TY - GEN A1 - Lutter, Anne-Helen A1 - Liedtke, Victoria A1 - Ecke, Annemarie A1 - Scholka, Jenny A1 - Muschter, Antje A1 - Hentschel, Erik A1 - Becker, Roland A1 - Anderer, Ursula T1 - Direct co-culture of human chondrogenic microtissues with osteoblast-like Saos-2 cells positively influence the differentiation potential of cartilage cells in 3D culture Y1 - 2018 U6 - https://doi.org/10.1007%2Fs10354-018-0664-9.pdf N1 - Austrian Bone Conference – ABC 2018, November 23–24, 2018, Vienna, Austria ER - TY - GEN A1 - Lutter, Anne-Helen A1 - Liedtke, Victoria A1 - Scholka, Jenny A1 - Muschter, Antje A1 - Becker, Roland A1 - Anderer, Ursula T1 - Co-culture of human chondrogenic microtissues with osteoblast-like Saos-2 cells or HFF-1 fibroblasts influence the differentiation potential of spheroids T2 - Journal of Cellular Biotechnology N2 - BACKGROUND: Tissue engineering has become a major field of research in biotechnology and biomedicine. As a consequence, cell-based therapeutic approaches are entering the hospitals, especially for skeletal regeneration. Traumatic injuries of cartilage are treated with autologous cell suspensions or in vitro generated cartilage tissues, but there is actually no therapy available for degenerative cartilage defects. However, Osteoarthritis (OA) is a major public health problem in the world affecting 240 million people globally. OBJECTIVE: To develop suitable in vitro tissues, the properties of chondrogenic spheroids should be optimized via coculture with cells naturally occurring as joint neighbours. METHODS: Human chondrocytes were isolated from condyles and propagated in monolayer culture. Scaffold-free spheroids were generated and co-cultured with joint-specific partner cells (osteoblast-like osteosarcoma cells, fibroblasts). Morphology and differentiation was analyzed using histochemistry (Alcian blue, Safranin O) and immunohistochemistry for cartilage markers (collagen type II, Sox9, proteoglycan), proliferation-associated protein (Ki67) and markers of connective tissue (collagen type I and actin). RESULTS: The provision of a more natural microenvironment in vitro via co-culture of chondrocyte-based aggregates with osteoblast-like Saos-2 cells enhanced the differentiation potential of chondrogenic spheroids towards hyaline cartilage. CONCLUSIONS: The study showed the positive influence of Saos-2 cells on the differentiation potential of human chondrocytes in co-culture. Y1 - 2018 U6 - https://doi.org/10.3233/JCB-189005 SN - 2352-3697 SN - 2352-3689 VL - 4 IS - 1/2 SP - 31 EP - 42 ER - TY - GEN A1 - Liedtke, Victoria A1 - Schröder, Christian A1 - Roggenbuck, Dirk A1 - Weiss, Romano A1 - Stohwasser, Ralf A1 - Schierack, Peter A1 - Rödiger, Stefan A1 - Schenk, Lysann T1 - LEDGF/p75 is required for an efficient DNA damage response T2 - International Journal of Molecular Sciences KW - LEDGF KW - CRISPR/Cas9 KW - DNA damage signaling KW - gH2AX KW - ubiquitination Y1 - 2021 U6 - https://doi.org/10.3390/ijms22115866 VL - 22 IS - 11 SP - 1 EP - 16 ER - TY - GEN A1 - Liedtke, Victoria A1 - Stöckle, Michael A1 - Junker, Kerstin A1 - Roggenbuck, Dirk T1 - Benign prostatic hyperplasia - a novel autoimmune disease with a potential therapy consequence? T2 - Autoimmunity Reviews N2 - Benign prostatic hyperplasia (BPH) is considered as an age-related disease of men with an unknown etiopathophysiology. Chronic inflammation has been proposed as one of the major pathophysiological mechanisms. There is growing evidence for the involvement of autoimmune responses in an inflammatory setting in the prostate. Patients with autoimmune diseases show a significantly elevated prevalence of BPH. Conventional therapy options for BPH are limited, rendering surgery the ultimate alternative. However, immunosuppression via tumor necrosis factor alpha blocker appears to reduce symptoms in patients with BPH and concurrent autoimmune disease due to the reduction of epithelial hyperplasia and macrophage-induced inflammation. New diagnostic options using HEp-2 cells with overexpression of LEDGF/p75 or mitochondrial DNA as autoimmune targets could be used to identify BPH patients with autoimmune responses. Given the presumed involvement of autoimmune responses in BPH and the efficacy of immunosuppression in reducing BPH symptoms, BPH or subvariants of BPH may be candidates for a new autoimmune disease in males. KW - Autoimmunity Benign prostatic hyperplasia Anti-mitochondrial DNA antibodies LEDGF/p75 Y1 - 2024 U6 - https://doi.org/10.1016/j.autrev.2023.103511 SN - 1568-9972 VL - 23 IS - 3 PB - Elsevier BV ER - TY - GEN A1 - Liedtke, Victoria A1 - Weiss, Romano A1 - Skifov, Anastasia A1 - Rödiger, Stefan A1 - Schenk, Lysann T1 - Fully