie / Chaussain, Catherine / Gorin, Caroline / Nassif, Xavier / Germain, Stephane / Muller, Laurent / Coureuil, Mathieu

    iScience

    2023  Volume 26, Issue 4, Page(s) 106286

    Abstract: Animal models for studying human pathogens are crucially lacking. We describe the implantation in mice of engineered human mature microvasculature consisting of endothelial and perivascular cells embedded in collagen hydrogel that allows investigation of ...

    Abstract Animal models for studying human pathogens are crucially lacking. We describe the implantation in mice of engineered human mature microvasculature consisting of endothelial and perivascular cells embedded in collagen hydrogel that allows investigation of pathogen interactions with the endothelium, including
    Language English
    Publishing date 2023-02-27
    Publishing country United States
    Document type Journal Article
    ISSN 2589-0042
    ISSN (online) 2589-0042
    DOI 10.1016/j.isci.2023.106286
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article: 2d and 3d human induced pluripotent stem cell-based models to dissect primary cilium involvement during neocortical development

    Boutaud, Lucile / Michael, Marie / Banal, Céline / Calderon, Damelys / Farcy, Sarah / Pernelle, Julie / Goudin, Nicolas / Maillard, Camille / Dimartino, Clémantine / Deleschaux, Cécile / Dupichaud, Sébastien / Lebreton, Corinne / Saunier, Sophie / Attié-Bitach, Tania / Bahi-Buisson, Nadia / Lefort, Nathalie / Thomas, Sophie

    Journal of visualized experiments. 2022 Mar. 25, , no. 181

    2022  

    Abstract: Primary cilia (PC) are non-motile dynamic microtubule-based organelles that protrude from the surface of most mammalian cells. They emerge from the older centriole during the G1/G0 phase of the cell cycle, while they disassemble as the cells re-enter the ...

    Abstract Primary cilia (PC) are non-motile dynamic microtubule-based organelles that protrude from the surface of most mammalian cells. They emerge from the older centriole during the G1/G0 phase of the cell cycle, while they disassemble as the cells re-enter the cell cycle at the G2/M phase boundary. They function as signal hubs, by detecting and transducing extracellular signals crucial for many cell processes. Similar to most cell types, all neocortical neural stem and progenitor cells (NSPCs) have been shown harboring a PC allowing them to sense and transduce specific signals required for the normal cerebral cortical development. Here, we provide detailed protocols to generate and characterize two-dimensional (2D) and three-dimensional (3D) cell-based models from human induced pluripotent stem cells (hIPSCs) to further dissect the involvement of PC during neocortical development. In particular, we present protocols to study the PC biogenesis and function in 2D neural rosette-derived NSPCs including the transduction of the Sonic Hedgehog (SHH) pathway. To take advantage of the three-dimensional (3D) organization of cerebral organoids, we describe a simple method for 3D imaging of in toto immunostained cerebral organoids. After optical clearing, rapid acquisition of entire organoids allows detection of both centrosomes and PC on neocortical progenitors and neurons of the whole organoid. Finally, we detail the procedure for immunostaining and clearing of thick free-floating organoid sections preserving a significant degree of 3D spatial information and allowing for the high-resolution acquisition required for the detailed qualitative and quantitative analysis of PC biogenesis and function.
    Keywords biogenesis ; centrioles ; humans ; organoids ; quantitative analysis ; spatial data
    Language English
    Dates of publication 2022-0325
    Size p. e62667.
    Publishing place Journal of Visualized Experiments
    Document type Article
    ZDB-ID 2259946-0
    ISSN 1940-087X
    ISSN 1940-087X
    DOI 10.3791/62667
    Database NAL-Catalogue (AGRICOLA)

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  3. Article ; Online: 2D and 3D Human Induced Pluripotent Stem Cell-Based Models to Dissect Primary Cilium Involvement during Neocortical Development.

    Boutaud, Lucile / Michael, Marie / Banal, Céline / Calderon, Damelys / Farcy, Sarah / Pernelle, Julie / Goudin, Nicolas / Maillard, Camille / Dimartino, Clémantine / Deleschaux, Cécile / Dupichaud, Sébastien / Lebreton, Corinne / Saunier, Sophie / Attié-Bitach, Tania / Bahi-Buisson, Nadia / Lefort, Nathalie / Thomas, Sophie

    Journal of visualized experiments : JoVE

    2022  , Issue 181

    Abstract: Primary cilia (PC) are non-motile dynamic microtubule-based organelles that protrude from the surface of most mammalian cells. They emerge from the older centriole during the G1/G0 phase of the cell cycle, while they disassemble as the cells re-enter the ...

