Amnah Othman

A Comprehensive Analysis of Human Gene Expression Profiles Identifies Stromal Immunoglobulin κ C as a Compatible Prognostic Marker in Human Solid Tumors

Purpose: Although the central role of the immune system for tumor prognosis is generally accepted, a single robust marker is not yet available. Experimental Design: On the basis of receiver operating characteristic analyses, robust markers were identified from a 60-gene B cell–derived metagene and analyzed in gene expression profiles of 1,810 breast cancer; 1,056 non–small cell lung carcinoma (NSCLC); 513 colorectal; and 426 ovarian cancer patients. Protein and RNA levels were examined in paraffin-embedded tissue of 330 breast cancer patients. The cell types were identified with immunohistochemical costaining and confocal fluorescence microscopy. Results: We identified immunoglobulin κ C (IGKC) which as a single marker is similarly predictive and prognostic as the entire B-cell metagene. IGKC was consistently associated with metastasis-free survival across different molecular subtypes in node-negative breast cancer (n = 965) and predicted response to anthracycline-based neoadjuvant chemotherapy (n = 845; P < 0.001). In addition, IGKC gene expression was prognostic in NSCLC and colorectal cancer. No association was observed in ovarian cancer. IGKC protein expression was significantly associated with survival in paraffin-embedded tissues of 330 breast cancer patients. Tumor-infiltrating plasma cells were identified as the source of IGKC expression. Conclusion: Our findings provide IGKC as a novel diagnostic marker for risk stratification in human cancer and support concepts to exploit the humoral immune response for anticancer therapy. It could be validated in several independent cohorts and carried out similarly well in RNA from fresh frozen as well as from paraffin tissue and on protein level by immunostaining. Clin Cancer Res; 18(9); 2695–703. ©2012 AACR.

The prognostic relevance of tumour-infiltrating plasma cells and immunoglobulin kappa C indicates an important role of the humoral immune response in non-small cell lung cancer

A prognostic impact of immunoglobulin kappa C (IGKC) expression has been described in cancer. We analysed the influence of B-cell and plasma cell markers, as well as IGKC expression, in non-small lung cancer (NSCLC) using immunohistochemistry on a tissue microarray. IGKC protein expression was independently associated with longer survival, with particular impact in the adenocarcinoma subgroup. Moreover, a correlation was seen with CD138+ cells, but not with CD20. CD138 expression revealed a comparable association with survival. In conclusion, IGKC expression in stroma-infiltrating plasma cells is a prognostic marker in NSCLC, supporting emerging treatment concepts that exploit the humoral immune response.

Phenotype and growth behavior of residual β-catenin-positive hepatocytes in livers of β-catenin-deficient mice

Signaling through the Wnt/β-catenin pathway is a crucial determinant of hepatic zonal gene expression, liver development, regeneration, and tumorigenesis. Transgenic mice with hepatocyte-specific knockout of Ctnnb1 (encoding β-catenin) have proven their usefulness in elucidating these processes. We now found that a small number of hepatocytes escape the Cre-mediated gene knockout in that mouse model. The remaining β-catenin-positive hepatocytes showed approximately 25% higher cell volumes compared to the β-catenin-negative cells and exhibited a marker protein expression profile similar to that of normal perivenous hepatocytes or hepatoma cells with mutationally activated β-catenin. Surprisingly, the expression pattern was observed independent of the cell’s position within the liver lobule, suggesting a malfunction of physiological periportal repression of perivenously expressed genes in β-catenin-deficient liver. Clusters of β-catenin-expressing hepatocytes lacked expression of the gap junction proteins Connexin 26 and 32. Nonetheless, β-catenin-positive hepatocytes had no striking proliferative advantage, but started to grow out on treatment with phenobarbital, a tumor-promoting agent known to facilitate the formation of mouse liver adenoma with activating mutations of Ctnnb1. Progressive re-population of Ctnnb1 knockout livers with wild-type hepatocytes was seen in aged mice with a pre-cirrhotic phenotype. In these large clusters of β-catenin-expressing hepatocytes, perivenous-specific gene expression was re-established. In summary, our data demonstrate that the zone-specificity of a hepatocyte’s gene expression profile is dependent on the presence of β-catenin, and that β-catenin provides a proliferative advantage to hepatocytes when promoted with phenobarbital, or in a pre-cirrhotic environment.

