TY - JOUR AB - To improve outcomes following lung transplantation, it is essential to understand the immunological mechanisms that result in chronic graft failure. The associated clinical syndrome is termed chronic lung allograft dysfunction (CLAD), which is known to be induced by alloimmune-dependent (i.e., rejection) and alloimmune-independent factors (e.g., infections, reflux and environmental factors). We aimed to explore the alloimmune-related mechanism, i.e., pulmonary rejection. In this study, we use a murine orthotopic left lung transplant model using isografts and allografts (C57BL/6 or BALB/c as donors to C57BL/6 recipients), with daily immunosuppression (10 mg/kg cyclosporin A and 1.6 mg/kg methylprednisolone). Serial sacrifice was performed at days 1, 7 and 35 post-transplantation (n = 6 at each time point for each group). Left transplanted lungs were harvested, a single-cell suspension was made and absolute numbers of immune cells were quantified using multicolor flow cytometry. The rejection process followed the principles of a classic immune response, including innate but mainly adaptive immune cells. At day 7 following transplantation, the numbers of interstitial macrophages, monocytes, dendritic cells, NK cells, NKT cells, CD4+ T cells and CD8+ T and B cells were increased in allografts compared with isografts. Only dendritic cells and CD4+ T cells remained elevated at day 35 in allografts. Our study provides insights into the immunological mechanisms of true pulmonary rejection after murine lung transplantation. These results might be important in further research on diagnostic evaluation and treatment for CLAD. AU - Kaes, J.* AU - Pollenus, E.* AU - Hooft, C.* AU - Liu, H. AU - Aelbrecht, C.* AU - Cambier, S.* AU - Jin, X.* AU - Van Slambrouck, J.* AU - Beeckmans, H.* AU - Kerckhof, P.* AU - Velde, G.V.* AU - Van Raemdonck, D.* AU - Yildirim, A.Ö. AU - Van den Steen, P.E.* AU - Vos, R.* AU - Ceulemans, L.J.* AU - Vanaudenaerde, B.M.* C1 - 69945 C2 - 55331 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - The immunopathology of pulmonary rejection after murine lung transplantation. JO - Cells VL - 13 IS - 3 PB - Mdpi PY - 2024 SN - 2073-4409 ER - TY - JOUR AB - The membrane composition of extracellular vesicles (EVs) largely reflects that of the plasma membrane of the cell of origin. We therefore hypothesized that EVs could be used for immunizations to generate monoclonal antibodies against well-known tumor antigens but possibly also against hitherto unknown tumor-associated target molecules. From an immunization experiment, we obtained a monoclonal antibody specific for SRRM2, an RNA-binding protein involved in splicing and a major component of nuclear speckles. Here, we used this antibody to demonstrate that SRRM2 is exposed on the surface of most cancer cell lines from various entities and, even more important, on cancer cells in vivo. Moreover, we demonstrated that SRRM2-specific CAR-T cells are functional in vitro and in vivo. Collectively, we identified SRRM2 as a promising new target molecule exposed on the cancer cell surface and showed that our SRRM2-specific antibody can be used as a basis for the development of new targeted cancer therapies. AU - Kellner, M. AU - Hörmann, J. AU - Fackler, S. AU - Hu, Y.* AU - Zhou, T.* AU - Lu, L.* AU - Ilik, I.* AU - Aktas, T.* AU - Feederle, R. AU - Hauck, S.M. AU - Gires, O.* AU - Gärtner, K.* AU - Li, L.* AU - Zeidler, R. C1 - 71846 C2 - 56451 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - The nuclear speckles protein SRRM2 is exposed on the surface of cancer cells. JO - Cells VL - 13 IS - 18 PB - Mdpi PY - 2024 SN - 2073-4409 ER - TY - JOUR AB - Aging is the main risk factor for chronic lung diseases (CLDs) including idiopathic pulmonary fibrosis (IPF) and chronic obstructive pulmonary disease (COPD). Accordingly, hallmarks of aging like cellular senescence are increased in these patients in different lung cell types including fibroblasts. However, little is known about the different triggers that induce a senescence phenotype in different disease backgrounds and its role in CLD pathogenesis. Therefore, we characterized senescence in primary human lung fibroblasts (phLF) from control, IPF, or COPD patients at baseline and after exposure to disease-relevant insults (H2O2, bleomycin, TGF-β1) and studied their capacity to support progenitor cell potential in a lung organoid model. Bulk-RNA sequencing revealed that phLF from IPF and COPD activate different transcriptional programs but share a similar senescence phenotype at baseline. Moreover, H2O2 and bleomycin but not TGF-β1 induced senescence in phLF from different disease origins. Exposure to different triggers resulted in distinct senescence programs in phLF characterized by different SASP profiles. Finally, co-culture with bleomycin- and H2O2-treated phLF reduced the progenitor cell potential of alveolar epithelial progenitor cells. In conclusion, phLF from COPD and IPF share a conserved senescence response that varies depending on the insult and impairs alveolar epithelial progenitor capacity ex vivo. AU - Melo-Narváez, M.C AU - Bramey, N. AU - See, F. AU - Heinzelmann, K. AU - Ballester, B. AU - Steinchen, C. AU - Jain, E. AU - Federl, K. AU - Hu, Q.* AU - Dhakad, D. AU - Behr, J.* AU - Eickelberg, O.* AU - Yildirim, A.Ö. AU - Königshoff, M.* AU - Lehmann, M. C1 - 71113 C2 - 56019 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Stimuli-specific senescence of primary human lung fibroblasts modulates alveolar stem cell function. JO - Cells VL - 13 IS - 13 PB - Mdpi PY - 2024 SN - 2073-4409 ER - TY - JOUR AB - Direct neuronal reprogramming is a promising approach to replace neurons lost due to disease via the conversion of endogenous glia reacting to brain injury into neurons. However, it is essential to demonstrate that the newly generated neurons originate from glial cells and/or show that they are not pre-existing endogenous neurons. Here, we use controls for both requirements while comparing two viral vector systems (Mo-MLVs and AAVs) for the expression of the same neurogenic factor, the phosphorylation-resistant form of Neurogenin2. Our results show that Mo-MLVs targeting proliferating glial cells after traumatic brain injury reliably convert astrocytes into neurons, as assessed by genetic fate mapping of astrocytes. Conversely, expressing the same neurogenic factor in a flexed AAV system results in artefactual labelling of endogenous neurons fatemapped by birthdating in development that are negative for the genetic fate mapping marker induced in astrocytes. These results are further corroborated by chronic live in vivo imaging. Taken together, the phosphorylation-resistant form of Neurogenin2 is more efficient in reprogramming reactive glia into neurons than its wildtype counterpart in vivo using retroviral vectors (Mo-MLVs) targeting proliferating glia. Conversely, AAV-mediated expression generates artefacts and is not sufficient to achieve fate conversion. AU - Puglisi, M. AU - Lao, C.L. AU - Wani, G. AU - Masserdotti, G. AU - Bocchi, R. AU - Götz, M. C1 - 71720 C2 - 56384 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Comparing viral vectors and fate mapping approaches for astrocyte-to-neuron reprogramming in the injured mouse cerebral cortex. JO - Cells VL - 13 IS - 17 PB - Mdpi PY - 2024 SN - 2073-4409 ER - TY - JOUR AB - Antiviral neutralizing antibodies (nAbs) are commonly derived from B cells developed in immunized or infected animals and humans. Fully human antibodies are preferred for clinical use as they are potentially less immunogenic. However, the function of B cells varies depending on their homing pattern and an additional hurdle for antibody discovery in humans is the source of human tissues with an immunological microenvironment. Here, we show an efficient method to pharm human antibodies using immortalized B cells recovered from Nod.Rag.Gamma (NRG) mice reconstituting the human immune system (HIS). Humanized HIS mice were immunized either with autologous engineered dendritic cells expressing the human cytomegalovirus gB envelope protein (HCMV-gB) or with Epstein–Barr virus-like particles (EB-VLP). Human B cells recovered from spleen of HIS mice were efficiently immortalized with EBV in vitro. We show that these immortalized B cells secreted human IgGs with neutralization capacities against prototypic HCMV-gB and EBV-gp350. Taken together, we show that HIS mice can be successfully used for the generation and pharming fully human IgGs. This technology can be further explored to generate antibodies against emerging infections for diagnostic or therapeutic purposes. AU - Theobald, S.J.* AU - Fiestas Carcaba, E. AU - Schneider, A.* AU - Ostermann, B.* AU - Danisch, S.* AU - von Kaisenberg, C.* AU - Rybniker, J.* AU - Hammerschmidt, W. AU - Zeidler, R. AU - Stripecke, R.* C1 - 69730 C2 - 53863 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Fully human herpesvirus-specific neutralizing IgG antibodies generated by EBV immortalization of splenocytes-derived from immunized humanized Mice. JO - Cells VL - 13 IS - 1 PB - Mdpi PY - 2024 SN - 2073-4409 ER - TY - JOUR AB - Conventional 2D cultures are commonly used in cancer research though they come with limitations such as the lack of microenvironment or reduced cell heterogeneity. In this study, we investigated in what respect a scaffold-based (Matrigel™) 3D culture technique can ameliorate the limitations of 2D cultures. NGS-based bulk and single-cell sequencing of matched pairs of 2D and 3D models showed an altered transcription of key immune regulatory genes in around 36% of 3D models, indicating the reoccurrence of an immune suppressive phenotype. Changes included the presentation of different HLA surface molecules as well as cellular stressors. We also investigated the 3D tumor organoids in a co-culture setting with tumor-infiltrating lymphocytes (TILs). Of note, lymphocyte-mediated cell killing appeared less effective in clearing 3D models than their 2D counterparts. IFN-γ release, as well as live cell staining and proliferation analysis, pointed toward an elevated resistance of 3D models. In conclusion, we found that the scaffold-based (Matrigel™) 3D culture technique affects the transcriptional profile in a subset of GBM models. Thus, these models allow for depicting clinically relevant aspects of tumor-immune interaction, with the potential to explore immunotherapeutic approaches in an easily accessible in vitro system. AU - Braun, F.K.* AU - Rothhammer-Hampl, T.* AU - Lorenz, J.* AU - Pohl, S.* AU - Menevse, A.N.* AU - Vollmann-Zwerenz, A.* AU - Bumes, E.