Major clinical issues in bladder cancer include the identification of prediction markers and novel therapeutic targets for invasive bladder cancer. In the current study, we describe the isolation and characterization of a tumor-initiating cell (T-IC) subpopulation in primary human bladder cancer, based on the expression of markers similar to that of normal bladder basal cells (Lineage-CD44 ؉ CK5 ؉ CK20 ؊ ). The bladder T-IC subpopulation was defined functionally by its enriched ability to induce xenograft tumors in vivo that recapitulated the heterogeneity of the original tumor. Further, molecular analysis of more than 300 bladder cancer specimens revealed heterogeneity among activated oncogenic pathways in T-IC (e.g., 80% Gli1, 45% Stat3, 10% Bmi-1, and 5% ␤-catenin). Despite this molecular heterogeneity, we identified a unique bladder T-IC gene signature by gene chip analysis. This T-IC gene signature, which effectively distinguishes muscle-invasive bladder cancer with worse clinical prognosis from non-muscle-invasive (superficial) cancer, has significant clinical value. It also can predict the progression of a subset of recurring non-muscleinvasive cancers. Finally, we found that CD47, a protein that provides an inhibitory signal for macrophage phagocytosis, is highly expressed in bladder T-ICs compared with the rest of the tumor. Blockade of CD47 by a mAb resulted in macrophage engulfment of bladder cancer cells in vitro. In summary, we have identified a T-IC subpopulation with potential prognostic and therapeutic value for invasive bladder cancer.antibody therapy ͉ bladder cancer ͉ cancer stem cell ͉ CD44 ͉ CD47

Type I (α, β) and type II (γ) interferons (IFN) possess pleiotropic functions, including antiviral, antiproliferative and immunomodulatory activities, which are likely mediated by the induction of unique subsets of IFN-stimulated genes (ISG) that are differentially regulated by IFN subtypes and in different cell types. Oligonucleotide arrays (Affymetrix) were used to investigate differential gene expression in the human fibrosarcoma cell line, HT1080, or primary mouse embryonic fibroblasts in response to IFNs or virus infection. In all samples, the detection of known IFN-regulated genes helped to validate the experimental design and conditions. Combined analysis of these data sets, each with several hundreds of differentially expressed genes, has led to the idenfication of groups of IFN-regulated genes with similar functions, previously unknown IFN-regulated pathways and potential to crosstalk with other signalling pathways. For example, ISGs with similar functions included apoptosis regulators, chemokines, genes involved with antigen processing and presentation, GTPases, heat shock proteins and members of the ubiquitin system. Signalling molecules identified as potentially novel ISGs included known kinases, phosphatases and signalling adapter molecules. The functional significance of each gene relating to IFN functions requires further study on an individual basis, but the continuous refinement of a global view for IFN-regulated pathways may identify additional yet unknown areas of IFN action and enable better design of therapeutic strategies based on IFN-regulated mechanisms.We describe a rapid and efficient method for identifying secreted and transmembrane gene products. From yeast and human cell cultures, mRNA species bound to rough endoplasmic reticulum (rER)-associated polysomes were separated from free and cytoplasmic polysome-associated mRNAs by sedimentation equilibrium or sedimentation velocity. The representation of mRNA species in each population was quantified by hybridization to high-density DNA microarrays. Transcripts known to encode transmembrane or secreted proteins were significantly enriched in the rER-associated fractions, whereas those known to encode cytoplasmic proteins were enriched in the fractions containing free and cytoplasmic polysome-associated mRNAs. Based on our data, we are able to predict with reasonable confidence that over 275 human and 300 yeast genes encode previously unrecognized transmembrane, ER/golgi-resident or secreted proteins. The combination of fractionation of nucleic acids on the basis of physical properties and analysis on DNA microarrays is a general method that can be applied to a diverse set of biological questions.A basic problem in multivariate systems is finding predictive relations between the measurable variables in the system. The extent to which a target variable can be predicted via measurement of a set of predictor variables reveals the level of interaction between the variables. We want to quantify the degree to which the target variable is statistic...

This Review Article provides a multi-stakeholder view on the current status of neoadjuvant therapy in lung cancer. Given the success of oncogene-targeted therapy and immunotherapy for patients with advanced lung cancer, there is a renewed interest in studying these agents in earlier disease settings with the opportunity to have an even greater impact on patient outcomes. There are unique opportunities and challenges with the neoadjuvant approach to drug development. To achieve more rapid knowledge turns, study designs, endpoints, and definitions of pathologic response should be standardized and harmonized. Continued dialogue with all stakeholders will be critical to design and test novel induction strategies, which could expedite drug development for patients with early lung cancer who are at high risk for metastatic recurrence. Published by Elsevier Inc. on behalf of International Association for the Study of Lung Cancer.

Innovation and progress in radiation oncology depend on discovery and insights realized through research in radiation biology. Radiobiology research has led to fundamental scientific insights, from the discovery of stem/progenitor cells to the definition of signal transduction pathways activated by ionizing radiation that are now recognized as integral to the DNA damage response (DDR). Radiobiological discoveries are guiding clinical trials that test radiation therapy combined with inhibitors of the DDR kinases DNA-dependent protein kinase (DNA-PK), ataxia telangiectasia mutated (ATM), ataxia telangiectasia related (ATR), and immune or cell cycle checkpoint inhibitors. To maintain scientific and clinical relevance, the field of radiation biology must overcome challenges in research workforce, training, and funding. The National Cancer Institute convened a workshop to discuss the role of radiobiology research and radiation biologists in the future scientific enterprise. Here, we review the discussions of current radiation oncology research approaches and areas of scientific focus considered important for rapid progress in radiation sciences and the continued contribution of radiobiology to radiation oncology and the broader biomedical research community.

At central synapses, P͞Q-type Ca 2؉ channels normally provide a critical Ca 2؉ entry pathway for neurotransmission. Nevertheless, we found that nerve terminals lacking ␣1A (CaV2.1), the pore-forming subunit of P͞Q-type channels, displayed a remarkable preservation of synaptic function. Two consistent physiological changes reflective of synaptic homeostasis were observed in cultured hippocampal neurons derived from ␣1A (؊/؊) mice. First, the presynaptic response to an ionophore-mediated Ca 2؉ elevation was 50% greater, indicating an enhanced Ca 2؉ sensitivity of the release machinery. Second, basal miniature excitatory postsynaptic current frequency in ␣1A (؊/؊) neurons was increased 2-fold compared with WT neurons and occluded the normal response of presynaptic terminals to cAMP elevation, suggesting that the compensatory mechanism in ␣1A (؊/؊) synapses and the modulation of presynaptic function by PKA might share a final common pathway. We used cDNA microarray analysis to identify molecular changes underlying homeostatic regulation in the ␣1A (؊/؊) hippocampus. The 40,000 entries in our custom-made array included likely targets of presynaptic homeostasis, along with many other transcripts, allowing a wide-ranging examination of gene expression. The developmental pattern of changes in transcript levels relative to WT was striking; mRNAs at 5 and 11 days postnatal showed little deviation, but clear differences emerged by 22 days. Many of the transcripts that differed significantly in abundance corresponded to known genes that could be incorporated within a logical pattern consistent with the modulation of presynaptic function. Changes in endocytotic proteins, signal transduction kinases, and candidates for Ca 2؉ -sensing molecules were consistent with implications of the direct physiological experiments.