[Letter to the Editor] The need for improved education and training in research antibody usage and validation practices.

Address correspondence to Leonard P. Freedman, Global Biological Standards Institute, 1020 19th Street, NW, Suite 550, Washington, DC, 20036. E-mail: lfreedman@gbsi.org.

Computational and bioinformatics frameworks for next-generation whole exome and genome sequencing.

It has become increasingly apparent that one of the major hurdles in the genomic age will be the bioinformatics challenges of next-generation sequencing. We provide an overview of a general framework of bioinformatics analysis. For each of the three stages of (1) alignment, (2) variant calling, and (3) filtering and annotation, we describe the analysis required and survey the different software packages that are used. Furthermore, we discuss possible future developments as data sources grow and highlight opportunities for new bioinformatics tools to be developed.

Tissue microarrays and quantitative tissue-based image analysis as a tool for oncology biomarker and diagnostic development.

INTRODUCTION: Translational oncology has been improved by using tissue microarrays (TMAs), which facilitate biomarker analysis of large cohorts on a single slide. This has allowed for rapid analysis and validation of potential biomarkers for prognostic and predictive value, as well as for evaluation of biomarker prevalence. Coupled with quantitative analysis of immunohistochemical (IHC) staining, objective and standardized biomarker data from tumor samples can further advance companion diagnostic approaches for the identification of drug-responsive or resistant patient subpopulations. AREAS COVERED: This review covers the advantages, disadvantages and applications of TMAs for biomarker research. Research literature and reviews of TMAs and quantitative image analysis methodology have been surveyed for this review (with an AQUA® analysis focus). Applications such as multi-marker diagnostic development and pathway-based biomarker subpopulation analyses are described. EXPERT OPINION: Tissue microarrays are a useful tool for biomarker analyses including prevalence surveys, disease progression assessment and addressing potential prognostic or predictive value. By combining quantitative image analysis with TMAs, analyses will be more objective and reproducible, allowing for more robust IHC-based diagnostic test development. Quantitative multi-biomarker IHC diagnostic tests that can predict drug response will allow for greater success of clinical trials for targeted therapies and provide more personalized clinical decision making.

Personalizing rare disease research: how genomics is revolutionizing the diagnosis and treatment of rare disease.

A decade after the complete sequencing of the human genome, combined with recent advances in throughput and sequencing costs, the genetics of rare diseases has entered a new era. There has now been an explosion in the identification and mapping of rare diseases, with over 10,000 exomes having been sequenced to date. This article surveys the progress and development of technologies to understand rare disease; it provides a historical overview of traditional techniques such as karyotyping and homozygosity mapping, reviews current methods of whole-exome and -genome sequencing, and provides a future perspective on upcoming developments such as targeted drugs and gene therapy. This article will discuss the implications of these methods for rare disease research, along with a discussion of the success stories that provide great hope and optimism for patients and scientists alike.

Quantitative in situ cancer proteomics: molecular pathology comes of age with automated tissue microarray analysis.

Tissue microarrays provide a high-throughput method for assessing a large number of samples by incorporating small cores of tissue into an array that can fit onto one microscope slide. Analyses of tissue microarrays were previously limited by semiquantitative protein expression analysis using brown stain (chromagen-based) methods. These methods are imperfect for protein expression analyses because of a smaller dynamic range and decreased ability for multiplexing many markers, as compared with objective in situ quantitation of protein expression in tumor samples with fluorescence microscopy by a new technology called Automated Quantitative Analysis (AQUA™). By using AQUA analysis, tissue microarrays can serve a unique role as both a discovery tool and as a validation tool for nucleic-acid expression profiling-based target discoveries with results equivalent to enzyme-linked immunosorbent assay quantitation. The identification of novel prognostic markers can identify subsets of patients at high or low risk upon diagnosis, as well as new targets for potential future therapeutic development or metastatic disease treatment decisions. Thus, AQUA provides an unparalleled opportunity to advance personalized medicine through its ability to multiplex, quantitate and localize in situ protein expression.