* AU - Büttner, M. AU - Zoubaa, S.* AU - Proescholdt, M.* AU - Schmidt, N.O.* AU - Hau, P.* AU - Beckhove, P.* AU - Winner, B.* AU - Riemenschneider, M.J.* C1 - 68055 C2 - 54533 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Scaffold-based (Matrigel™) 3D culture technique of glioblastoma recovers a patient-like immunosuppressive phenotype. JO - Cells VL - 12 IS - 14 PB - Mdpi PY - 2023 SN - 2073-4409 ER - TY - JOUR AB - The cultivation of marine invertebrate cells in vitro has garnered significant attention due to the availability of diverse cell types and cellular potentialities in comparison to vertebrates and particularly in response to the demand for a multitude of applications. While cells in the colonial urochordate Botryllus schlosseri have a very high potential for omnipotent differentiation, no proliferating cell line has been established in Botryllus, with results indicating that cell divisions cease 24-72 h post initiation. This research assessed how various Botryllus blood cell types respond to in vitro conditions by utilizing five different refinements of cell culture media (TGM1-TGM5). During the initial week of culture, there was a noticeable medium-dependent increase in the proliferation and viability of distinct blood cell types. Within less than one month from initiation, we developed medium-specific primary cultures, a discovery that supports larger efforts to develop cell type-specific cultures. Specific cell types were easily distinguished and classified based on their natural fluorescence properties using confocal microscopy. These results are in agreement with recent advances in marine invertebrate cell cultures, demonstrating the significance of optimized nutrient media for cell culture development and for cell selection. AU - Qarri, A. AU - Kültz, D.* AU - Gardell, A.M.* AU - Rinkevich, B.* AU - Rinkevich, Y. C1 - 68107 C2 - 54585 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Improved media formulations for primary cell cultures derived from a colonial urochordate. JO - Cells VL - 12 IS - 13 PB - Mdpi PY - 2023 SN - 2073-4409 ER - TY - JOUR AB - CD30-positive germinal center (GC)-derived B cell lymphomas are frequently linked to Epstein-Barr Virus (EBV) infection. However, a suitable animal model for the investigation of the interplay between γ-herpesvirus and host cells in B cell pathogenesis is currently lacking. Here, we present a novel in vivo model enabling the analysis of genetically modified viruses in combination with genetically modified GC B cells. As a murine γ-herpesvirus, we used MHV-68 closely mirroring the biology of EBV. Our key finding was that Cre-mediated recombination can be successfully induced by an MHV-68 infection in GC B cells from Cγ1-Cre mice allowing for deletion or activation of loxP-flanked cellular genes. The implementation of PrimeFlow RNA assay for MHV-68 demonstrated the enrichment of MHV-68 in GC and isotype-switched B cells. As illustrations of virus and cellular modifications, we inserted the EBV gene LMP2A into the MHV-68 genome and induced constitutively active CD30-signaling in GC B cells through MHV-68 infections, respectively. While the LMP2A-expressing MHV-68 behaved similarly to wildtype MHV-68, virally induced constitutively active CD30-signaling in GC B cells led to the expansion of a pre-plasmablastic population. The findings underscore the potential of our novel tools to address crucial questions about the interaction between herpesviral infections and deregulated cellular gene-expression in future studies. AU - Rambold, U. AU - Sperling, S. AU - Chew, Z. AU - Wang, Y. AU - Steer, B. AU - Zeller, K. AU - Strobl, L.J. AU - Zimber-Strobl, U. AU - Adler, H. C1 - 69045 C2 - 53827 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - A mouse model to study the pathogenesis of γ-herpesviral infections in germinal center B cells. JO - Cells VL - 12 IS - 24 PB - Mdpi PY - 2023 SN - 2073-4409 ER - TY - JOUR AB - The human dopaminergic system is vital for a broad range of neurological processes, including the control of voluntary movement. Here we report a proband presenting with clinical features of dopamine deficiency: severe infantile parkinsonism-dystonia, characterised by frequent oculogyric crises, dysautonomia and global neurodevelopmental impairment. CSF neurotransmitter analysis was unexpectedly normal. Triome whole-genome sequencing revealed a homozygous variant (c.110C>A, (p.T37K)) in DRD1, encoding the most abundant dopamine receptor (D1) in the central nervous system, most highly expressed in the striatum. This variant was absent from gnomAD, with a CADD score of 27.5. Using an in vitro heterologous expression system, we determined that DRD1-T37K results in loss of protein function. Structure-function modelling studies predicted reduced substrate binding, which was confirmed in vitro. Exposure of mutant protein to the selective D1 agonist Chloro APB resulted in significantly reduced cyclic AMP levels. Numerous D1 agonists failed to rescue the cellular defect, reflected clinically in the patient, who had no benefit from dopaminergic therapy. Our study identifies DRD1 as a new disease-associated gene, suggesting a crucial role for the D1 receptor in motor control. AU - Reid, K.M.* AU - Steel, D.* AU - Nair, S.* AU - Bhate, S.* AU - Biassoni, L.* AU - Sudhakar, S.* AU - Heys, M.* AU - Burke, E.* AU - Kamsteeg, E.J.* AU - Hameed, B.* AU - Zech, M. AU - Mencacci, N.E.* AU - Barwick, K.* AU - Topf, M.* AU - Kurian, M.A.* C1 - 67612 C2 - 53919 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Loss-of-function variants in DRD1 in infantile parkinsonism-dystonia. JO - Cells VL - 12 IS - 7 PB - Mdpi PY - 2023 SN - 2073-4409 ER - TY - JOUR AB - Amyloid-β (Aβ) deposition is an initiating factor in Alzheimer's disease (AD). Microglia are the brain immune cells that surround and phagocytose Aβ plaques, but their phagocytic capacity declines in AD. This is in agreement with studies that associate AD risk loci with genes regulating the phagocytic function of immune cells. Immunotherapies are currently pursued as strategies against AD and there are increased efforts to understand the role of the immune system in ameliorating AD pathology. Here, we evaluated the effect of the Aβ targeting ACI-24 vaccine in reducing AD pathology in an amyloidosis mouse model. ACI-24 vaccination elicited a robust and sustained antibody response in APPPS1 mice with an accompanying reduction of Aβ plaque load, Aβ plaque-associated ApoE and dystrophic neurites as compared to non-vaccinated controls. Furthermore, an increased number of NLRP3-positive plaque-associated microglia was observed following ACI-24 vaccination. In contrast to this local microglial activation at Aβ plaques, we observed a more ramified morphology of Aβ plaque-distant microglia compared to non-vaccinated controls. Accordingly, bulk transcriptomic analysis revealed a trend towards the reduced expression of several disease-associated microglia (DAM) signatures that is in line with the reduced Aβ plaque load triggered by ACI-24 vaccination. Our study demonstrates that administration of the Aβ targeting vaccine ACI-24 reduces AD pathology, suggesting its use as a safe and cost-effective AD therapeutic intervention. AU - Rudan Njavro, J.* AU - Vukicevic, M.* AU - Fiorini, E.* AU - Dinkel, L.* AU - Müller, S.A.* AU - Berghofer, A.* AU - Bordier, C.* AU - Kozlov, S.* AU - Halle, A.* AU - Buschmann, K.* AU - Capell, A.* AU - Giudici, C.* AU - Willem, M.* AU - Feederle, R. AU - Lichtenthaler, S.F.* AU - Babolin, C.* AU - Montanari, P.* AU - Pfeifer, A.* AU - Kosco-Vilbois, M.* AU - Tahirovic, S.* C1 - 67178 C2 - 53517 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Beneficial effect of ACI-24 vaccination on Aβ plaque pathology and microglial phenotypes in an amyloidosis mouse model. JO - Cells VL - 12 IS - 1 PB - Mdpi PY - 2023 SN - 2073-4409 ER - TY - JOUR AB - The authors made the following changes to their paper [1]. The authors noticed a mistake on the SF-1 results for the cell line NCI-H295 in Figure 6. A mistake happened during data transfer for graph preparation. The authors are very sorry about this error. They have now repeated the specific experiment for a total of five times; it was performed by two independent persons and with two different primer pairs (only the original primer pair is included here, but two different primer pairs were used to ensure that the effect between different SF-1 primers is consistent). Figure 6I (marked in red) was corrected from to A correction has been made to Section 3.7: The sentence “In contrast, HSD17B4 remained unchanged and the AR, ER1 and SF-1 were conversely down-regulated for NCI-H295R.For TVBF-7, again, all levels remained unchanged” was modified to “In contrast, HSD17B4 remained unchanged, SF-1 was upregulated and AR and ER1 were conversely down-regulated for NCI-H295R. For TVBF-7, again, all levels remained unchanged”. A correction has been made to Section 4, paragraphs 4 and 5: The sentence “Of note, SF-1 is a key regulator of human sex determination [39] and was upon FSK stimulation strikingly different regulated in NCI-H295R (down), MUC-1 (up) and TVBF-7 (unchanged)” was corrected to “Of note, SF-1 is a key regulator of human sex determination [39] and its activation might lead to different downstream effects in tissues of male and female origin”. The sentence “Of note, the AR was again markedly different regulated following the same patterns as observed for SF-1 in NCI-H295R (down), MUC-1 (up) and TVBF-7 (unchanged)” was corrected to “Of note, the AR was markedly differently regulated in NCI-H295R (down), MUC-1 (up) and TVBF-7 (unchanged)”. The authors state that the scientific conclusions are unaffected. This correction was approved by the Academic Editor. The original publication has also been updated. AU - Sigala, S.* AU - Bothou, C.* AU - Penton, D.* AU - Abate, A.* AU - Peitzsch, M.* AU - Cosentini, D.* AU - Tiberio, G.A.M.* AU - Bornstein, S.R. AU - Berruti, A.* AU - Hantel, C.