@phdthesis{Liedtke2023, author = {Liedtke, Victoria}, title = {Lens epithelium-derived growth factor (LEDGF) as an aid in the diagnosis of benign prostatic hyperplasia (BPH) and the molecular principles of LEDGF in human cancer cells}, doi = {10.26127/BTUOpen-6698}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:co1-opus4-66988}, school = {BTU Cottbus - Senftenberg}, year = {2023}, abstract = {Benign prostatic hyperplasia (BPH) is considered an age-related disease of men with unknown etiopathophysiology. Chronic inflammation represents one of the most important pathophysiological mechanisms in this context, and there is also evidence for the involvement of autoimmune responses in an inflammatory environment in the prostate. Conventional therapeutic approaches for BPH are limited; however, immunosuppression by TNF (tumor necrosis factor)-𝛼 blockers seems to reduce symptoms in BPH patients due to the reduction of epithelial hyperplasia and macrophage-triggered inflammation. Therefore, this cumulative dissertation addresses biomarker-based BPH diagnosis by lens epithelium-derived growth factor splice variant 75 kDa (LEDGF/p75). LEDGF/p75 is an autoantigen that is overexpressed in solid tumors and functions as a stress-related transcriptional co-activator. It also plays an important role in cancer and DNA damage repair (DDR). The generation of CRISPR/Cas9-modified HEp-2 LEDGF deficient (KO) and HEp-2 LEDGF/p75 overexpressing (OE) cell lines provided new insights for the diagnosis and therapy of cancer and genetic disorders. Herein, IgG autoantibody reactivity to LEDGF/p75 was detected 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) linear immunoassay (LIA) and Chritidia luciliae immunofluorescence test (CLIFT). In addition, proliferation and migration capacity as well as chemosensitivity of genome-edited cells were determined by optimized all-in-one approach, followed by investigation of DDR signaling pathways by Western blot and immunofluorescence. Surprisingly, no increased binding of autoantibodies (autoAbs) to LEDGF/p75 was detected in cancer patients, but autoAb reactivity to LEDGF/p75 O/E cells was unexpectedly increased in approximately 50\% of patients with BPH. At the molecular level, LEDGF-deficient cells exhibit decreased proliferation and migration and increased sensitivity to the chemotherapeutic agent etoposide. A significantly Decreased DNA damage response could also be visualized. New diagnostic capabilities using HEp-2 cells overexpressing LEDGF/p75 or mitochondrial DNA as autoimmune targets could be used to identify BPH patients with autoimmune responses. Furthermore, this Thesis shows that LEDGF is not only involved in the recruitment of DNA damage response proteins, but also affects the proteosomal regulation of DDR signaling molecules.}, subject = {Y-H2AX; LEDGF/p75; CRISPR/Cas9; Prognostic biomarker; DNA damage signaling; Autoimmunity; DNA Reparatur; Darmkrebs; Autoimmunerkrankungen; Autoaggressionskrankheit; Prostatahypertrophie; Autoantigen; DNS-Reparatur}, language = {en} }