Opinion of German Immunologists on SARS-CoV-2: Results of an Online Survey.

Background Little is known about the opinion of professional academic immunologists regarding the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic. Methodology In this study, we designed an online survey to determine the opinion of immunologically competent academics on SARS-CoV-2 compared with seasonal flu (the infection fatality rate, infectivity, the challenge to the health system, the importance of vaccine development, and the importance of the virulence of the virus and host factors), in addition to collecting demographic status variables and information sources used. Links to the survey were sent to all German-speaking immunologists, bacteriologists, virologists, and infectiologists in Germany, Austria, and Switzerland. Results A total of 91 full datasets were returned after three waves of requests. Approximately half of the respondents were male and half were more junior. Slightly more than half of the respondents said that the infection fatality rate and the infectivity were higher compared to flu, and 82% said that the challenge to the health system is higher. Overall, 52% found that the immune system is more important than the virus, and a majority (59%) supported the current practice of vaccination development by telescoping. A majority were of the view that conspiracy theories and non-pharmacological interventions pose a greater danger than the virus. Respondents who were more junior but well-published and mostly informed by public channels were more likely to support a mainstream view. Conclusions German-speaking immunological professionals hold widely diverging opinions regarding SARS-CoV-2. Over half of the surveyed professionals considered SARS-CoV-2 to be more dangerous and infective than the seasonal flu. However, the majority considered the health system to be under higher strain. Interestingly, more than half of them found host factors more important.

A Cell-Based Method for Identification of Chemotherapy Resistance Cancer Genes.

Here we describe a method for identifying genes and genetic pathways responsible for chemoresistance in cancer cells. The method is based on generation and characterization of matched pairs of chemotherapy-sensitive/chemotherapy-resistant cancer cell lines. In this protocol we are using endometrial cancer cell lines treated with carboplatin and paclitaxel, which are first-line chemotherapies for gynecologic malignancies. The chemoresistant cells and their chemosensitive counterparts are used for downstream applications including bulk RNA-sequencing analysis to identify a set of genes and pathways that are associated with chemoresistance. Identification of pathways responsible for innate or acquired chemoresistance is of paramount importance for the identification of biomarkers for cancer risk stratification and prognosis, and as a pharmacogenomics model for identification of alternative chemotherapy approaches for treatment of patients with recurrent and chemoresistant disease.

RNA Sequencing of Carboplatin- and Paclitaxel-Resistant Endometrial Cancer Cells Reveals New Stratification Markers and Molecular Targets for Cancer Treatment.

Despite advances in surgical technique and adjuvant treatment, endometrial cancer has recently seen an increase in incidence and mortality in the USA. The majority of endometrial cancers can be cured by surgery alone or in combination with adjuvant chemo- or radiotherapy; however, a subset of patients experience recurrence for reasons that remain unclear. Recurrence is associated with chemoresistance to carboplatin and paclitaxel and consequentially, high mortality. Understanding the pathways involved in endometrial cancer chemoresistance is paramount for the identification of biomarkers and novel molecular targets for this disease. Here, we generated the first matched pairs of carboplatin-sensitive/carboplatin-resistant and paclitaxel-sensitive/paclitaxel-resistant endometrial cancer cells and subjected them to bulk RNA sequencing analysis. We found that 45 genes are commonly upregulated in carboplatin- and paclitaxel-resistant cells as compared to controls. Of these, the leukemia inhibitory factor, (LIF), the protein tyrosine phosphatase type IVA, member 3 (PTP4A3), and the transforming growth factor beta 1 (TGFB1) showed a highly significant correlation between expression level and endometrial cancer overall survival (OS) and can stratify the 545 endometrial cancer patients in the TCGA cohort into a high-risk and low-risk-cohorts. Additionally, four genes within the 45 upregulated chemoresistance-associated genes are ADAMTS5, MICAL2, STAT5A, and PTP4A3 codes for proteins for which small-molecule inhibitors already exist. We identified these proteins as molecular targets for chemoresistant endometrial cancer and showed that treatment with their correspondent inhibitors effectively killed otherwise chemoresistant cells. Collectively, these findings underline the utility of matched pair of chemosensitive and chemoresistant cancer cells to identify markers for endometrial cancer risk stratification and to serve as a pharmacogenomics model for identification of alternative chemotherapy approaches for treatment of patients with recurrent disease.

UNC-45A is required for neurite extension via controlling NMII activation.

UNC-45A is a highly conserved member of the UNC-45/CRO1/She4p family of proteins, which act as chaperones for conventional and nonconventional myosins. NMII mediates contractility and actin-based motility, which are fundamental for proper growth cone motility and neurite extension. The presence and role of UNC-45A in neuronal differentiation have been largely unknown. Here we demonstrate that UNC-45A is a novel growth cone--localized, NMII-associated component of the multiprotein complex regulating growth cone dynamics. We show that UNC-45A is dispensable for neuron survival but required for neurite elongation. Mechanistically, loss of UNC-45A results in increased levels of NMII activation. Collectively our results provide novel insights into the molecular mechanisms of neurite growth and define UNC-45A as a novel and master regulator of NMII-mediated cellular processes in neurons.

Single cell sequencing reveals heterogeneity within ovarian cancer epithelium and cancer associated stromal cells.

OBJECTIVES: The purpose of this study was to determine the level of heterogeneity in high grade serous ovarian cancer (HGSOC) by analyzing RNA expression in single epithelial and cancer associated stromal cells. In addition, we explored the possibility of identifying subgroups based on pathway activation and pre-defined signatures from cancer stem cells and chemo-resistant cells. METHODS: A fresh, HGSOC tumor specimen derived from ovary was enzymatically digested and depleted of immune infiltrating cells. RNA sequencing was performed on 92 single cells and 66 of these single cell datasets passed quality control checks. Sequences were analyzed using multiple bioinformatics tools, including clustering, principle components analysis, and geneset enrichment analysis to identify subgroups and activated pathways. Immunohistochemistry for ovarian cancer, stem cell and stromal markers was performed on adjacent tumor sections. RESULTS: Analysis of the gene expression patterns identified two major subsets of cells characterized by epithelial and stromal gene expression patterns. The epithelial group was characterized by proliferative genes including genes associated with oxidative phosphorylation and MYC activity, while the stromal group was characterized by increased expression of extracellular matrix (ECM) genes and genes associated with epithelial-to-mesenchymal transition (EMT). Neither group expressed a signature correlating with published chemo-resistant gene signatures, but many cells, predominantly in the stromal subgroup, expressed markers associated with cancer stem cells. CONCLUSIONS: Single cell sequencing provides a means of identifying subpopulations of cancer cells within a single patient. Single cell sequence analysis may prove to be critical for understanding the etiology, progression and drug resistance in ovarian cancer.