* C1 - 68211 C2 - 54830 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Correction to: A Comprehensive Investigation of Steroidogenic Signaling in Classical and New Experimental Cell Models of Adrenocortical Carcinoma (Cells, (2022), 11, 9, (1439), 10.3390/cells11091439). JO - Cells VL - 12 IS - 18 PB - Mdpi PY - 2023 SN - 2073-4409 ER - TY - JOUR AB - Primary ciliary dyskinesia (PCD) is a rare heterogenic genetic disorder associated with perturbed biogenesis or function of motile cilia. Motile cilia dysfunction results in diminished mucociliary clearance (MCC) of pathogens in the respiratory tract and chronic airway inflammation and infections successively causing progressive lung damage. Current approaches to treat PCD are symptomatic, only, indicating an urgent need for curative therapeutic options. Here, we developed an in vitro model for PCD based on human induced pluripotent stem cell (hiPSC)-derived airway epithelium in Air-Liquid-Interface cultures. Applying transmission electron microscopy, immunofluorescence staining, ciliary beat frequency, and mucociliary transport measurements, we could demonstrate that ciliated respiratory epithelia cells derived from two PCD patient-specific hiPSC lines carrying mutations in DNAH5 and NME5, respectively, recapitulate the respective diseased phenotype on a molecular, structural and functional level. AU - von Schledorn, L.* AU - Puertollano Martín, D.* AU - Cleve, N.* AU - Zöllner, J.* AU - Roth, D. AU - Staar, B.O.* AU - Hegermann, J.* AU - Ringshausen, F.C.* AU - Nawroth, J. AU - Martin, U.* AU - Olmer, R.* C1 - 67834 C2 - 54312 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Primary ciliary dyskinesia patient-specific hiPSC-derived airway epithelium in air-liquid interface culture recapitulates disease specific phenotypes in vitro. JO - Cells VL - 12 IS - 11 PB - Mdpi PY - 2023 SN - 2073-4409 ER - TY - JOUR AB - Since their initial description by Elie Metchnikoff, phagocytes have sparked interest in a variety of biologic disciplines. These important cells perform central functions in tissue repair and immune activation as well as tolerance. Myeloid cells can be immunoinhibitory, particularly in the tumor microenvironment, where their presence is generally associated with poor patient prognosis. These cells are highly adaptable and plastic, and can be modulated to perform desired functions such as antitumor activity, if key programming molecules can be identified. Human clear cell renal cell carcinoma (ccRCC) is considered immunogenic; yet checkpoint blockades that target T cell dysfunction have shown limited clinical efficacy, suggesting additional layers of immunoinhibition. We previously described “enriched-in-renal cell carcinoma” (erc) DCs that were often found in tight contact with dysfunctional T cells. Using transcriptional profiling and flow cytometry, we describe here that ercDCs represent a mosaic cell type within the macrophage continuum co-expressing M1 and M2 markers. The polarization state reflects tissue-specific signals that are characteristic of RCC and renal tissue homeostasis. ErcDCs are tissue-resident with increasing prevalence related to tumor grade. Accordingly, a high ercDC score predicted poor patient survival. Within the profile, therapeutic targets (VSIG4, NRP1, GPNMB) were identified with promise to improve immunotherapy. AU - Brech, D. AU - Herbstritt, A. AU - Diederich, S. AU - Straub, T.* AU - Kokolakis, E. AU - Irmler, M. AU - Beckers, J. AU - Büttner, F.A.* AU - Schaeffeler, E.* AU - Winter, S.* AU - Nelson, P.J.* AU - Nößner, E. C1 - 66541 C2 - 52891 TI - Dendritic cells or macrophages? The microenvironment of human clear cell renal cell carcinoma imprints a mosaic myeloid subtype associated with patient survival. JO - Cells VL - 11 IS - 20 PY - 2022 SN - 2073-4409 ER - TY - JOUR AB - Low-grade chronic inflammation and reduced differentiation capacity are hallmarks of hy-pertrophic adipose tissue (AT) and key contributors of insulin resistance. We identified PPARG∆5 as a dominant-negative splicing isoform overexpressed in the AT of obese/diabetic patients able to impair adipocyte differentiation and PPARγ activity in hypertrophic adipocytes. Herein, we investigate the impact of macrophage-secreted pro-inflammatory factors on PPARG splicing, focusing on PPARG∆5. We report that the epididymal AT of LPS-treated mice displays increased Pparg∆5/cPparg ratio and reduced expression of Pparg-regulated genes. Interestingly, pro-inflammatory factors secreted from murine and human pro-inflammatory macrophages enhance the PPARG∆5/cPPARG ratio in exposed adipogenic precursors. TNFα is identified herein as factor able to alter PPARG splicing— increasing PPARG∆5/cPPARG ratio—through PI3K/Akt signaling and SRp40 splicing factor. In line with in vitro data, TNFA expression is higher in the SAT of obese (vs. lean) patients and posi-tively correlates with PPARG∆5 levels. In conclusion, our results indicate that inflammatory factors secreted by metabolically-activated macrophages are potent stimuli that modulate the expression and splicing of PPARG. The resulting imbalance between canonical and dominant negative isoforms may crucially contribute to impair PPARγ activity in hypertrophic AT, exacerbating the defective adipogenic capacity of precursor cells. AU - Cataldi, S.* AU - Aprile, M.* AU - Melillo, D.* AU - Mucel, I.* AU - Giorgetti-Peraldi, S.* AU - Cormont, M.* AU - Italiani, P.* AU - Blüher, M. AU - Tanti, J.F.* AU - Ciccodicola, A.* AU - Costa, V.* C1 - 63926 C2 - 51657 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - TNFα mediates inflammation-induced effects on PPARG splicing in adipose tissue and mesenchymal precursor cells. JO - Cells VL - 11 IS - 1 PB - Mdpi PY - 2022 SN - 2073-4409 ER - TY - JOUR AB - Idiopathic pulmonary fibrosis (IPF) is a fatal disease with incompletely understood aetiol-ogy and limited treatment options. Traditionally, IPF was believed to be mainly caused by repetitive injuries to the alveolar epithelium. Several recent lines of evidence, however, suggest that IPF equally involves an aberrant airway epithelial response, which contributes significantly to disease development and progression. In this review, based on recent clinical, high-resolution imaging, genetic, and single-cell RNA sequencing data, we summarize alterations in airway structure, function, and cell type composition in IPF. We furthermore give a comprehensive overview on the genetic and mecha-nistic evidence pointing towards an essential role of airway epithelial cells in IPF pathogenesis and describe potentially implicated aberrant epithelial signalling pathways and regulation mechanisms in this context. The collected evidence argues for the investigation of possible therapeutic avenues targeting these processes, which thus represent important future directions of research. AU - Chakraborty, A. AU - Mastalerz, M. AU - Ansari, M. AU - Schiller, H. B. AU - Staab-Weijnitz, C.A. C1 - 64660 C2 - 52398 TI - Emerging roles of airway epithelial cells in idiopathic pulmonary fibrosis. JO - Cells VL - 11 IS - 6 PY - 2022 SN - 2073-4409 ER - TY - JOUR AB - Glioblastoma stem-like cells (GSLCs) in glioblastoma limit effective treatment and promote therapeutic resistance and tumor recurrence. Using a combined radiation and drugscreening platform, we tested the combination of a histone deacetylase inhibitor (HDACi) and MAPK/ERK kinase inhibitor (MEKi) with radiation to predict the efficacy against GSLCs. To mimic a stem-like phenotype, glioblastoma-derived spheres were used and treated with a combination of HDACi (MS-275) and MEKi (TAK-733 or trametinib) with 4 Gy irradiation. The sphere-forming ability after the combined radiochemotherapy was investigated using a sphere formation assay, while the expression levels of the GSLC markers (CD44, Nestin and SOX2) after treatment were analyzed using Western blotting and flow cytometry. The combined radiochemotherapy treatment inhibited the sphere formation in both glioblastoma-derived spheres, decreased the expression of the GSLC markers in a cell-line dependent manner and increased the dead cell population. Finally, we showed that the combined treatment with radiation was more effective at reducing the GSLC markers compared to the standard treatment of temozolomide and radiation. These results suggest that combining HDAC and MEK inhibition with radiation may offer a new strategy to improve the treatment of glioblastoma. AU - Essien, E.I. AU - Hofer, T.P. AU - Atkinson, M.J.* AU - Anastasov, N. C1 - 64464 C2 - 52225 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Combining HDAC and MEK inhibitors with radiation against glioblastoma-derived spheres. JO - Cells VL - 11 IS - 5 PB - Mdpi PY - 2022 SN - 2073-4409 ER - TY - JOUR AB - Glucocorticoids are potent anti-inflammatory drugs; however, their molecular mode of action remains complex and elusive. They bind to the glucocorticoid receptor (GR), a nuclear receptor that controls gene expression in almost all tissues in a cell type-specific manner. While GR’s transcriptional targets mediate beneficial reactions in immune cells, they also harbor the potential of adverse metabolic effects in other cell types such as hepatocytes. Here, we have profiled nascent transcription upon glucocorticoid stimulation in LPS-activated primary murine macrophages using 4sU-seq. We compared our results to publicly available nascent transcriptomics data from murine liver and bioinformatically identified non-coding RNAs transcribed from intergenic GR binding sites in a tissue-specific fashion. These tissue-specific enhancer RNAs (eRNAs) correlate with target gene expression, reflecting cell type-specific glucocorticoid responses. We further associate GRmediated eRNA expression with changes in H3K27 acetylation and BRD4 recruitment in inflammatory macrophages upon glucocorticoid treatment. In summary, we propose a common mechanism by which GR-bound enhancers regulate target gene expression by changes in histone acetylation, BRD4 recruitment and eRNA expression. We argue that local eRNAs are potential therapeutic targets downstream of GR signaling which may modulate glucocorticoid response in a cell type-specific way. AU - Greulich, F. AU - Bielefeld, K.A. AU - Scheundel, R.* AU - Mechtidou, A. AU - Strickland, B.* AU - Uhlenhaut, N.H. C1 - 63913 C2 - 51669 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Enhancer RNA expression in response to glucocorticoid treatment in murine macrophages. JO - Cells VL - 11 IS - 1 PB - Mdpi PY - 2022 SN - 2073-4409 ER - TY - JOUR AB - Continuous activation of hypoxia pathways in pheochromocytomas and paragangliomas (PPGLs) is associated with higher disease aggressiveness, for which effective treatment strategies are still missing. Most of the commonly used in vitro models lack characteristics of these pseudohypoxic tumors, including elevated expression of hypoxia-inducible factor (HIF) 2α. To address this shortcoming, we investigated whether recurrent hypoxia cycles lead to continuous activation of hypoxia pathways under normoxic conditions and whether this pseudohypoxia is associated with increased cellular aggressiveness. Rat pheochromocytoma cells (PC12) were incubated under hypoxia for 24 h every 3–4 days, up to 20 hypoxia–reoxygenation cycles, resulting in PC12 Z20 cells. PC12 Z20 control cells were obtained by synchronous cultivation under normoxia. RNA sequencing revealed upregulation of HIF2α in PC12 Z20 cells and a pseudohypoxic gene signature that overlapped with the gene signature of pseudohypoxic PPGLs. PC12 Z20 cells showed a higher growth rate, and the migration and adhesion capacity were significantly increased compared with control cells. Changes in global methylation, together with the pseudohypoxic conditions, may be responsible for the increased aggressiveness of this new model. The established sub-cell line with characteristics of pseudohypoxic PPGLs represent a complementary model for further investigations, for example, with regard to new therapeutic approaches. AU - Helm, J.