Einfluss von Komplementrezeptor 2 Polymorphismen bei der Pathogenese des Systemischen Lupus Erythematosus

Diseases are termed genetic diseases if there exists a direct causal relationship between an identified genetic variation or mutation and the development of the respective clinical symptoms. An example for a genetic disease with known etiology is Cystic fibrosis. This disease is caused by a mutation within the gene for a chloride ion channel, resulting in production of tough secretions and re-occurring infections of the respiratory tracts. Thereby, 67% of all disease cases are caused by one defined mutation (deltaF508). Systemic Lupus Erythematosus (SLE, OMIM 152700) is a complex autoimmune disease showing a wide array of different organ manifestations. Despite the characteristical facial malar rash, SLE can also cause clinical manifestations in the central nervous system and the vasculatory system. The most severe organ manifestation is the glomerulonephritis of the kidney. Only in very rare cases, a causal effect could be identified, linking one specific genetic defect to the manifestation of clinical SLE symptoms. For the vast majority of cases, no single causal defect could be identified. The strong familiar aggregation of SLE cases strongly points to the fact, that genetic factors play an important role in the development of SLE but the complex pattern of inheritance and the incomplete penetrance even in monozygotic twins argue, that the development of SLE is caused by multiple, independent risk factors in combination with natural factors. The complement receptor 2 (CR2) has been shown to be a susceptibility gene for SLE in murine models of human SLE. Within this work, using a case-control association study, it was analyzed, if allelic variants of CR2 cause an elevated susceptibility for SLE or correlate with specific clinical manifestations. Using three independent populations of SLE patients and matched healthy control individuals, there was no evidence of association of the defined allelic variants of CR2 and SLE. Nevertheless, a correlation was discovered between the minor family of CR2 variants and an elevated age at the time point of the initial SLE diagnosis. Furthermore, there exists a trend, that the minor family of CR2 allelic variants could be correlated with a reduced risk of developing lupus nephritis. By analyzing human pre-B cells that have been transduced with the two main allelic variants of CR2, it was discovered, that the minor allelic variant of CD21 is unable to negatively regulate CD19 surface density. This result could be reproduced by analyzing the CD21 and CD19 cell surface density of freshly isolated primary human B cells from healthy donors. By assessing the total Ca2+ flux in CD21 transduced pre-B cells in a model of simultaneous cross-ligation of the pre-BCR and CR2, it was discovered, that the main allelic variants of CR2 differ in their capability to modulate the pre-BCR induced Ca2+ flux. In summary, the complement receptor 2 is a disease modifier for human SLE, with an allelic variant causing an elevated age at the initial diagnosis of SLE, an alteration in the ratio of CD19 vs. CD21 cell surface density on peripheral B cells and different signaling capabilities of pre-B cells with ectopic CD21 expression.Als genetisch bedingte Krankheiten werden diejenigen Krankheiten bezeichnet, bei denen ein direkter kausaler Zusammenhang zwischen einer genetischen Variation oder Mutation und der Ausprägung der entsprechenden klinischen Symptome besteht. Ein Beispiel für eine Erkrankung mit direkter genetischer Beteiligung ist die Zystische Fibrose. Bei dieser Erkrankung führt eine Mutation im Gen für einen Ionenkanal zur Ausscheidung von zähem Sekret und wiederkehrenden Infektionen der Atemwege. Dabei sind 67% aller Krankheitsfälle durch das Vorhandensein der häufigsten Mutation (deltaF508) zu erklären. Der Systemische Lupus Erythematosus (SLE, OMIM 152700) ist eine komplexe Autoimmunerkrankung, in deren Verlauf eine breite Zahl an Organbeteiligungen auftreten kann. Neben dem charakteristischen Schmetterlingserythem kann unter anderem auch das zentrale Nervensystem oder das Gefäßsystem betroffen sein. Die klinisch schwerwiegendste Organbeteiligung ist die Glomerulonephritis der Niere. Nur in seltenen Fällen konnte bisher ein direkter Zusammenhang zwischen einem einzelnen Gendefekt und der Manifestation von SLE Symptomen nachgewiesen werden. Für die überwiegende Mehrzahl aller Patienten kann jedoch kein kausaler Zusammenhang zwischen einem spezifischen Gendefekt und der Erkrankung definiert werden. Die bestätigte familiäre Häufung von SLE Fällen legt jedoch die Vermutung nahe, dass genetische Faktoren bei der Krankheitsentstehung maßgeblich beteiligt sind, jedoch deutet die komplexe Art der Vererbung und die unvollständige Penetranz bei eineiigen Zwillingen darauf hin, dass mehrere unabhängige Risikofaktoren mit jeweils nur einem geringen Krankheitsrisiko, sowie Umweltfaktoren die Krankheitsausprägung beeinflussen. Der Komplementrezeptor 2 (CR2) wurde in einem Maus Modell des humanen SLE als ein Suszeptibilitätsgen für SLE identifiziert. In einer Fall-Kontroll Studie wurde in dieser Arbeit untersucht, ob allele Varianten des CR2 eine erhöhte Suszeptibilität für SLE bewirken und ob die allelen Varianten eine Korrelation mit bestimmten klinischen Symptomen zeigen. In drei unabhängigen Populationen von SLE Patienten und einer entsprechenden Anzahl gesunder Kontrollpersonen konnten keine Hinweise für eine Assoziation zwischen den definierten allelen Varianten des CR2 und SLE nachgewiesen werden. Es konnte jedoch gezeigt werden, dass eine Korrelation zwischen den selteneren allelen Varianten des CR2 und einem späteren Zeitpunkt der Erstdiagnose von SLE besteht. Weiterhin gibt es Hinweise darauf, dass diese Variante mit einer verringerten Häufigkeit von Nierenbeteiligung bei SLE Patienten korreliert. Bei einer Analyse von humanen prä-B Zellen, die mit beiden allelen Hauptvatianten des Komplementrezeptors 2 transduziert wurden, konnte gezeigt werden, dass die seltenere Variante nicht wie erwartet eine negative Regulation der CD19 Oberflächendichte bewirkt. Dieses Ergebnis konnte durch eine Analyse der Oberflächendichte von CD19 und CD21 auf peripheren B Zellen von gesunden Normalspendern bestätigt werden. Mittels einer durchflusszytometrischen Untersuchung des Kalziumstroms in CD21 transduzierten prä-B Zellen nach Kreuzvernetzung des prä-BCR und simultaner Kreuzvernetzung des Komplementrezeptors 2 konnte gezeigt werden, dass die allelen Hauptvarianten den prä-BCR induzierten Kalziumstrom in unterschiedlicher Weise modulieren. Der Komplementrezeptor 2 stellt somit einen Modifikator für die Pathogenese des humanen SLE dar, dessen allele Varianten einen Einfluss auf das Alter bei SLE Erstdiagnose und das relative Verhältnis von CD19 zu CD21 auf der Oberfläche peripherer B Zellen haben, sowie ein verändertes Signalverhalten in prä-B Zellen mit ektop exprimiertem CD21 bewirken

