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 - 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 - TY - GEN A1 - Weiss, Romano A1 - Liedtke, Victoria A1 - Rödiger, Stefan T1 - Cannabidiol does not cause DNA double-strand breaks in a human liver-derived cell model T2 - Journal of cannabis research N2 - Background Cannabidiol (CBD) is a non-psychoactive cannabinoid with potential therapeutic applications, including anti-inflammatory, analgesic, and anticancer effects. However, experts raised concerns about its potential to induce DNA damage and chromosomal aberrations at low concentrations. Notably, these studies used liver cell lines, which may not fully reflect the metabolic processing of CBD, potentially limiting the generalizability of their findings. This study investigated the short time effects of CBD on DNA double-strand breaks (DSBs) and proliferation in the human liver-derived cell line HepG2. Methods HepG2 cells were treated with CBD (5 – 50 , 3 - 72h incubation). To investigate potential imbalances in the expression of cannabinoid receptors 1 and 2 (CB1 / CB2) within HepG2 cells, we examined their expression using Western blot analysis. We hypothesized that such an imbalance could be associated with pathogenic processes. Double-strand breaks were then detected (5 Etoposide (ETP) served as positive control) via indirect immunofluorescence analysis using H2AX and 53BP1 antibodies, followed by quantification of DSB foci. Results Expression of CB2 but not CB1 was downregulated by 30 % in HepG2 cells after exposure to 5 CBD (24h incubation; 0.05) and 70 % downregulated after exposure to 50 CBD (24h incubation; 0.01). This effect was dose-dependent. Whilst ETP induced dose dependent DSBs, we could not confirm findings by others that CBD significantly increases the number of H2AX and 53BP1 foci between 5 and 50 (3h incubation; 0.05). Conclusion In our model, CBD stimulated the cells, as confirmed by modulation of CB2 expression as well as changes in intracellular cAMP. Our results show that CBD in ranges between 5 to 50 does not significantly increase the amount of DNA double strand breaks in HepG2 cells compared to the control. However, we did observe a significant reduction in cell proliferation and a significant increase in intracellular cAMP levels following CBD treatment. KW - Cannabidiol KW - DNA damage KW - Proliferation KW - CAMP KW - HepG2 Y1 - 2025 U6 - https://doi.org/10.1186/s42238-025-00365-w SN - 2522-5782 VL - 8 IS - 1 SP - 1 EP - 15 PB - Springer Science and Business Media LLC ER -