* AU - Drukewitz, S.* AU - Poser, I.* AU - Richter, S.* AU - Friedemann, M.* AU - William, D.* AU - Mohr, H. AU - Nölting, S.* AU - Robledo, M.* AU - Bornstein, S.R.* AU - Eisenhofer, G.* AU - Bechmann, N.* C1 - 64280 C2 - 52170 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Treatment of pheochromocytoma cells with recurrent cycles of hypoxia: A new pseudohypoxic in vitro model. JO - Cells VL - 11 IS - 3 PB - Mdpi PY - 2022 SN - 2073-4409 ER - TY - JOUR AB - The airway epithelium provides the first line of defense to the surrounding environment. However, dysfunctions of this physical barrier are frequently observed in allergic diseases, which are tightly connected with pro-or anti-inflammatory processes. When the epithelial cells are confronted with allergens or pathogens, specific response mechanisms are set in motion, which in homeostasis, lead to the elimination of the invaders and leave permanent traces on the respiratory epithelium. However, allergens can also cause damage in the sensitized organism, which can be ascribed to the excessive immune reactions. The tight interaction of epithelial cells of the upper and lower airways with local and systemic immune cells can leave an imprint that may mirror the pathophysiology. The interaction with effector T cells, along with the macrophages, play an important role in this response, as reflected in the gene expression profiles (transcriptomes) of the epithelial cells, as well as in the secretory pattern (secretomes). Further, the storage of information from past exposures as memories within discrete cell types may allow a tissue to inform and fundamentally alter its future responses. Recently, several lines of evidence have highlighted the contributions from myeloid cells, lymphoid cells, stromal cells, mast cells, and epithelial cells to the emerging concepts of inflammatory memory and trained immunity. AU - Jakwerth, C.A. AU - Ordovas-Montanes, J.* AU - Blank, S. AU - Schmidt-Weber, C.B. AU - Zissler, U.M. C1 - 64916 C2 - 52530 TI - Role of respiratory epithelial cells in allergic diseases. JO - Cells VL - 11 IS - 9 PY - 2022 SN - 2073-4409 ER - TY - JOUR AB - Peritubular cells of the human testis form a small compartment surrounding the seminiferous tubules. They are crucial for sperm transport, and they emerge as contributors to the spermatogonial stem cell niche. They are among the least known cell types of the human body. We employed single-cell RNA sequencing of cultured human testicular peritubular cells (HTPCs), which had been isolated from testicular samples of donors with normal spermatogenesis. The significant overlap between our results and recently published ex vivo data indicates that HTPCs are a highly adequate cellular model to define and study these cells. Thus, based on the expression of several markers, HTPCs can be classified as testicular smooth muscle cells. Small differences between the in vivo/in vitro expressed genes may be due to cellular plasticity. Plasticity was also shown upon addition of FCS to the culture medium. Based on transcriptome similarities, four cellular states were identified. Further analyses confirmed the presence of known stem cell niche-relevant factors (e.g., GDNF) and identified unknown functions, e.g., the ability to produce retinoic acid. Therefore, HTPCs allow us to define the signature(s) and delineate the functions of human testicular peritubular cells. The data may also serve as a resource for future studies to better understand male (in)fertility. AU - Liebich, A.* AU - Schmid, N.* AU - Koupourtidou, C. AU - Herrmann, C.* AU - Dietrich, K.G.* AU - Welter, H.F.* AU - Ninkovic, J. AU - Mayerhofer, A.* C1 - 66769 C2 - 53300 TI - The molecular signature of human testicular peritubular cells revealed by single-cell analysis. JO - Cells VL - 12 IS - 22 PY - 2022 SN - 2073-4409 ER - TY - JOUR AB - Objective: Up-regulated expression of transcription-factor E2F1 in human visceral adipose tissue (VAT) characterizes a dysmetabolic obesity sub-phenotype. An E2F1-miRNA network has been described in multiple cancers. Here we investigated whether elevated VAT-E2F1 in obesity is associated with VAT-miRNA alterations similar to, or distinct from, those described in cancer. Furthermore, we assessed if E2F1-associated miRNA changes may contribute to the link between high- VAT-E2F1 and a dysmetabolic obesity phenotype. Methods: We assembled a cohort of patients with obesity and high-VAT-E2F1, matched by age, sex, ±BMI to patients with low-VAT-E2F1, with and without obesity (8 patients/groupX3 groups). We performed Nanostring©-based miRNA profiling of VAT samples from all 24 patients. Candidate E2F1-related miRNAs were validated by qPCR in an independent cohort of patients with extreme obesity, with or without type-2-diabetes (T2DM) (n = 20). Bioinformatic tools and manipulation of E2F1 expression in cells were used to establish the plausibility of the functional VAT-E2F1-miRNA network in obesity. Results: Among n = 798 identified miRNAs, 17 were differentially expressed in relation to E2F1 and not to obesity itself. No evidence for the cancer-related E2F1-miRNA network was identified in human VAT in obesity. In HEK293-cells, overexpression/downregulation of E2F1 correspondingly altered the expression of miRNA-206 and miRNA-210-5p, two miRNAs with reported metabolic functions consistent with those of E2F1. In VAT from both cohorts, the expression of both miRNA-206 and 210-5p intercorrelated, and correlated with the expression of E2F1. In cohort 1 we did not detect significant associations with biochemical parameters. In cohort 2 of patients with extreme obesity, all those with high VAT-E2F1 showed a diabetes-complicated obesity phenotype and higher expression of miRNA-206 and miRNA-210-5p, which also correlated with fasting glucose levels (both miRNAs) and fasting insulin (miRNA-210-5p). Conclusions: Whilst the previously described cancer-related E2F1-miRNA network does not appear to operate in VAT in obesity, miRNAs-206 and 210-5p may link high-E2F1 expression in VAT with diabetes-complicated extreme obesity phenotype. AU - Maixner, N.* AU - Haim, Y.* AU - Blüher, M. AU - Chalifa-Caspi, V.* AU - Veksler-Lublinsky, I.* AU - Makarenkov, N.* AU - Yoel, U.* AU - Bashan, N.* AU - Liberty, I.F.* AU - Kukeev, I.* AU - Dukhno, O.* AU - Levy, D.* AU - Rudich, A.* C1 - 66443 C2 - 53181 TI - Visceral adipose tissue E2F1-miRNA206/210 pathway associates with type 2 diabetes in humans with extreme obesity. JO - Cells VL - 11 IS - 19 PY - 2022 SN - 2073-4409 ER - TY - JOUR AB - Antibodies are central effectors of the adaptive immune response, widespread used therapeutics, but also potentially disease-causing biomolecules. Antibody folding catalysts in the plasma cell are incompletely defined. Idiopathic pulmonary fibrosis (IPF) is a fatal chronic lung disease with increasingly recognized autoimmune features. We found elevated expression of FK506-binding protein 11 (FKBP11) in IPF lungs where FKBP11 specifically localized to antibodyproducing plasma cells. Suggesting a general role in plasma cells, plasma cell-specific FKBP11 expression was equally observed in lymphatic tissues, and in vitro B cell to plasma cell differentiation was accompanied by induction of FKBP11 expression. Recombinant human FKBP11 was able to refold IgG antibody in vitro and inhibited by FK506, strongly supporting a function as antibody peptidyl-prolyl cis-trans isomerase. Induction of ER stress in cell lines demonstrated induction of FKBP11 in the context of the unfolded protein response in an X-box-binding protein 1 (XBP1)-dependent manner. While deficiency of FKBP11 increased susceptibility to ER stressmediated cell death in an alveolar epithelial cell line, FKBP11 knockdown in an antibody-producing hybridoma cell line neither induced cell death nor decreased expression or secretion of IgG antibody. Similarly, antibody secretion by the same hybridoma cell line was not affected by knockdown of the established antibody peptidyl-prolyl isomerase cyclophilin B. The results are consistent with FKBP11 as a novel XBP1-regulated antibody peptidyl-prolyl cis-trans isomerase and indicate significant redundancy in the ER-resident folding machinery of antibody-producing hybridoma cells. AU - Preisendörfer, S. AU - Ishikawa, Y.* AU - Hennen, E. AU - Winklmeier, S.* AU - Schupp, J.C.* AU - Knüppel, L. AU - Fernandez, I.E. AU - Binzenhöfer, L. AU - Flatley, A. AU - Juan-Guardela, B.M.* AU - Ruppert, C.* AU - Guenther, A.* AU - Frankenberger, M. AU - Hatz, R.A.* AU - Kneidinger, N.* AU - Behr, J.* AU - Feederle, R. AU - Schepers, A. AU - Hilgendorff, A. AU - Kaminski, N.* AU - Meinl, E.* AU - Bächinger, H.P.* AU - Eickelberg, O. AU - Staab-Weijnitz, C.A. C1 - 64835 C2 - 52514 TI - FK506-binding protein 11 is a novel plasma cell-specific antibody folding catalyst with increased expression in idiopathic pulmonary fibrosis. JO - Cells VL - 11 IS - 8 PY - 2022 SN - 2073-4409 ER - TY - JOUR AB - In acute lymphoblastic leukemia (ALL), conventional cell lines do not recapitulate the clonal diversity and microenvironment. Orthotopic patient-derived xenograft models (PDX) overcome these limitations and mimic the clinical situation, but molecular stability and engraftment patterns have not yet been thoroughly assessed. We herein describe and characterize the PDX generation in NSG mice. In vivo tumor cell proliferation, engraftment and location were monitored by flow cytometry and bioluminescence imaging. Leukemic cells were retransplanted for up to four passages, and comparative analyses of engraftment pattern, cellular morphology and genomic hotspot mutations were conducted. Ninety-four percent of all samples were successfully engrafted, and the xenograft velocity was dependent on the molecular subtype, outcome of the patient and transplantation passage. While BCR::ABL1 blasts were located in the spleen, KMT2A-positive cases had higher frequencies in the bone marrow. Molecular changes appeared in most model systems, with low allele frequency variants lost during primary engraftment. After the initial xenografting, however, the PDX models demonstrated high molecular stability. This protocol for reliable ALL engraftment demonstrates variability in the location and molecular signatures during serial transplantation. Thorough characterization of experimentally used PDX systems is indispensable for the correct analysis and valid data interpretation of preclinical PDX studies. AU - Richter, A.