    Abstract Primary cilia (PC) are non-motile dynamic microtubule-based organelles that protrude from the surface of most mammalian cells. They emerge from the older centriole during the G1/G0 phase of the cell cycle, while they disassemble as the cells re-enter the cell cycle at the G2/M phase boundary. They function as signal hubs, by detecting and transducing extracellular signals crucial for many cell processes. Similar to most cell types, all neocortical neural stem and progenitor cells (NSPCs) have been shown harboring a PC allowing them to sense and transduce specific signals required for the normal cerebral cortical development. Here, we provide detailed protocols to generate and characterize two-dimensional (2D) and three-dimensional (3D) cell-based models from human induced pluripotent stem cells (hIPSCs) to further dissect the involvement of PC during neocortical development. In particular, we present protocols to study the PC biogenesis and function in 2D neural rosette-derived NSPCs including the transduction of the Sonic Hedgehog (SHH) pathway. To take advantage of the three-dimensional (3D) organization of cerebral organoids, we describe a simple method for 3D imaging of in toto immunostained cerebral organoids. After optical clearing, rapid acquisition of entire organoids allows detection of both centrosomes and PC on neocortical progenitors and neurons of the whole organoid. Finally, we detail the procedure for immunostaining and clearing of thick free-floating organoid sections preserving a significant degree of 3D spatial information and allowing for the high-resolution acquisition required for the detailed qualitative and quantitative analysis of PC biogenesis and function.
    MeSH term(s) Animals ; Cell Differentiation/physiology ; Cilia/metabolism ; Hedgehog Proteins/metabolism ; Humans ; Induced Pluripotent Stem Cells ; Mammals/metabolism ; Neocortex ; Organoids/metabolism
    Chemical Substances Hedgehog Proteins
    Language English
    Publishing date 2022-03-25
    Publishing country United States
    Document type Journal Article ; Video-Audio Media ; Research Support, Non-U.S. Gov't
    ZDB-ID 2259946-0
    ISSN 1940-087X ; 1940-087X
    ISSN (online) 1940-087X
    ISSN 1940-087X
    DOI 10.3791/62667
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Activation of cGAS/STING pathway upon paramyxovirus infection.

    Iampietro, Mathieu / Dumont, Claire / Mathieu, Cyrille / Spanier, Julia / Robert, Jonathan / Charpenay, Aude / Dupichaud, Sébastien / Dhondt, Kévin P / Aurine, Noémie / Pelissier, Rodolphe / Ferren, Marion / Mély, Stéphane / Gerlier, Denis / Kalinke, Ulrich / Horvat, Branka

    iScience

    2021  Volume 24, Issue 6, Page(s) 102519

    Abstract: During inflammatory diseases, cancer, and infection, the cGAS/STING pathway is known to recognize foreign or self-DNA in the cytosol and activate an innate immune response. Here, we report that negative-strand RNA paramyxoviruses, Nipah virus (NiV), and ... ...

    Abstract During inflammatory diseases, cancer, and infection, the cGAS/STING pathway is known to recognize foreign or self-DNA in the cytosol and activate an innate immune response. Here, we report that negative-strand RNA paramyxoviruses, Nipah virus (NiV), and measles virus (MeV), can also trigger the cGAS/STING axis. Although mice deficient for MyD88, TRIF, and MAVS still moderately control NiV infection when compared with wild-type mice, additional STING deficiency resulted in 100% lethality, suggesting synergistic roles of these pathways in host protection. Moreover, deletion of cGAS or STING resulted in decreased type I interferon production with enhanced paramyxoviral infection in both human and murine cells. Finally, the phosphorylation and ubiquitination of STING, observed during viral infections, confirmed the activation of cGAS/STING pathway by NiV and MeV. Our data suggest that cGAS/STING activation is critical in controlling paramyxovirus infection and possibly represents attractive targets to develop countermeasures against severe disease induced by these pathogens.
    Language English
    Publishing date 2021-05-07
    Publishing country United States
    Document type Journal Article
    ISSN 2589-0042
    ISSN (online) 2589-0042
    DOI 10.1016/j.isci.2021.102519
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: DOCK11 deficiency in patients with X-linked actinopathy and autoimmunity.

