TY - JOUR A1 - Liu, Fengming A1 - Han, Kun A1 - Blair, Robert A1 - Kenst, Kornelia A1 - Qin, Zhongnan A1 - Upcin, Berin A1 - Wörsdörfer, Philipp A1 - Midkiff, Cecily C. A1 - Mudd, Joseph A1 - Belyaeva, Elizaveta A1 - Milligan, Nicholas S. A1 - Rorison, Tyler D. A1 - Wagner, Nicole A1 - Bodem, Jochen A1 - Dölken, Lars A1 - Aktas, Bertal H. A1 - Vander Heide, Richard S. A1 - Yin, Xiao-Ming A1 - Kolls, Jay K. A1 - Roy, Chad J. A1 - Rappaport, Jay A1 - Ergün, Süleyman A1 - Qin, Xuebin T1 - SARS-CoV-2 Infects Endothelial Cells In Vivo and In Vitro JF - Frontiers in Cellular and Infection Microbiology N2 - SARS-CoV-2 infection can cause fatal inflammatory lung pathology, including thrombosis and increased pulmonary vascular permeability leading to edema and hemorrhage. In addition to the lung, cytokine storm-induced inflammatory cascade also affects other organs. SARS-CoV-2 infection-related vascular inflammation is characterized by endotheliopathy in the lung and other organs. Whether SARS-CoV-2 causes endotheliopathy by directly infecting endothelial cells is not known and is the focus of the present study. We observed 1) the co-localization of SARS-CoV-2 with the endothelial cell marker CD31 in the lungs of SARS-CoV-2-infected mice expressing hACE2 in the lung by intranasal delivery of adenovirus 5-hACE2 (Ad5-hACE2 mice) and non-human primates at both the protein and RNA levels, and 2) SARS-CoV-2 proteins in endothelial cells by immunogold labeling and electron microscopic analysis. We also detected the co-localization of SARS-CoV-2 with CD31 in autopsied lung tissue obtained from patients who died from severe COVID-19. Comparative analysis of RNA sequencing data of the lungs of infected Ad5-hACE2 and Ad5-empty (control) mice revealed upregulated KRAS signaling pathway, a well-known pathway for cellular activation and dysfunction. Further, we showed that SARS-CoV-2 directly infects mature mouse aortic endothelial cells (AoECs) that were activated by performing an aortic sprouting assay prior to exposure to SARS-CoV-2. This was demonstrated by co-localization of SARS-CoV-2 and CD34 by immunostaining and detection of viral particles in electron microscopic studies. Moreover, the activated AoECs became positive for ACE-2 but not quiescent AoECs. Together, our results indicate that in addition to pneumocytes, SARS-CoV-2 also directly infects mature vascular endothelial cells in vivo and ex vivo, which may contribute to cardiovascular complications in SARS-CoV-2 infection, including multipleorgan failure. KW - endothelial cell infection KW - animal models KW - SARS-CoV-2 KW - aorta ring KW - hACE2 Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-241948 SN - 2235-2988 VL - 11 ER - TY - JOUR A1 - Dogan, Leyla A1 - Scheuring, Ruben A1 - Wagner, Nicole A1 - Ueda, Yuichiro A1 - Schmidt, Sven A1 - Wörsdörfer, Philipp A1 - Groll, Jürgen A1 - Ergün, Süleyman T1 - Human iPSC-derived mesodermal progenitor cells preserve their vasculogenesis potential after extrusion and form hierarchically organized blood vessels JF - Biofabrication N2 - Post-fabrication formation of a proper vasculature remains an unresolved challenge in bioprinting. Established strategies focus on the supply of the fabricated structure with nutrients and oxygen and either rely on the mere formation of a channel system using fugitive inks or additionally use mature endothelial cells and/or peri-endothelial cells such as smooth muscle cells for the formation of blood vessels in vitro. Functional vessels, however, exhibit a hierarchical organization and multilayered wall structure that is important for their function. Human induced pluripotent stem cell-derived mesodermal progenitor cells (hiMPCs) have been shown to possess the capacity to form blood vessels in vitro, but have so far not been assessed for their applicability in bioprinting processes. Here, we demonstrate that hiMPCs, after formulation into an alginate/collagen type I bioink and subsequent extrusion, retain their ability to give rise to the formation of complex vessels that display a hierarchical network in a process that mimics the embryonic steps of vessel formation during vasculogenesis. Histological evaluations at different time points of extrusion revealed the initial formation of spheres, followed by lumen formation and further structural maturation as evidenced by building a multilayered vessel wall and a vascular network. These findings are supported by immunostainings for endothelial and peri-endothelial cell markers as well as electron microscopic analyses at the ultrastructural level. Moreover, endothelial cells in capillary-like vessel structures deposited a basement membrane-like matrix at the basal side between the vessel wall and the alginate-collagen matrix. After transplantation of the printed constructs into the chicken chorioallantoic membrane (CAM) the printed vessels connected to the CAM blood vessels and get perfused in vivo. These results evidence the applicability and great potential of hiMPCs for the bioprinting of vascular structures mimicking the basic morphogenetic steps of de novo vessel formation during embryogenesis. KW - vascular biofabrication KW - human iPSC-derived mesodermal cells (hiMPCs) KW - extrusion of hiMPC-containing bioinks alginate + collagen type I KW - multilayered vessel wall with intimate, media and adventitia KW - vascular network and hierarchical organized vessels KW - electron microscopy KW - serial block face EM Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-254046 VL - 13 IS - 4 ER - TY - JOUR A1 - Koeniger, Tobias A1 - Bell, Luisa A1 - Mifka, Anika A1 - Enders, Michael A1 - Hautmann, Valentin A1 - Mekala, Subba Rao A1 - Kirchner, Philipp A1 - Ekici, Arif B. A1 - Schulz, Christian A1 - Wörsdörfer, Philipp A1 - Mencl, Stine A1 - Kleinschnitz, Christoph A1 - Ergün, Süleyman A1 - Kuerten, Stefanie T1 - Bone marrow‐derived myeloid progenitors in the leptomeninges of adult mice JF - Stem Cells N2 - Although the bone marrow contains most hematopoietic activity during adulthood, hematopoietic stem and progenitor cells can be recovered from various extramedullary sites. Cells with hematopoietic progenitor properties have even been reported in the adult brain under steady‐state conditions, but their nature and localization remain insufficiently defined. Here, we describe a heterogeneous population of myeloid progenitors in the leptomeninges of adult C57BL/6 mice. This cell pool included common myeloid, granulocyte/macrophage, and megakaryocyte/erythrocyte progenitors. Accordingly, it gave rise to all major myelo‐erythroid lineages in clonogenic culture assays. Brain‐associated progenitors persisted after tissue perfusion and were partially inaccessible to intravenous antibodies, suggesting their localization behind continuous blood vessel endothelium such as the blood‐arachnoid barrier. Flt3\(^{Cre}\) lineage tracing and bone marrow transplantation showed that the precursors were derived from adult hematopoietic stem cells and were most likely continuously replaced via cell trafficking. Importantly, their occurrence was tied to the immunologic state of the central nervous system (CNS) and was diminished in the context of neuroinflammation and ischemic stroke. Our findings confirm the presence of myeloid progenitors at the meningeal border of the brain and lay the foundation to unravel their possible functions in CNS surveillance and local immune cell production. KW - hematopoietic KW - meninges KW - mouse KW - myeloid KW - progenitor Y1 - 2021 U6 - http://nbn-resolving.de/urn/resolver.pl?urn:nbn:de:bvb:20-opus-224452 VL - 39 IS - 2 SP - 227 EP - 239 ER -