NAD(P)H:quinone oxidoreductase 1 (NQO1) P187S polymorphism and prostate cancer risk in Caucasians

NAD(P)H:quinone oxidoreductase 1 (NQO1) catalyses the reduction of quinoid compounds to hydroquinones, preventing the generation of free radicals and reactive oxygen. A “C” to “T” transversion at position 609 of NQO1, leading to a nonsynonymous amino acid change (Pro187Ser, P187S), results in an altered enzyme activity. No NQO1 protein activity was detected in NQO1 609TT genotype, and low to intermediate activity was detected in NQO1 609CT genotype compared with 609CC genotype. Thus, this polymorphism may result in altered cancer predisposition. For prostate cancer, only sparse data are available. We therefore analyzed the distribution of the NQO1 P187S SNP (single nucleotide polymorphism) in prostate cancer patients and a healthy control group. Allelic variants were determined using RFLP analysis. Overall, 232 patients without any malignancy and 119 consecutive prostate cancer patients were investigated. The genotype distribution in our cohorts followed the Hardy–Weinberg equilibrium in cases and controls. The distribution of the NQO1 codon 187 SNP did not differ significantly between prostate cancer patients and the control group (p = 0.242). There was also no association between the allelic variants and stage or Gleason score of the tumors. The NQO1 P187S SNP was not significantly associated with an increased prostate cancer risk in our cohorts. The SNP has also no influence on histopathological characteristics of the tumors. A combined analysis of all available data from published European studies also showed no significant differences in the genotype distribution between controls and prostate cancer patients. Our data suggest a minor role of the NQO1 nucleotide 609 polymorphism in prostate carcinogenesis

Loss of RBMS1 as a regulatory target of miR-106b influences cell growth, gap closing and colony forming in prostate carcinoma