Modulating Portal Hemodynamics With Vascular Ring Allows Efficient Regeneration After Partial Hepatectomy in a Porcine Model

Objective: To investigate safety and efficacy of temporary portal hemodynamics modulation with a novel percutaneously adjustable vascular ring (MID-AVR) onto a porcine model of 75% hepatectomy. Background: Postoperative liver failure is a leading cause of mortality after major hepatectomy. Portal flow modulation is an increasingly accepted concept to prevent postoperative liver failure. Nonetheless, the current strategies have shortcomings. Methods: Resection was performed under hemodynamic monitoring in 17 large, white pigs allocated into 2 groups. Eight pigs had ring around the portal vein for 3 days with the aim of reducing changes in hemodynamics due to hepatectomy. Analysis of hemodynamics, laboratory, and histopathological parameters was performed. Results: Percutaneous inflation, deflation, and removal of the MID-AVR were safe. Two (25%) pigs in the MID-AVR group and 4 (45%) controls died before day 3 (P = NS). A moderate increase of portal flow rate per liver mass after resection was associated with better survival (P = 0.017). The portocaval pressure gradient was lower after hepatectomy in the MID-AVR group (P = 0.001). Postoperative serum bilirubin levels were lower in the MID-AVR group (P = 0.007 at day 5). In the MID-AVR group, the Ki67 index was significantly higher on day 3 (P = 0.043) and the architectural derangement was lower (P < 0.05). Morphometric quantification of the bile canaliculi revealed a significantly lower number of intersection branches (P < 0.05) and intersection nodes (P < 0.001) on day 7 compared with the preoperative specimen, in the control group. These differences were not found in the ring group. Conclusions: MID-AVR is safe for portal hemodynamics modulation. It might improve liver regeneration by protecting liver microarchitecture.

Highlight report: hepatotoxicity prediction with Hep3B cells

Recently, Yu et al. (2018) have contributed a study with 3D-cultivated Hep3B cells tested with 22 model compounds. LC50 values were compared to rat LD50 values and human Cmax values. At a first glance, one may ask, whether it is really justified to introduce another liver tumor cell line besides the already well-established HepG2 and HepaRG cells. However, the obtained LC50 values were remarkably low. For example, acetaminophen resulted in an LC50 of 300 μM, which is lower compared to many previously reported cell systems, such as HepG2. Finally, a remarkably good overall predictively of human hepatotoxicity was obtained. Currently, human hepatocytes are still considered a gold standard for in vitro testing of human hepatotoxicity (Pfeiffer et al. 2015; Leist et al. 2017; Grinberg et al. 2014; Ghallab 2017; Hammad and Ahmed 2014; Hammad et al. 2015). Moreover, hepatotoxicity represents a complex process that may involve interaction of several cell types (Jansen et al. 2017; Vartak et al. 2016; Albrecht 2017; Schenk et al. 2017). If the high predictively of the Hep3Bbased test system (Yu et al. 2018) would be confirmed by further compounds, it might indeed facilitate prediction of hepatotoxicity. However, whether prediction of idiosyncratic hepatotoxicity will be possible based on these still remains to be shown. Compliance with ethical standards

From basic research to applied veterinary sciences: current status, challenges and perspectives

Globally, veterinary-based research is performed for two purposes. First, it is performed to better understand the molecular, genetic and phenotypic, as well as pathophysiological background of animal diseases. This discipline has a great value economically, i.e., controlling diseases (Porphyre et al. 2018), increasing animal productivity (Abdel-Wareth et al. 2014) and decreasing the required health care costs (Prinsloo et al. 2018). However, more importantly, clinically to prevent human diseases of animal origin, i.e., communicable/zoonotic diseases, food borne illness (Hassan et al. 2011), toxic residues (Mohamed et al. 2017), etc. Recently, several measures were considered to improve animal farms and health care (Jacobs et al. 2018). However, more attention should be paid to protect wild, farm and companion animals, as well as pets, from genetically modified microorganisms (Elhariry et al. 2011), toxic residues (Mohamed et al. 2017) and climate alterations (Combe et al. 2017). Moreover, vaccine development and approval of less resistible compounds, i.e., antibiotics and cancer chemotherapeutics, are the most important protective and preventative strategies (Kumar et al. 2018) and must be considered. Second, animals are intensively used as experimental models to understand the underlying mechanisms of human diseases (Nussler et al. 2014), to identify new targetable protein(s) or pathway(s) and to approve drugs preclinically (Henderson et al. 2013). In the last decades, numerous funding agencies, such as the EU, suggested that animals used in experiments must be replaced, reduced and refined—the so called 3Rs (Daneshian et al. 2015). Basically, there are two ways to achieve these ambitious goals: (1) to develop alternative wet lab-based methods, e.g., 2D cell culture (Godoy et al. 2016), 3D spheroid models (Kim et al. 2015), primary cells (Reif et al. 2015) and cell lines (Luckert et al. 2017), or (2) to use in silico computer-assisted models to answer the corresponding biological questions (Hammad et al. 2015). Despite extensive funding in the last decades for both methods, the optimal understanding of human pathophysiological events is still unmet. We do not believe that animals will be completely replaced by an alternative method; however, research is still leading in this direction every day. Based on the aforementioned background, animal science is a significantly rich field of research. In approximately 90% of biomedical oriented articles in the last decade, experimental animals, primary cells or cell lines of animal origin were involved. Therefore, the Faculty of Seddik Hammad’s contribution was exclusively achieved at the Faculty of Veterinary Medicine, South Valley University, Egypt.