    Boussard, Charlotte / Delage, Laure / Gajardo, Tania / Kauskot, Alexandre / Batignes, Maxime / Goudin, Nicolas / Stolzenberg, Marie-Claude / Brunaud, Camille / Panikulam, Patricia / Riller, Quentin / Moya-Nilges, Maryse / Solarz, Jean / Repérant, Christelle / Durel, Béatrice / Bordet, Jean-Claude / Pellé, Olivier / Lebreton, Corinne / Magérus, Aude / Pirabakaran, Vithura /
    Vargas, Pablo / Dupichaud, Sébastien / Jeanpierre, Marie / Vinit, Angélique / Zarhrate, Mohammed / Masson, Cécile / Aladjidi, Nathalie / Arkwright, Peter D / Bader-Meunier, Brigitte / Baron Joly, Sandrine / Benadiba, Joy / Bernard, Elise / Berrebi, Dominique / Bodemer, Christine / Castelle, Martin / Charbit-Henrion, Fabienne / Chbihi, Marwa / Debray, Agathe / Drabent, Philippe / Fraitag, Sylvie / Hié, Miguel / Landman-Parker, Judith / Lhermitte, Ludovic / Moshous, Despina / Rohrlich, Pierre / Ruemmele, Frank / Welfringer-Morin, Anne / Tusseau, Maud / Belot, Alexandre / Cerf-Bensussan, Nadine / Roelens, Marie / Picard, Capucine / Neven, Bénédicte / Fischer, Alain / Callebaut, Isabelle / Ménager, Mickaël / Sepulveda, Fernando E / Adam, Frédéric / Rieux-Laucat, Frédéric

    Blood

    2023  Volume 141, Issue 22, Page(s) 2713–2726

    Abstract: Dedicator of cytokinesis (DOCK) proteins play a central role in actin cytoskeleton regulation. This is highlighted by the DOCK2 and DOCK8 deficiencies leading to actinopathies and immune deficiencies. DOCK8 and DOCK11 activate CDC42, a Rho-guanosine ... ...

    Abstract Dedicator of cytokinesis (DOCK) proteins play a central role in actin cytoskeleton regulation. This is highlighted by the DOCK2 and DOCK8 deficiencies leading to actinopathies and immune deficiencies. DOCK8 and DOCK11 activate CDC42, a Rho-guanosine triphosphate hydrolases involved in actin cytoskeleton dynamics, among many cellular functions. The role of DOCK11 in human immune disease has been long suspected but, to the best of our knowledge, has never been described to date. We studied 8 male patients, from 7 unrelated families, with hemizygous DOCK11 missense variants leading to reduced DOCK11 expression. The patients were presenting with early-onset autoimmunity, including cytopenia, systemic lupus erythematosus, skin, and digestive manifestations. Patients' platelets exhibited abnormal ultrastructural morphology and spreading as well as impaired CDC42 activity. In vitro activated T cells and B-lymphoblastoid cell lines from patients exhibited aberrant protrusions and abnormal migration speed in confined channels concomitant with altered actin polymerization during migration. Knock down of DOCK11 recapitulated these abnormal cellular phenotypes in monocytes-derived dendritic cells and primary activated T cells from healthy controls. Lastly, in line with the patients' autoimmune manifestations, we also observed abnormal regulatory T-cell (Treg) phenotype with profoundly reduced FOXP3 and IKZF2 expression. Moreover, we found reduced T-cell proliferation and impaired STAT5B phosphorylation upon interleukin-2 stimulation of the patients' lymphocytes. In conclusion, DOCK11 deficiency is a new X-linked immune-related actinopathy leading to impaired CDC42 activity and STAT5 activation, and is associated with abnormal actin cytoskeleton remodeling as well as Treg phenotype, culminating in immune dysregulation and severe early-onset autoimmunity.
    MeSH term(s) Humans ; Male ; Actin Cytoskeleton/metabolism ; Autoimmunity ; Guanine Nucleotide Exchange Factors/genetics ; Guanine Nucleotide Exchange Factors/metabolism ; Immune System Diseases/metabolism ; Immunologic Deficiency Syndromes/complications ; Immunologic Deficiency Syndromes/genetics ; T-Lymphocytes, Regulatory
    Chemical Substances DOCK8 protein, human ; Guanine Nucleotide Exchange Factors ; DOCK11 protein, human
    Language English
    Publishing date 2023-03-23
    Publishing country United States
    Document type Case Reports ; Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 80069-7
    ISSN 1528-0020 ; 0006-4971
    ISSN (online) 1528-0020
    ISSN 0006-4971
    DOI 10.1182/blood.2022018486
    Database MEDical Literature Analysis and Retrieval System OnLINE

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