Abstract Prostate carcinoma (PCa) is the second most commonly diagnosed cancer in males worldwide. Among hereditary genetic mutations and nutrient factors, a link between the deregulation of microRNA (miRNA) expression and the development of prostate carcinoma is assumed. MiRNAs are small non-coding RNAs which post-transcriptionally regulate gene expression and which are involved in tumour development and progression as oncogenes or tumour suppressors. Although many genes could be confirmed as targets for deregulated miRNAs, the impact of differentially expressed miRNA and their regulatory target genes on prostate tumour development and progression are not fully understood yet. We could validate RBMS1, a barely described RNA-binding protein, as a new target gene for oncogenic miR-106b, which was identified as an induced miRNA in PCa. Further analysis revealed a loss of RBMS1 expression in prostate tumours compared to corresponding normal tissue. Overexpression of RBMS1 in DU145 and LNCaP prostate cancer cells resulted in diminished cell proliferation, colony forming ability as well as in retarded gap closing. Our results demonstrate for the first time a miR-106b dependent downregulation of RBMS1 in prostate carcinoma. Additionally, we show new tumour suppressive properties of RBMS1 whose observed loss may further elucidate the development of PCa

Expression of GP88 (Progranulin) Protein Is an Independent Prognostic Factor in Prostate Cancer Patients

Prostate cancer, the second most common cancer, is still a major cause of morbidity and mortality among men worldwide. The expression of the survival and proliferation factor progranulin (GP88) has not yet been comprehensively studied in PCa tumors. The aim of this study was to characterize GP88 protein expression in PCa by immunohistochemistry and to correlate the findings to the clinico-pathological data and prognosis. Immunohistochemical staining for GP88 was performed by TMA with samples from 442 PCa patients using an immunoreactive score (IRS). Altogether, 233 cases (52.7%) with negative GP88 staining (IRS < 2) and 209 cases (47.3%) with positive GP88 staining (IRS ≥ 2) were analyzed. A significant positive correlation was found for the GP88 IRS with the PSA value at prostatectomy and the cytoplasmic cytokeratin 20 IRS, whereas it was negatively associated with follow-up times. The association of GP88 staining with prognosis was further studied by survival analyses (Kaplan–Meier, univariate and multivariate Cox’s regression analysis). Increased GP88 protein expression appeared as an independent prognostic factor for overall, disease-specific and relapse-free survival in all PCa patients. Interestingly, in the subgroup of younger PCa patients (≤65 years), GP88 positivity was associated with a 3.8-fold (p = 0.004), a 6.0-fold (p = 0.008) and a 3.7-fold (p = 0.003) increased risk for death, disease-specific death and occurrence of a relapse, respectively. In the PCa subgroup with negative CK20 staining, GP88 positivity was associated with a 1.8-fold (p = 0.018) and a 2.8-fold increased risk for death and disease-specific death (p = 0.028). Altogether, GP88 protein positivity appears to be an independent prognostic factor for PCa patients

RNA Sequencing of Collecting Duct Renal Cell Carcinoma Suggests an Interaction between miRNA and Target Genes and a Predominance of Deregulated Solute Carrier Genes

Collecting duct carcinoma (CDC) is a rare renal cell carcinoma subtype with a very poor prognosis. There have been only a few studies on gene expression analysis in CDCs. We compared the gene expression profiles of two CDC cases with those of eight normal tissues of renal cell carcinoma patients. At a threshold of |log2fold-change| ≥1, 3349 genes were upregulated and 1947 genes were downregulated in CDCs compared to the normal samples. Pathway analysis of the deregulated genes revealed that cancer pathways and cell cycle pathways were most prominent in CDCs. The most upregulated gene was keratin 17, and the most downregulated gene was cubilin. Among the most downregulated genes were four solute carrier genes (SLC3A1, SLC9A3, SLC26A7, and SLC47A1). The strongest negative correlations between miRNAs and mRNAs were found between the downregulated miR-374b-5p and its upregulated target genes HIST1H3B, HK2, and SLC7A11 and between upregulated miR-26b-5p and its downregulated target genes PPARGC1A, ALDH6A1, and MARC2. An upregulation of HK2 and a downregulation of PPARGC1A, ALDH6A1, and MARC2 were observed at the protein level. Survival analysis of the cancer genome atlas (TCGA) dataset showed for the first time that low gene expression of MARC2, cubilin, and SLC47A1 and high gene expression of KRT17 are associated with poor overall survival in clear cell renal cell carcinoma patients. Altogether, we identified dysregulated protein-coding genes, potential miRNA-target interactions, and prognostic markers that could be associated with CDC

