Direct conversion of mouse embryonic fibroblasts into functional keratinocytes through transient expression of pluripotency-related genes

The insufficient ability of specialized cells such as neurons, cardiac myocytes, and epidermal cells to regenerate after tissue damage poses a great challenge to treat devastating injuries and ailments. Recent studies demonstrated that a diverse array of cell types can be directly derived from embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs), or somatic cells by combinations of specific factors. The use of iPSCs and direct somatic cell fate conversion, or transdifferentiation, holds great promise for regenerative medicine as these techniques may circumvent obstacles related to immunological rejection and ethical considerations. However, producing iPSC-derived keratinocytes requires a lengthy two-step process of initially generating iPSCs and subsequently differentiating into skin cells, thereby elevating the risk of cellular damage accumulation and tumor formation. In this study, we describe the reprogramming of mouse embryonic fibroblasts into functional keratinocytes via the transient expression of pluripotency factors coupled with directed differentiation. The isolation of an iPSC intermediate is dispensable when using this method. Cells derived with this approach, termed induced keratinocytes (iKCs), morphologically resemble primary keratinocytes. Furthermore they express keratinocyte-specific markers, downregulate mesenchymal markers as well as the pluripotency factors Oct4, Sox2, and Klf4, and they show important functional characteristics of primary keratinocytes. iKCs can be further differentiated by high calcium administration in vitro and are capable of regenerating a fully stratified epidermis in vivo. Efficient conversion of somatic cells into keratinocytes could have important implications for studying genetic skin diseases and designing regenerative therapies to ameliorate devastating skin conditions.COST (European Cooperation in Science and Technology) (EU-COST Action BM1302 “Joining Forces in Corneal Regeneration Research”)University of Cypru

VC-backed Biotechnology Firms: What is Entrepreneurs’ Return?

Valuation is a highly significant process used extensively not only by start-ups seeking Venture Capital (VC) funding but also by large, established companies that are in need to set a price for an acquisition or merger. Such process becomes increasingly complex in the biotechnology sector where the realization of the true value of a firm is largely correlated with the success or failure of lead drug candidates undergoing clinical trials. In this article, the author sets up a simple long-term financial model that aims at providing an easy tool for estimating entrepreneurs’ return at exit.</jats:p

Abstract B47: Biomarker discovery and evaluation of response to anticancer therapeutics in breast cancer using a novel nanofluidic immunoassay

Abstract Cancer subtype specific molecular variations dramatically affect patient responses to already existing treatments. For example, the phosphorylation status of many proteins that are involved in signal transduction pathways perturbed in cancer cells is extremely important in determining whether these cells are susceptible to killing by available cancer therapeutics. Therefore, differentially phosphorylated protein isoforms can be a particularly useful prognostic biomarker of drug response in the clinic. However, accurate detection and quantitative analysis of cancer-related phosphoproteins in tumors is limited by current technologies. Using a novel, fully automated nanocapillary electrophoresis technology (CB1000TM) designed to separate protein molecules based on their isoelectric point, we are currently developing highly sensitive assays for reliable assessment of the phosphorylation status of cancer-related phosphoproteins in tumors, before and during drug treatment. We have already developed and optimized assays measuring all AKT, ERK and MEK isoforms and their respective phosphoisoforms using a single detection reagent method. Using these assays, we were able to measure levels of activated MEK1/2, ERK1/2 and AKT1/2/3 in cancer cell lines, mouse xenografts and patient FNAs using protein extracted from as few as 125 cells. We have already measured and characterized changes in the phosphorylation states of these targets before, during and after treatment with inhibitors of the PI3K pathway in breast cancer, and based on this data we are currently developing molecular profiles that predict response to available therapeutics. Citation Information: Clin Cancer Res 2010;16(7 Suppl):B47</jats:p

Whole transcriptome sequence data of 5-FU sensitive and 5-FU resistant tumors generated in a mouse model of de novo carcinogenesis

We have performed whole transcriptome sequencing of 5-FU resistant and 5-FU sensitive tumors generated in a mouse model of de novo carcinogenesis that closely recapitulates tumor initiation, progression and maintenance in vivo. Tumors were generated using the DMBA/TPA model of chemically induced carcinogenesis [1], tumor-bearing mice were subsequently treated with 5-FU, and tumor growth as well as response to treatment was monitored by measuring tumor volume twice a week. Based on these measurements, we selected two 5-FU resistant and two 5-FU sensitive tumors and performed whole transcriptome sequencing and in order to identify differentially expressed transcripts between the two sets. Data obtained is deposited and available through NCBI SRA (reference number SRP155180 – https://www.ncbi.nlm.nih.gov/sra/?term=SRP155180)

Shared mechanisms in stemness and carcinogenesis: lessons from oncogenic viruses

A rise in technologies for epigenetic reprogramming of cells to pluripotency, highlights the potential of understanding and manipulating cellular plasticity in unprecedented ways. Increasing evidence points to shared mechanisms between cellular reprogramming and the carcinogenic process, with the emerging possibility to harness these parallels in future therapeutics. In this review, we present a synopsis of recent work from oncogenic viruses which contributes to this body of knowledge, establishing a nexus between infection, cancer, and stemness

Critical Role for Arginine Methylation in Adenovirus-Infected Cells▿

During the late stages of adenovirus infection, the 100K protein (100K) inhibits the translation of cellular messages in the cytoplasm and regulates hexon trimerization and assembly in the nucleus. However, it is not known how it switches between these two functions. Here we show that 100K is methylated on arginine residues at its C terminus during infection and that this region is necessary for binding PRMT1 methylase. Methylated 100K is exclusively nuclear. Mutation of the third RGG motif (amino acids 741 to 743) prevents localization to the nucleus during infection, suggesting that methylation of that sequence is important for 100K shuttling. Treatment of infected cells with methylation inhibitors inhibits expression of late structural proteins. These data suggest that arginine methylation of 100K is necessary for its localization to the nucleus and is a critical cellular function necessary for productive adenovirus infection

A critical examination of the principal theories of the Hebrew verbal system between 1827 and 1954 with particular reference to the waw consecutive problem

SIGLEAvailable from British Library Document Supply Centre- DSC:D42136/82 / BLDSC - British Library Document Supply CentreGBUnited Kingdo