automated viability and toxicity screening - areliable all‐in‐one attempt T2 - Cancer Medicine N2 - The CRISPR/Cas9 technology is nowadays a common tool for genome editing to achieve new insights into, for example, diagnostics and therapeutics in cancer and genetic disorders. Cell proliferation and anticancer drug response studies are widely used to evaluate the impact of editing. However, these assays are often time‐consuming, expensive, and reproducibility is an issue. To overcome this, we developed a fast and cheap assay that combines a fully automated multispectral fluorescence microscopy platform with a nuclei staining and open‐source software analysis.Here, we generated different LEDGF/p75 model cell lines to validate the effect on proliferation and chemosensitivity. Therefore, a fast protocol for an optimized all‐in‐one attempt for cytotoxicity screenings and proliferation analysis of adherent cells in a 96‐well plate format was established using differential staining with two fluorescent dyes (Hoechst 33342 and propidium iodide) for live/dead cell discrimination. Subsequently, an automated cell nuclei count and analysis were performed using bioimage informatics.With the new established assay technology, up to 50,000 cells/well can be detected and analyzed in a 96‐well plate, resulting in a fast and accurate verification of viability and proliferation with consistency of 98% compared to manual counting. Our screening revealed that LEDGF depletion using CRISPR/Cas9 showed a diminished proliferation and chemosensitivity independent of cell line origin. Moreover, LEDGF depletion caused a significant increase in 𝛾H2AX foci, indicating a substantial increase in DNA double strand breaks. LEDGF/p75 overexpression enhanced proliferation and chemoresistance underlining the role of LEDGF in DNA damage response.Independent of cancer cell type, LEDGF/p75 is a central player in DNA damage repair and is implicated in chemoresistance. Moreover, our automated fluorescence biosensor technology allowed fast and reliable data acquisition without any fixation or additional washing steps. Additionally, data analysis was implemented using the modular open‐source software that can be adapted as needed. Y1 - 2024 U6 - https://doi.org/10.1002/cam4.7392 SN - 2045-7634 VL - 13 IS - 12 PB - Wiley ER - TY - GEN A1 - Liedtke, Victoria A1 - Rose, Laura A1 - Hiemann, Rico A1 - Nasser, Abdullah A1 - Rödiger, Stefan A1 - Bonaventura, Alena A1 - Winkler, Laura A1 - Sowa, Mandy A1 - Stöckle, Michael A1 - Schierack, Peter A1 - Junker, Kerstin A1 - Roggenbuck, Dirk T1 - Over-Expression of LEDGF/p75 in HEp-2 Cells Enhances Autoimmune IgG Response in Patients with Benign Prostatic Hyperplasia—A Novel Diagnostic Approach with Therapeutic Consequence? T2 - International Journal of Molecular Sciences N2 - Lens epithelium-derived growth factor splice variant of 75 kDa (LEDGF/p75) is an autoantigen over-expressed in solid tumors and acts as a stress-related transcriptional co-activator. Participation of autoimmune responses in the pathophysiology of benign prostatic hyperplasia (PBH) and a corresponding immunosuppressive therapy by TNFalpha antagonists has been recently suggested. Thus, autoAb testing could aid in the diagnosis of BPH patients profiting from such therapy. We generated CRISPR/Cas9 modified HEp-2 LEDGF knock-out (KO) and HEp-2 LEDGF/p75 over-expressing (OE) cells and examined IgG autoantibody reactivity to LEDGF/p75 in patients with prostate cancer (PCa, n = 89), bladder cancer (BCa, n = 116), benign prostatic hyperplasia (BPH, n = 103), and blood donors (BD, n = 60) by indirect immunofluorescence assay (IFA). Surprisingly, we could not detect elevated binding of autoAbs against LEDGF/p75 in cancer patients, but autoAb reactivity to LEDGF/p75 OE cells in about 50% of patients with BPH was unexpectedly significantly increased. Furthermore, a line immunoassay enabling the detection of 18 different autoAbs revealed a significantly increased occurrence of anti-dsDNA autoAbs in 34% of BPH patients in contrast to tumor patients and BD. This finding was confirmed by anti-mitochondrial (mDNA) autoAb detection with the Crithidia luciliae immunofluorescence test, which also showed a significantly higher prevalence (34%) of anti-mDNA autoAbs in BPH. In summary, our study provided further evidence for the occurrence of autoimmune responses in BPH. Furthermore, LEDGF/p75 over-expression renders HEp-2 cells more autoantigenic and an ideal target for autoAb analysis in BPH with a potential therapy consequence. KW - LEDGF/p75 KW - autoimmunity KW - CRISPR/Cas9 KW - dsDNA KW - mDNA Y1 - 2023 U6 - https://doi.org/10.3390/ijms24076166 SN - 1422-0067 VL - 24 IS - 7 ER -