The Human Leukocyte Antigen G as an Immune Escape Mechanism and Novel Therapeutic Target in Urological Tumors

The non-classical human leukocyte antigen G (HLA-G) is a potent regulatory protein involved in the induction of immunological tolerance. This is based on the binding of membrane-bound as well as soluble HLA-G to inhibitory receptors expressed on various immune effector cells, in particular NK cells and T cells, leading to their attenuated functions. Despite its restricted expression on immune-privileged tissues under physiological conditions, HLA-G expression has been frequently detected in solid and hematopoietic malignancies including urological cancers, such as renal cell and urothelial bladder carcinoma and has been associated with progression of urological cancers and poor outcome of patients: HLA-G expression protects tumor cells from anti-tumor immunity upon interaction with its inhibitory receptors by modulating both the phenotype and function of immune cells leading to immune evasion. This review will discuss the expression, regulation, functional and clinical relevance of HLA-G expression in urological tumors as well as its use as a putative biomarker and/or potential therapeutic target for the treatment of renal cell carcinoma as well as urothelial bladder cancer

CCL2 Expression in Tumor Cells and Tumor-Infiltrating Immune Cells Shows Divergent Prognostic Potential for Bladder Cancer Patients Depending on Lymph Node Stage

Bladder cancer (BCa) is the ninth most commonly diagnosed cancer worldwide. Although there are several well-established molecular and immunological classifications, markers for tumor cells and immune cells that are associated with prognosis are still needed. The chemokine CC motif ligand 2 (CCL2) could be such a marker. We analyzed the expression of CCL2 by immunohistochemistry (IHC) in 168 muscle invasive BCa samples using a tissue microarray. Application of a single cut-off for the staining status of tumor cells (TCs; positive vs. negative) and immune cells (ICs; ≤6% of ICs vs. >6% of ICs) revealed 57 cases (33.9%) and 70 cases (41.7%) with CCL2-positive TCs or ICs, respectively. IHC results were correlated with clinicopathological and survival data. Positive CCL2 staining in TCs was associated with shorter overall survival (OS), disease-specific survival (DSS), and relapse-free survival (RFS) (p = 0.004, p = 0.036, and p = 0.047; log rank test) and appeared to be an independent prognostic factor for OS (RR = 1.70; p = 0.007; multivariate Cox’s regression analysis). In contrast, positive CCL2 staining in the ICs was associated with longer OS, DSS, and RFS (p = 0.032, p = 0.001, and p = 0.001; log rank test) and appeared to be an independent prognostic factor for DSS (RR = 1.77; p = 0.031; multivariate Cox’s regression analysis). Most interestingly, after separating the patients according to their lymph node status (N0 vs. N1+2), CCL2 staining in the ICs was differentially associated with prognosis. In the N0 group, CCL2 positivity in the ICs was a positive independent prognostic factor for OS (RR = 1.99; p = 0.014), DSS (RR = 3.17; p = 0.002), and RFS (RR = 3.10; p = 0.002), whereas in the N1+2 group, CCL2 positivity was a negative independent factor for OS (RR = 3.44; p = 0.019)) and RFS (RR = 4.47; p = 0.010; all multivariate Cox’s regression analyses). In summary, CCL2 positivity in TCs is a negative prognostic factor for OS, and CCL2 can mark ICs that are differentially associated with prognosis depending on the nodal stage of BCa patients. Therefore, CCL2 staining of TCs and ICs is suggested as a prognostic biomarker for BCa patients

Clean Intermittent Catheterization in Children under 12 Years Does Not Have a Negative Impact on Long-Term Graft Survival following Pediatric Kidney Transplantation