* AU - Roolf, C.* AU - Sekora, A.* AU - Knuebel, G.* AU - Krohn, S.* AU - Lange, S.* AU - Krebs, V.* AU - Schneider, B.* AU - Lakner, J.* AU - Wittke, C.* AU - Kiefel, C.* AU - Jeremias, I. AU - Escobar, H.M.* AU - Vollmar, B.* AU - Junghanss, C.* C1 - 63950 C2 - 52035 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - The molecular subtype of adult acute lymphoblastic leukemia samples determines the engraftment site and proliferation kinetics in patient-derived xenograft models. JO - Cells VL - 11 IS - 1 PB - Mdpi PY - 2022 SN - 2073-4409 ER - TY - JOUR AB - The oligodendrocyte progenitors (OPCs) are at the front of the glial reaction to the traumatic brain injury. However, regulatory pathways steering the OPC reaction as well as the role of reactive OPCs remain largely unknown. Here, we compared a long-lasting, exacerbated reaction of OPCs to the adult zebrafish brain injury with a timely restricted OPC activation to identify the specific molecular mechanisms regulating OPC reactivity and their contribution to regeneration. We demonstrated that the influx of the cerebrospinal fluid into the brain parenchyma after injury simultaneously activates the toll-like receptor 2 (Tlr2) and the chemokine receptor 3 (Cxcr3) innate immunity pathways, leading to increased OPC proliferation and thereby exacerbated glial reactivity. These pathways were critical for long-lasting OPC accumulation even after the ablation of microglia and infiltrating monocytes. Importantly, interference with the Tlr1/2 and Cxcr3 pathways after injury alleviated reactive gliosis, increased new neuron recruitment, and improved tissue restoration. AU - Sanchez-Gonzalez, R. AU - Koupourtidou, C. AU - Lepko, T. AU - Zambusi, A. AU - Novoselc, K.T. AU - Durovic, T. AU - Aschenbroich, S. AU - Schwarz, V. AU - Breunig, C. AU - Straka, H.* AU - Huttner, H.B.* AU - Irmler, M. AU - Beckers, J. AU - Wurst, W. AU - Zwergal, A.* AU - Schauer, T.* AU - Straub, T.* AU - Czopka, T.* AU - Trümbach, D. AU - Götz, M. AU - Stricker, S.H. AU - Ninkovic, J. C1 - 64216 C2 - 52023 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Innate immune pathways promote oligodendrocyte progenitor cell recruitment to the injury site in adult Zebrafish brain. JO - Cells VL - 11 IS - 3 PB - Mdpi PY - 2022 SN - 2073-4409 ER - TY - JOUR AB - Adrenocortical carcinoma is a heterogeneous and aggressive cancer that originates from steroidogenic cells within the adrenal cortex. In this study, we have assessed for the preclinical gold standard NCI-H295 in direct comparison with the more recently established MUC-1 and a here newly reported ACC cell line (TVBF-7) the mutational status of important driver genes (TP53, MEN1, PRKAR1A, CTNNB1, APC, ZNRF-3, IGF-2, EGFR, RB1, BRCA1, BRCA2, RET, GNAS and PTEN), Wnt-signaling specificities (CTNNB1 mutation vs. APC mutation vs. wildtype), steroidogenic-(CYP11A1, CYP17A1, HSD3B2, HSD17B4, CYP21A2, CYP11B1, CYP11B2, MC2R, AT1R) and nuclear-receptor-signaling (AR, ER, GCR), varying electrophysiological potentials as well as highly individual hormone secretion profiles (Cortisol, Aldosterone, DHEA, DHEAS, Testosterone, 17-OH Progesterone, among others) which were investigated under basal and stimulated conditions (ACTH, AngII, FSK). Our findings reveal important genetic and pathophysiological characteristics for these three cell lines and reveal the importance of such cell-line panels reflecting differential endocrine functionalities to thereby better reflect clinically well-known ACC patient heterogeneities in preclinical studies. AU - Sigala, S.* AU - Bothou, C.* AU - Penton, D.* AU - Abate, A.* AU - Peitzsch, M.* AU - Cosentini, D.* AU - Tiberio, G.A.M.* AU - Bornstein, S.R. AU - Berruti, A.* AU - Hantel, C.* C1 - 64914 C2 - 52529 TI - A comprehensive investigation of steroidogenic signaling in classical and new experimental cell models of adrenocortical carcinoma. JO - Cells VL - 11 IS - 9 PY - 2022 SN - 2073-4409 ER - TY - JOUR AB - Endolysosomal cation channels are emerging as key players of endolysosomal function such as endolysosomal trafficking, fusion/fission, lysosomal pH regulation, autophagy, lysosomal exocytosis, and endocytosis. Diseases comprise lysosomal storage disorders (LSDs) and neurode-generative diseases, metabolic diseases, pigmentation defects, cancer, immune disorders, autophagy related diseases, infectious diseases and many more. Involvement in lung diseases has not been a focus of attention so far but recent developments in the field suggest critical functions in lung physiology and pathophysiology. Thus, loss of TRPML3 was discovered to exacerbate emphysema formation and cigarette smoke induced COPD due to dysregulated matrix metalloproteinase 12 (MMP-12) levels in the extracellular matrix of the lung, a known risk factor for emphysema/COPD. While direct lung function measurements with the exception of TRPML3 are missing for other endolysosomal cation channels or channels expressed in lysosome related organelles (LRO) in the lung, links between those channels and important roles in lung physiology have been established such as the role of P2X4 in surfactant release from alveolar epithelial Type II cells. Other channels with demonstrated functions and disease relevance in the lung such as TRPM2, TRPV2, or TRPA1 may mediate their effects due to plasma membrane expression but evidence accumulates that these channels might also be expressed in endolysosomes, suggesting additional and/or dual roles of these channels in cell and intracellular membranes. We will discuss here the current knowledge on cation channels residing in endolysosomes or LROs with respect to their emerging roles in lung disease. AU - Spix, B.* AU - Jeridi, A. AU - Ansari, M. AU - Yildirim, A.Ö. AU - Schiller, H. B. AU - Grimm, C.* C1 - 64113 C2 - 52082 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Endolysosomal cation channels and lung disease. JO - Cells VL - 11 IS - 2 PB - Mdpi PY - 2022 SN - 2073-4409 ER - TY - JOUR AB - Smarca5, an ATPase of the ISWI class of chromatin remodelers, is a key regulator of chromatin structure, cell cycle and DNA repair. Smarca5 is deregulated in leukemia and breast, lung and gastric cancers. However, its role in oncogenesis is not well understood. Chromatin remodelers often play dosage-dependent roles in cancer. We therefore investigated the epigenomic and phenotypic impact of controlled stepwise attenuation of Smarca5 function in the context of primary cell transformation, a process relevant to tumor formation. Upon conditional single-or double-allele Smarca5 deletion, the cells underwent both accelerated growth arrest and senescence entry and displayed gradually increased sensitivity to genotoxic insults. These phenotypic characteristics were explained by specific remodeling of the chromatin structure and the transcriptome in primary cells prior to the immortalization onset. These molecular programs implicated Smarca5 requirement in DNA damage repair, telomere maintenance, cell cycle progression and in restricting apoptosis and cellular senescence. Consistent with the molecular programs, we demonstrate for the first time that Smarca5-deficient primary cells exhibit dramatically decreased capacity to bypass senescence and immortalize, an indispensable step during cell transformation and cancer development. Thus, Smarca5 plays a crucial role in key homeostatic processes and sustains cancer-promoting molecular programs and cellular phenotypes. AU - Thakur, S.* AU - Cahais, V.* AU - Turkova, T.* AU - Zikmund, T. AU - Renard, C.* AU - Stopka, T.* AU - Korenjak, M.* AU - Zavadil, J.* C1 - 64463 C2 - 52224 TI - Chromatin remodeler smarca5 is required for cancer-related processes of primary cell fitness and immortalization. JO - Cells VL - 11 IS - 5 PY - 2022 SN - 2073-4409 ER - TY - JOUR AB - Approximately 70 million humans worldwide are affected by chronic hepatitis D, which rapidly leads to liver cirrhosis and hepatocellular carcinoma due to chronic inflammation. The triggers and consequences of this chronic inflammation, induced by co-infection with the hepatitis D virus (HDV) and the hepatitis B virus (HBV), are poorly understood. Using CRISPR technology, we characterized the recognition of HDV mono-and co-infection by intracellular innate immunity and determined its influence on the viral life cycle and effector T-cell responses using different HBV and HDV permissive hepatoma cell lines. We showed that HDV infection is detected by MDA5 and-after a lag phase-induces a profound type I interferon response in the infected cells. The type I interferon response, however, was not able to suppress HDV replication or spread, thus providing a persistent trigger. Using engineered T-cells directed against the envelope proteins commonly used by HBV and HDV, we found that HDV immune recognition enhanced T-cell cytotoxicity. Interestingly, the T-cell effector function was enhanced independently of antigen presentation. These findings help to explain immune mediated tissue damage in chronic hepatitis D patients and indicate that combining innate triggers with T-cell activating therapies might allow for a curative approach. AU - Altstetter, S. AU - Quitt, O. AU - Pinci, F.* AU - Hornung, V.