Background: Congenital anomalies of the kidneys and urinary tract (CAKUTs) are one of the most prevalent primary causes of end-stage renal disease (ESRD) in young children, and approximately one-third of these children present with lower urinary tract dysfunction (LUTD). Many children with LUTD require therapy with clean intermittent catheterization (CIC). CIC commonly leads to bacteriuria, and considerations have arisen regarding whether CIC in immunosuppressed children is safe or whether repeated febrile urinary tract infections (UTIs) may lead to the deterioration of kidney graft function. Material and Methods: We retrospectively reviewed all cases of primary kidney transplantation performed in our center between 2001 and 2020 in recipients aged less than twelve years. The number of episodes of febrile UTIs as well as the long-term kidney graft survival of children undergoing CIC were compared to those of children with urological causes of ESRD not undergoing CIC, as well as to those of children with nonurological causes of ESRD. Results: Following successful kidney transplantation in 41 children, CIC was needed in 8 of these patients. These 8 children undergoing CIC had significantly more episodes of febrile UTIs than did the 18 children with a nonurological cause of ESRD (p = 0.04) but not the 15 children with a urological cause of ESRD who did not need to undergo CIC (p = 0.19). Despite being associated with a higher rate of febrile UTIs, CIC was not identified as a risk factor for long-term kidney graft survival, and long-term graft survival did not significantly differ between the three groups at a median follow-up of 124 months. Conclusions: Our study demonstrates that, under regular medical care, CIC following pediatric transplantation is safe and is not associated with a higher rate of long-term graft loss.</jats:p

Expression of human Piwi-like genes is associated with prognosis for soft tissue sarcoma patients

Background Argonaute genes are essential for RNA interference, stem cell maintenance and differentiation. The Piwi-like genes, a subclass of the Argonaute genes, are expressed mainly in the germline. These genes may be re-expressed in tumors, and expression of the Piwi-like genes is associated with prognosis in several types of tumors. Methods We measured the expression of Piwi-like mRNAs (Piwi-like 2–4) in 125 soft tissue sarcoma (STS) samples by qPCRs. Statistical tests were applied to study the correlation of expression levels with tumor-specific survival for STS patients. Results In multivariate Cox’s regression analyses, we showed that low Piwi-like 2 and Piwi-like 4 mRNA expression were significantly associated with a worse prognosis (RR = 1.87; p = 0.032 and RR = 1.82; p = 0.039). Low expression of both genes was associated with a 2.58-fold increased risk of tumor-related death (p = 0.01). Piwi-like 4 and combined Piwi-like 2 and 4 mRNA levels correlated significantly with prognosis (RR = 3.53; p = 0.002 and RR = 5.23; p = 0.004) only for female but not for male patients. However, combined low Piwi-like 2 and 3 transcript levels were associated with worse survival (RR = 5.90; p = 0.02) for male patients. Conclusions In this study, we identified a significant association between the expression of Piwi-like 2 and 4 mRNAs and the tumor-specific survival of soft tissue sarcoma patients. Furthermore, a connection between sex and the impact of Piwi-like mRNA expressions on STS patients’ prognosis was shown for the first time

Prognostic impact of molecular muscle-invasive bladder cancer subtyping approaches and correlations with variant histology in a population-based mono-institutional cystectomy cohort

Purpose Recently discovered molecular classifications for urothelial bladder cancer appeared to be promising prognostic and predictive biomarkers. The present study was conducted to evaluate the prognostic impact of molecular subtypes assessed by two different methodologies (gene and protein expression), to compare these two approaches and to correlate molecular with histological subtypes in a consecutively collected, mono-institutional muscle-invasive bladder cancer (MIBC) cohort. Methods 193 MIBC were pathologically re-evaluated and molecular subtypes were assessed on mRNA (NanoString technology, modified 21-gene-containing MDACC approach) and protein levels (immuno-histochemical [IHC] analysis of CK5, CK14, CD44, CK20, GATA3 and FOXA1). Descriptive statistical methods and uni-/multi-variable survival models were employed to analyze derived data. Results Neither gene expression nor protein-based subtyping showed significant associations with disease-specific (DSS) or recurrence-free survival (RFS). Agreement between mRNA (reference) and protein-based subtyping amounted 68.6% for basal, 76.1% for luminal and 50.0% for double-negative tumors. Histological subtypes associated with RFS in uni-variable (P = 0.03), but not in multivariable survival analyses. Tumors with variant histology predominantly showed luminal subtypes (gene expression subtyping: 36/55 cases, 65.5%; protein subtyping: 44/55 cases, 80.0%). Squamous differentiation significantly associated with basal subtypes (gene expression subtyping: 44/45 squamous cases, 97.8%; protein subtyping: 36/45 cases, 80.0%). Conclusion In our consecutive cystectomy cohort, neither gene, protein expression-based subtyping, nor histological subtypes associated with DSS or RFS in multi-variably adjusted survival analyses. Application of a limited IHC subtyping marker panel showed high concordance of 83.9% with gene expression-based subtyping, thus underlining the utility for subtyping in pathological routine diagnostics. In addition, histological MIBC subtypes are strong indicators for intrinsic subtypes