* AU - Lucko, A.M. AU - Wisskirchen, K. AU - Jung, S. AU - Protzer, U. C1 - 63563 C2 - 51588 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Hepatitis-D virus infection is not impaired by innate immunity but increases cytotoxic T-cell activity. JO - Cells VL - 10 IS - 11 PB - Mdpi PY - 2021 SN - 2073-4409 ER - TY - JOUR AB - Physical training improves insulin sensitivity and can prevent type 2 diabetes (T2D). However, approximately 20% of individuals lack a beneficial outcome in glycemic control. TGF-β, identified as a possible upstream regulator involved in this low response, is also a potent regulator of microRNAs (miRNAs). The aim of this study was to elucidate the potential impact of TGF-β-driven miRNAs on individual exercise response. Non-targeted long and sncRNA sequencing analyses of TGF-β1-treated human skeletal muscle cells corroborated the effects of TGF-β1 on muscle cell differentiation, the induction of extracellular matrix components, and identified several TGF-β1-regulated miRNAs. qPCR validated a potent upregulation of miR-143-3p/145-5p and miR-181a2-5p by TGF-β1 in both human myoblasts and differentiated myotubes. Healthy subjects who were overweight or obese participated in a supervised 8-week endurance training intervention (n = 40) and were categorized as responder or low responder in glycemic control based on fold change ISIMats (≥+1.1 or <+1.1, respectively). In skeletal muscle biopsies of low responders, TGF-β signaling and miR-143/145 cluster levels were induced by training at much higher rates than among responders. Target-mining revealed HDACs, MYHs, and insulin signaling components INSR and IRS1 as potential miR-143/145 cluster targets. All these targets were down-regulated in TGF-β1-treated myotubes. Transfection of miR-143-3p/145-5p mimics in differentiated myotubes validated MYH1, MYH4, and IRS1 as miR-143/145 cluster targets. Elevated TGF-β signaling and miR-143/145 cluster induction in skeletal muscle of low responders might obstruct improvements in insulin sensitivity by training in two ways: by a negative impact of miR-143-3p on muscle cell fusion and myofiber functionality and by directly impairing insulin signaling via a reduction in INSR by TGF-β and finetuned IRS1 suppression by miR-143-3p. AU - Dreher, S.I.* AU - Höckele, S. AU - Huypens, P. AU - Irmler, M. AU - Hoffmann, C.* AU - Jeske, T. AU - Hastreiter, M. AU - Moller, A. AU - Birkenfeld, A.L. AU - Häring, H.-U. AU - Peter, A. AU - Beckers, J. AU - Hrabě de Angelis, M. AU - Weigert, C. C1 - 63763 C2 - 51619 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Tgf-β induction of mir-143/145 is associated to exercise response by influencing differentiation and insulin signaling molecules in human skeletal muscle. JO - Cells VL - 10 IS - 12 PB - Mdpi PY - 2021 SN - 2073-4409 ER - TY - JOUR AB - Ewing sarcoma (EwS) is an aggressive pediatric cancer of bone and soft tissues characterized by scant T cell infiltration and predominance of immunosuppressive myeloid cells. Given the important roles of extracellular vesicles (EVs) in cancer-host crosstalk, we hypothesized that EVs secreted by EwS tumors target myeloid cells and promote immunosuppressive phenotypes. Here, EVs were purified from EwS and fibroblast cell lines and exhibited characteristics of small EVs, including size (100-170 nm) and exosome markers CD63, CD81, and TSG101. Treatment of healthy donor-derived CD33+ and CD14+ myeloid cells with EwS EVs but not with fibroblast EVs induced pro-inflammatory cytokine release, including IL-6, IL-8, and TNF. Furthermore, EwS EVs impaired differentiation of these cells towards monocytic-derived dendritic cells (moDCs), as evidenced by reduced expression of co-stimulatory molecules CD80, CD86 and HLA-DR. Whole transcriptome analysis revealed activation of gene expression programs associated with immunosuppressive phenotypes and pro-inflammatory responses. Functionally, moDCs differentiated in the presence of EwS EVs inhibited CD4+ and CD8+ T cell proliferation as well as IFNγ release, while inducing secretion of IL-10 and IL-6. Therefore, EwS EVs may promote a local and systemic pro-inflammatory environment and weaken adaptive immunity by impairing the differentiation and function of antigen-presenting cells. AU - Gassmann, H.* AU - Schneider, K.* AU - Evdokimova, V.* AU - Ruzanov, P.* AU - Schober, S.J.* AU - Xue, B.* AU - von Heyking, K.* AU - Thiede, M.* AU - Richter, G.H.S.* AU - Pfaffl, M.W.* AU - Nössner, E. AU - Stein, L.D.* AU - Sorensen, P.H.B.* AU - Thiel, U.* C1 - 62874 C2 - 51124 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Ewing sarcoma-derived extracellular vesicles impair dendritic cell maturation and function. JO - Cells VL - 10 IS - 8 PB - Mdpi PY - 2021 SN - 2073-4409 ER - TY - JOUR AB - Dysfunction of the immunoproteasome has been implicated in cardiovascular and pulmonary diseases. Its potential as a biomarker for predicting disease stages, however, has not been investigated so far and population-based analyses on the impact of sex and age are missing. We here analyzed the activity of all six catalytic sites of the proteasome in isolated peripheral blood mononuclear cells obtained from 873 study participants of the KORA FF4 study using activity-based probes. The activity of the immuno-and standard proteasome correlated clearly with elevated leukocyte counts of study participants. Unexpectedly, we observed a strong sex dimorphism for proteasome activity with significantly lower immunoproteasome activity in women. In aging, almost all catalytic activities of the proteasome were activated in aged women while maintained upon aging in men. We also noted distinct sex-related activation patterns of standard and immunoproteasome active sites in chronic inflammatory diseases such as diabetes, cardiovascular diseases, asthma, or chronic obstructive pulmonary disease as determined by multiple linear regression modeling. Our data thus provides a conceptual framework for future analysis of immunoproteasome function as a bio-marker for chronic inflammatory disease development and progression. AU - Kammerl, I.E. AU - Flexeder, C. AU - Karrasch, S. AU - Thorand, B. AU - Heier, M. AU - Peters, A. AU - Schulz, H. AU - Meiners, S. C1 - 63698 C2 - 51614 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Blood immunoproteasome activity is regulated by sex, age and in chronic inflammatory diseases: A first population-based study. JO - Cells VL - 10 IS - 12 PB - Mdpi PY - 2021 SN - 2073-4409 ER - TY - JOUR AB - Biallelic pathogenic variants in CLPP, encoding mitochondrial matrix peptidase ClpP, cause a rare autosomal recessive condition, Perrault syndrome type 3 (PRLTS3). It is characterized by primary ovarian insufficiency and early sensorineural hearing loss, often associated with pro-gressive neurological deficits. Mouse models showed that accumulations of (i) its main protein in-teractor, the substrate-selecting AAA+ ATPase ClpX, (ii) mitoribosomes, and (iii) mtDNA nucleoids are the main cellular consequences of ClpP absence. However, the sequence of these events and their validity in human remain unclear. Here, we studied global proteome profiles to define ClpP substrates among mitochondrial ClpX interactors, which accumulated consistently in ClpP-null mouse embryonal fibroblasts and brains. Validation work included novel ClpP-mutant patient fi-broblast proteomics. ClpX co-accumulated in mitochondria with the nucleoid component POLDIP2, the mitochondrial poly(A) mRNA granule element LRPPRC, and tRNA processing factor GFM1 (in mouse, also GRSF1). Only in mouse did accumulated ClpX, GFM1, and GRSF1 appear in nuclear fractions. Mitoribosomal accumulation was minor. Consistent accumulations in murine and human fibroblasts also affected multimerizing factors not known as ClpX interactors, namely, OAT, ASS1, ACADVL, STOM, PRDX3, PC, MUT, ALDH2, PMPCB, UQCRC2, and ACADSB, but the impact on downstream metabolites was marginal. Our data demonstrate the primary impact of ClpXP on the assembly of proteins with nucleic acids and show nucleoid enlargement in human as a key consequence. AU - Key, J.* AU - Torres-Odio, S.* AU - Bach, N.C.* AU - Gispert, S.* AU - Koepf, G.* AU - Reichlmeir, M.* AU - West, A.P.* AU - Prokisch, H. AU - Freisinger, P.* AU - Newman, W.G.* AU - Shalev, S.* AU - Sieber, S.A.* AU - Wittig, I.* AU - Auburger, G.* C1 - 63697 C2 - 51613 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Inactivity of peptidase clpp causes primary accumulation of mitochondrial disaggregase clpx with its interacting nucleoid proteins, and of mtdna. JO - Cells VL - 10 IS - 12 PB - Mdpi PY - 2021 SN - 2073-4409 ER - TY - JOUR AB - Retinal Müller glial cells (RMG) are involved in virtually every retinal disease; however, the role of these glial cells in neuroinflammation is still poorly understood. Since cell surface proteins play a decisive role in immune system signaling pathways, this study aimed at characterizing the changes of the cell surface proteome of RMG after incubation with prototype immune system stimulant lipopolysaccharide (LPS). While mass spectrometric analysis of the human Müller glia cell line MIO-M1 revealed 507 cell surface proteins in total, with 18 proteins significantly more abundant after stimulation (ratio ≥ 2), the surfaceome of primary RMG comprised 1425 proteins, among them 79 proteins with significantly higher abundance in the stimulated state. Pathway analysis revealed notable association with immune system pathways such as "antigen presentation", "immunoregulatory interactions between a lymphoid and a non-lymphoid cell" and "cell migration". We could demonstrate a higher abundance of proteins that are usually ascribed to antigen-presenting cells (APCs) and function to interact with T-cells, suggesting that activated RMG might act as atypical APCs in the course of ocular neuroinflammation. Our data provide a detailed description of the unstimulated and stimulated RMG surfaceome and offer fundamental insights regarding the capacity of RMG to actively participate in neuroinflammation in the retina. AU - Lorenz, L.* AU - Hirmer, S.* AU - Schmalen, A. AU - Hauck, S.M. AU - Deeg, C.A.* C1 - 61740 C2 - 50439 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Cell surface profiling of retinal Müller glial cells reveals association to immune pathways after LPS stimulation. JO - Cells VL - 10 IS - 3 PB - Mdpi PY - 2021 SN - 2073-4409 ER - TY - JOUR AB - Background: Although several approaches have revealed much about individual factors that regulate pancreatic development, we have yet to fully understand their complicated interplay during pancreas morphogenesis. Gfi1 is transcription factor specifically expressed in pancreatic acinar cells, whose role in pancreas cells fate identity and specification is still elusive. Methods: In order to gain further insight into the function of this factor in the pancreas, we generated animals deficient for Gfi1 specifically in the pancreas. Gfi1 conditional knockout animals were phenotypically characterized by immunohistochemistry, RT-qPCR, and RNA scope. To assess the role of Gfi1 in the pathogenesis of diabetes, we challenged Gfi1-deficient mice with two models of induced hyperglycemia: long-term high-fat/high-sugar feeding and streptozotocin injections. Results: Interestingly, mutant mice did not show any obvious deleterious phenotype. However, in depth analyses demonstrated a significant decrease in pancreatic amylase expression, leading to a diminution in intestinal carbohydrates pro-cessing and thus glucose absorption. In fact, Gfi1-deficient mice were found resistant to diet-induced hyperglycemia, appearing normoglycemic even after long-term high-fat/high-sugar diet. Another feature observed in mutant acinar cells was the misexpression of ghrelin, a hormone previously sug-gested to exhibit anti-apoptotic effects on β-cells in vitro. Impressively, Gfi1 mutant mice were found to be resistant to the cytotoxic and diabetogenic effects of high-dose streptozotocin administrations, displaying a negligible loss of β-cells and an imperturbable normoglycemia. Conclusions: Together, these results demonstrate that Gfi1 could turn to be extremely valuable for the development of new therapies and could thus open new research avenues in the context of diabetes research. AU - Napolitano, T.* AU - Avolio, F.* AU - Silvano, S.* AU - Forcisi, S. AU - Pfeifer, A.* AU - Vieira, A.* AU - Navarro-Sanz, S.* AU - Friano, M.E.* AU - Ayachi, C.* AU - Garrido-Utrilla, A.* AU - Atlija, J.* AU - Hadzic, B.* AU - Becam, J.* AU - Sousa-De-veiga, A.* AU - Plaisant, M.D.* AU - Balaji, S.* AU - Pisani, D.F.* AU - Mondin, M.* AU - Schmitt-Kopplin, P. AU - Amri, E.Z.* AU - Collombat, P.* C1 - 63327 C2 - 51472 TI - Gfi1 loss protects against two models of induced diabetes. JO - Cells VL - 10 IS - 11 PY - 2021 SN - 2073-4409 ER - TY - JOUR AB - Hidradenitis suppurativa (HS) is a chronic inflammatory skin disease primarily affecting apocrine gland-rich areas of the body. It is a multifactorial disease in which genetic and environmental factors play a key role. The primary defect in HS pathophysiology involves follicular occlusion of the folliculopilosebaceous unit, followed by follicular rupture and immune responses. Innate pro-inflammatory cytokines (e.g., IL-1β, and TNF-α); mediators of activated T helper (Th)1 and Th17 cells (e.g., IFN-γ, and IL-17); and effector mechanisms of neutrophilic granulocytes, macrophages, and plasma cells are involved. On the other hand, HS lesions contain anti-inflammatory mediators (e.g., IL-10) and show limited activity of Th22 cells. The inflammatory vicious circle finally results in pain, purulence, tissue destruction, and scarring. HS pathogenesis is still enigmatic, and a valid animal model for HS is currently not available. All these aspects represent a challenge for the development of therapeutic approaches, which are urgently needed for this debilitating disease. Available treatments are limited, mostly off-label, and surgical interventions are often required to achieve remission. In this paper, we provide an overview of the current knowledge surrounding HS, including the diagnosis, pathogenesis, treatments, and existing translational studies. AU - Scala, E.* AU - Cacciapuoti, S.* AU - Garzorz-Stark, N.* AU - Megna, M.* AU - Marasca, C.* AU - Seiringer, P. AU - Volz, T.* AU - Eyerich, K.* AU - Fabbrocini, G.* C1 - 62872 C2 - 51121 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Hidradenitis suppurativa: Where we are and where we are going. JO - Cells VL - 10 IS - 8 PB - Mdpi PY - 2021 SN - 2073-4409 ER - TY - JOUR AB - Whilst the importance of keratinocytes as a first-line defense has been widely investigated, little is known about their interactions with non-resident immune cells. In this study, the impact of human keratinocytes on T cell effector functions was analyzed in an antigen-specific in vitro model of allergic contact dermatitis (ACD) to nickel sulfate. Keratinocytes partially inhibited T cell proliferation and cytokine production. This effect was dependent on the keratinocyte/T cell ratio and was partially reversible by increasing the number of autologous dendritic cells. The inhibition of T cell proliferation by keratinocytes was independent of the T cell subtype and antigen presentation by different professional antigen-presenting cells. Autologous and heterologous keratinocytes showed comparable effects, while the fixation of keratinocytes with paraformaldehyde abrogated the immunosuppressive effect. The separation of keratinocytes and T cells by a transwell chamber, as well as a cell-free keratinocyte supernatant, inhibited T cell effector functions to the same amount as directly co-cultured keratinocytes, thus proving that soluble factor/s account for the observed suppressive effects. In conclusion, keratinocytes critically control the threshold of inflammatory processes in the skin by inhibiting T cell proliferation and cytokine production. AU - Seiringer, P. AU - Eyerich, S. AU - Eyerich, K. AU - Dittlein, D. AU - Pilz, A.C. AU - Scala, E.* AU - Ring, J.* AU - Behrendt, H. AU - Cavani, A.* AU - Traidl-Hoffmann, C. C1 - 62436 C2 - 50869 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Keratinocytes regulate the threshold of inflammation by inhibiting T cell effector functions. JO - Cells VL - 10 IS - 7 PB - Mdpi PY - 2021 SN - 2073-4409 ER - TY - JOUR AB - The heart tissue is a potential target of various noxae contributing to the onset of cardiovascular diseases. However, underlying pathophysiological mechanisms are largely unknown. Human stem cell-derived models are promising, but a major concern is cell immaturity when estimating risks for adults. In this study, 3D aggregates of human embryonic stem cell-derived cardiomyocytes were cultivated for 300 days and characterized regarding degree of maturity, structure, and cell composition. Furthermore, effects of ionizing radiation (X-rays, 0.1–2 Gy) on matured aggregates were investigated, representing one of the noxae that are challenging to assess. Video-based functional analyses were correlated to changes in the proteome after irradiation. Cardiomyocytes reached maximum maturity after 100 days in cultivation, judged by α-actinin lengths, and displayed typical multinucleation and branching. At this time, aggregates contained all major cardiac cell types, proven by the patch-clamp technique. Matured and X-ray-irradiated aggregates revealed a subtle increase in beat rates and a more arrhythmic sequence of cellular depolarisation and repolarisation compared to non-irradiated sham controls. The proteome analysis provides first insights into signaling mechanisms contributing to cardiotoxicity. Here, we propose an in vitro model suitable to screen various noxae to target adult cardiotoxicity by preserving all the benefits of a 3D tissue culture. AU - Smit, T.* AU - Schickel, E.* AU - Azimzadeh, O. AU - von Toerne, C. AU - Rauh, O.* AU - Ritter, S.* AU - Durante, M.* AU - Schroeder, I.S.* C1 - 63204 C2 - 51383 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - A human 3d cardiomyocyte risk model to study the cardiotoxic influence of x-rays and other noxae in adults. JO - Cells VL - 10 IS - 10 PB - Mdpi PY - 2021 SN - 2073-4409 ER - TY - JOUR AB - 5 ' AMP-activated protein kinase (AMPK) is known as metabolic sensor in mammalian cells that becomes activated by an increasing adenosine monophosphate (AMP)/adenosine triphosphate (ATP) ratio. The heterotrimeric AMPK protein comprises three subunits, each of which has multiple phosphorylation sites, playing an important role in the regulation of essential molecular pathways. By phosphorylation of downstream proteins and modulation of gene transcription AMPK functions as a master switch of energy homeostasis in tissues with high metabolic turnover, such as the liver, skeletal muscle, and adipose tissue. Regulation of AMPK under conditions of chronic caloric oversupply emerged as substantial research target to get deeper insight into the pathogenesis of non-alcoholic fatty liver disease (NAFLD). Evidence supporting the role of AMPK in NAFLD is mainly derived from preclinical cell culture and animal studies. Dysbalanced de novo lipogenesis has been identified as one of the key processes in NAFLD pathogenesis. Thus, the scope of this review is to provide an integrative overview of evidence, in particular from clinical studies and human samples, on the role of AMPK in the regulation of primarily de novo lipogenesis in human NAFLD. AU - von Loeffelholz, C.* AU - Coldewey, S.M.* AU - Birkenfeld, A.L. C1 - 62728 C2 - 51027 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - A narrative review on the role of AMPK on de novo lipogenesis in non-alcoholic fatty liver disease: Evidence from human studies. JO - Cells VL - 10 IS - 7 PB - Mdpi PY - 2021 SN - 2073-4409 ER - TY - JOUR AB - Chronic hepatitis C virus (HCV) infection is closely associated with a plethora of diseases, including cancers and autoimmune disorders. However, the distinct triggers and cellular networks leading to such HCV-derived diseases are poorly understood. Around 8% of the human genome consists of human endogenous retroviruses. They are usually silenced but can be reactivated by environmental conditions, including viral infections. Our current understanding indicates that the activation of one specific family-namely, HERV-K(HML-2)-is linked to distinct pathologies, including cancer and autoimmunity. In this study, we analyzed the transcription levels of HERV-K(HML-2) in 42 HCV-infected patients receiving direct-acting antiviral therapies. Samples from the start of treatment until 12 weeks post-treatment were investigated. Our results show increased HERV-K(HML-2) transcript levels in patients with HCV-derived liver cirrhosis throughout the observation period. Several clinical parameters specifying poor liver function are positively correlated with HERV-K(HML-2) expression. Of note, patients without a sustained viral clearance showed a drastic increase in HERV-K(HML-2) transcript levels. Together, our data suggest that increased HERV-K(HML-2) expression is correlated with reduced liver function as well as therapy success in HCV-infected patients. AU - Weber, M. AU - Padmanabhan Nair, V. AU - Bauer, T. AU - Sprinzl, M.F.* AU - Protzer, U. AU - Vincendeau, M. C1 - 61738 C2 - 50438 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Increased HERV-K(HML-2) transcript levels correlate with clinical parameters of liver damage in hepatitis C patients. JO - Cells VL - 10 IS - 4 PB - Mdpi PY - 2021 SN - 2073-4409 ER - TY - JOUR AB - Osteoarthritis (OA) is a degenerative disease of the hyaline articular cartilage. This disease is progressive and may lead to disability. Researchers proposed many regenerative approaches to treat osteoarthritis, including stem cells. Trans-differentiation of a fully differentiated cell state directly into another different differentiated cell state avoids the disadvantages of fully reprogramming cells to induced pluripotent stem cells (iPSCs) in terms of faster reprogramming of the needed cells. Trans-differentiation also reduces the risk of tumor formation by avoiding the iPSC state. OSKM factors (Oct4, Sox2, Klf4, and cMyc) accompanied by the JAK-STAT pathway inhibition, followed by the introduction of specific differentiation factors, directly reprogrammed mouse embryonic fibroblasts to chondroblasts. Our results showed the absence of intermediate induced pluripotent stem cell formation. The resulting aggregates showed clear hyaline and hypertrophic cartilage. Tumor formation was absent in sub-cutaneous capsules transplanted in SCID mice. AU - Cota, P. AU - Helmi, S.A.* AU - Hsu, C.* AU - Rancourt, D.E.* C1 - 57855 C2 - 48162 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Cytokine directed chondroblast trans-differentiation: JAK inhibition facilitates direct reprogramming of fibroblasts to chondroblasts. JO - Cells VL - 9 IS - 1 PB - Mdpi PY - 2020 SN - 2073-4409 ER - TY - JOUR AB - The identification of human obesity sub-types may improve the clinical management of patients with obesity and uncover previously unrecognized obesity mechanisms. Here, we hypothesized that adipose tissue (AT) mast cells (MC) estimation could be a mark for human obesity sub-phenotyping beyond current clinical-based stratifications, both cross-sectionally and prospectively. We estimated MC accumulation using immunohistochemistry and gene expression in abdominal visceral AT (VAT) and subcutaneous (SAT) in a human cohort of 65 persons with obesity who underwent elective abdominal (mainly bariatric) surgery, and we validated key results in two clinically similar, independent cohorts (n= 33,n= 56). AT-MC were readily detectable by immunostaining for either c-kit or tryptase and by assessing the gene expression of KIT (KIT Proto-Oncogene, Receptor Tyrosine Kinase), TPSB2 (tryptase beta 2), and CMA1 (chymase 1). Participants were characterized as VAT-MC(low)if the expression of both CMA1 and TPSB2 was below the median. Higher expressers of MC genes (MChigh) were metabolically healthier (lower fasting glucose and glycated hemoglobin, with higher pancreatic beta cell reserve (HOMA-beta), and lower triglycerides and alkaline-phosphatase) than people with low expression (MClow). Prospectively, higher MC accumulation in VAT or SAT obtained during surgery predicted greater postoperative weight-loss response to bariatric surgery. Jointly, high AT-MC accumulation may be used to clinically define obesity sub-phenotypes, which are associated with a "healthier" cardiometabolic risk profile and a better weight-loss response to bariatric surgery. AU - Goldstein, N.* AU - Kezerle, Y.* AU - Gepner, Y.* AU - Haim, Y.* AU - Pecht, T.* AU - Gazit, R.* AU - Polischuk, V.* AU - Liberty, I.F.* AU - Kirshtein, B.* AU - Shaco-Levy, R.* AU - Blüher, M. AU - Rudich, A.* C1 - 59479 C2 - 48880 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Higher mast cell accumulation in human adipose tissues defines clinically favorable obesity sub-phenotypes. JO - Cells VL - 9 IS - 6 PB - Mdpi PY - 2020 SN - 2073-4409 ER - TY - JOUR AB - The ongoing threat of viral infections and the emergence of antiviral drug resistance warrants a ceaseless search for new antiviral compounds. Broadly-inhibiting compounds that act on elements shared by many viruses are promising antiviral candidates. Here, we identify a peptide derived from the cowpox virus protein CPXV012 as a broad-spectrum antiviral peptide. We found that CPXV012 peptide hampers infection by a multitude of clinically and economically important enveloped viruses, including poxviruses, herpes simplex virus-1, hepatitis B virus, HIV-1, and Rift Valley fever virus. Infections with non-enveloped viruses such as Coxsackie B3 virus and adenovirus are not affected. The results furthermore suggest that viral particles are neutralized by direct interactions with CPXV012 peptide and that this cationic peptide may specifically bind to and disrupt membranes composed of the anionic phospholipid phosphatidylserine, an important component of many viral membranes. The combined results strongly suggest that CPXV012 peptide inhibits virus infections by direct interactions with phosphatidylserine in the viral envelope. These results reiterate the potential of cationic peptides as broadly-acting virus inhibitors. AU - Luteijn, R.D.* AU - Praest, P.* AU - Thiele, F. AU - Sadasivam, S.M.* AU - Singethan, K. AU - Drijfhout, J.W.* AU - Bach, C.E. AU - de Boer, S.M.* AU - Lebbink, R.J.* AU - Tao, S.* AU - Helfer, M.* AU - Bach, N.C.* AU - Protzer, U. AU - Costa, A.I.* AU - Killian, J.A.* AU - Drexler, I.* AU - Wiertz, E.J.H.J.* C1 - 60010 C2 - 49191 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - A broad-spectrum antiviral peptide blocks infection of viruses by binding to phosphatidylserine in the viral envelope. JO - Cells VL - 9 IS - 9 PB - Mdpi PY - 2020 SN - 2073-4409 ER - TY - JOUR AB - Granulins (GRN) are secreted factors that promote neuronal survival and regulate inflammation in various pathological conditions. However, their roles in physiological conditions in the brain remain poorly understood. To address this knowledge gap, we analysed the telencephalon in Grn-deficient zebrafish and identified morphological and transcriptional changes in microglial cells, indicative of a pro-inflammatory phenotype in the absence of any insult. Unexpectedly, activated mutant microglia shared part of their transcriptional signature with aged human microglia. Furthermore, transcriptome profiles of the entire telencephali isolated from young Grn-deficient animals showed remarkable similarities with the profiles of the telencephali isolated from aged wildtype animals. Additionally, 50% of differentially regulated genes during aging were regulated in the telencephalon of young Grn-deficient animals compared to their wildtype littermates. Importantly, the telencephalon transcriptome in young Grn-deficent animals changed only mildly with aging, further suggesting premature aging of Grn-deficient brain. Indeed, Grn loss led to decreased neurogenesis and oligodendrogenesis, and to shortening of telomeres at young ages, to an extent comparable to that observed during aging. Altogether, our data demonstrate a role of Grn in regulating aging kinetics in the zebrafish telencephalon, thus providing a valuable tool for the development of new therapeutic approaches to treat age-associated pathologies. AU - Zambusi, A. AU - Pelin Burhan, Ö.* AU - Di Giaimo, R. AU - Schmid, B. AU - Ninkovic, J. C1 - 58110 C2 - 48197 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Granulins regulate aging kinetics in the adult zebrafish telencephalon. JO - Cells VL - 9 IS - 2 PB - Mdpi PY - 2020 SN - 2073-4409 ER - TY - JOUR AB - The inhibition of heat shock protein 90 (Hsp90) a molecular chaperone for multiple oncogenic client proteins is considered as a promising approach to overcome radioresistance. Since most Hsp90 inhibitors activate HSF-1 that induces the transcription of cytoprotective and tumor-promoting stress proteins such as Hsp70 and Hsp27, a combined approach consisting of HSF-1 knockdown (k.d.) and Hsp90 inhibition was investigated. A specific HSF-1 k.d. was achieved in H1339 lung cancer cells using RNAi-Ready pSIRENRetroQ vectors with puromycin resistance. The Hsp90 inhibitor NVP-AUY922 was evaluated at low concentrations-ranging from 1-10 nM-in control and HSF-1 k.d. cells. Protein expression (i.e., Hsp27/Hsp70, HSF-1, pHSF-1, Akt, beta-actin) and transcriptional activity was assessed by western blot analysis and luciferase assays and radiosensitivity was measured by proliferation, apoptosis (Annexin V, active caspase 3), clonogenic cell survival, alkaline comet, gamma H2AX, 53BP1, and Rad51 foci assays. The k.d. of HSF-1 resulted in a significant reduction of basal and NVP-AUY922-induced Hsp70/Hsp27 expression levels. A combined approach consisting of HSF-1 k.d. and low concentrations of the Hsp90 inhibitor NVP-AUY922 reduces the Hsp90 client protein Akt and potentiates radiosensitization, which involves an impaired homologous recombination mediated by Rad51. Our findings are key for clinical applications of Hsp90 inhibitors with respect to adverse hepatotoxic effects. AU - Kühnel, A. AU - Schilling, D. AU - Combs, S.E. AU - Haller, B.* AU - Schwab, M.* AU - Multhoff, G.* C1 - 57003 C2 - 47429 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Radiosensitization of HSF-1 knockdown lung cancer cells by low concentrations of Hsp90 inhibitor NVP-AUY922. JO - Cells VL - 8 IS - 10 PB - Mdpi PY - 2019 SN - 2073-4409 ER - TY - JOUR AB - The glycoprotein osteopontin (OPN) possesses multiple functions in health and disease. To this end, osteopontin has beneficial roles in wound healing, bone homeostasis, and extracellular matrix (ECM) function. On the contrary, osteopontin can be deleterious for the human body during disease. Indeed, osteopontin is a cardinal mediator of tumor-associated inflammation and facilitates metastasis. The purpose of this review is to highlight the importance of osteopontin in malignant processes, focusing on lung and pleural tumors as examples. AU - Lamort, A.-S. AU - Giopanou, I.* AU - Psallidas, I.* AU - Stathopoulos, G.T. C1 - 56717 C2 - 47232 TI - Osteopontin as a link between inflammation and cancer: The thorax in the spotlight. JO - Cells VL - 8 IS - 8 PY - 2019 SN - 2073-4409 ER - TY - JOUR AB - Plants constantly suffer from simultaneous infection by multiple pathogens, which can be divided into biotrophic, hemibiotrophic, and necrotrophic pathogens, according to their lifestyles. Many studies have contributed to improving our knowledge of how plants can defend against pathogens, involving different layers of defense mechanisms. In this sense, the review discusses: (1) the functions of PAMP (pathogen-associated molecular pattern)-triggered immunity (PTI) and effector-triggered immunity (ETI), (2) evidence highlighting the functions of salicylic acid (SA) and jasmonic acid (JA)/ethylene (ET)-mediated signaling pathways downstream of PTI and ETI, and (3) other defense aspects, including many novel small molecules that are involved in defense and phenomena, including systemic acquired resistance (SAR) and priming. In particular, we mainly focus on SA and (JA)/ET-mediated signaling pathways. Interactions among them, including synergistic effects and antagonistic effects, are intensively explored. This might be critical to understanding dynamic disease regulation. AU - Zhang, W.* AU - Zhao, F. AU - Jiang, L.* AU - Chen, C.* AU - Wu, L.* AU - Liu, Z.* C1 - 54969 C2 - 45999 CY - St Alban-anlage 66, Ch-4052 Basel, Switzerland TI - Different pathogen defense strategies in Arabidopsis: More than pathogen recognition. JO - Cells VL - 7 IS - 12 PB - Mdpi PY - 2018 SN - 2073-4409 ER -