An ET-CURE pilot project supporting undergraduate training in cancer research, emerging technology, and health disparities.

The National Cancer Institute's Center to Reduce Cancer Health Disparities has created pilot training opportunities under the "Continuing Umbrella of Research Experiences" program that focus on emerging technologies. In this pilot project, an 18-month cancer biology research internship was reinforced with: instruction in an emerging technology (proteomics), a transition from the undergraduate laboratory to a research setting, education in cancer health disparities, and community outreach activities. A major goal was to provide underrepresented undergraduates with hands-on research experiences that are rarely encountered at the undergraduate level, including mentoring, research presentations, and participation in local and national meetings. These opportunities provided education and career development for the undergraduates, and they have given each student the opportunity to transition from learning to sharing their knowledge and from being mentored to mentoring others. Here, we present the concepts, curriculum, infrastructure, and challenges for this training program along with evaluations by both the students and their mentors.

Comparison of methods for profiling O-glycosylation: Human Proteome Organisation Human Disease Glycomics/Proteome Initiative multi-institutional study of IgA1.

The Human Proteome Organisation Human Disease Glycomics/Proteome Initiative recently coordinated a multi-institutional study that evaluated methodologies that are widely used for defining the N-glycan content in glycoproteins. The study convincingly endorsed mass spectrometry as the technique of choice for glycomic profiling in the discovery phase of diagnostic research. The present study reports the extension of the Human Disease Glycomics/Proteome Initiative's activities to an assessment of the methodologies currently used for O-glycan analysis. Three samples of IgA1 isolated from the serum of patients with multiple myeloma were distributed to 15 laboratories worldwide for O-glycomics analysis. A variety of mass spectrometric and chromatographic procedures representative of current methodologies were used. Similar to the previous N-glycan study, the results convincingly confirmed the pre-eminent performance of MS for O-glycan profiling. Two general strategies were found to give the most reliable data, namely direct MS analysis of mixtures of permethylated reduced glycans in the positive ion mode and analysis of native reduced glycans in the negative ion mode using LC-MS approaches. In addition, mass spectrometric methodologies to analyze O-glycopeptides were also successful.

Overview of the pipeline for structural and functional characterization of macrophage proteins at the university of queensland.

This chapter describes the methodology adopted in a project aimed at structural and functional characterization of proteins that potentially play an important role in mammalian macrophages. The methodology that underpins this project is applicable to both small research groups and larger structural genomics consortia. Gene products with putative roles in macrophage function are identified using gene expression information obtained via DNA microarray technology. Specific targets for structural and functional characterization are then selected based on a set of criteria aimed at maximizing insight into function. The target proteins are cloned using a modification of Gateway cloning technology, expressed with hexa-histidine tags in E. coli, and purified to homogeneity using a combination of affinity and size exclusion chromatography. Purified proteins are finally subjected to crystallization trials and/or NMR-based screening to identify candidates for structure determination. Where crystallography and NMR approaches are unsuccessful, chemical cross-linking is employed to obtain structural information. This resulting structural information is used to guide cell biology experiments to further investigate the cellular and molecular function of the targets in macrophage biology. Jointly, the data sheds light on the molecular and cellular functions of macrophage proteins.

1st NCI annual meeting on Clinical Proteomic Technologies for Cancer.

The National Cancer Institute of the US National Institutes of Health established a Clinical Proteomic Technologies Initiative for Cancer (CPTI) in 2006. The first annual meeting organized by the CPTI program provided up-to-date information on the research activities and achievements at its first anniversary of this program. Presentations were made by leaders from the five centers nationwide of the Clinical Proteomic Technology Assessment for Cancer (CPTAC), and other principal investigators funded by the CPTI.

Genomics and proteomics methodologies for vulnerable populations research.

This chapter describes common genomic and proteomic methods and their application to the study of vulnerable population groups. The International HapMap project is discussed in relation to unique Haplotype single nucleotide polymorphisms (htSNPs) in population groups. In addition, studies, which have used these methods to investigate aging, ethnic, and racial specific conditions, as well as psychiatric diseases, are reviewed. Advantages and limitations of various genomic and proteomic approaches are discussed in relation to population admixture and sample selection.

[Proteomics of breast cancer: From differential to functional analysis]. / Protéomique du cancer du sein: du différentiel au fonctionnel.

From differential analysis to identify biomarkers, to functional analysis for finding new therapeutic targets, proteomics bring new comprehensive information for a better understanding of the molecular basis of oncology and new perspectives for the clinic. However the major limitation of proteomic investigations, more generally of post-genomic approaches, remains the molecular and cellular complexity of the mammary gland that is still a major challenge.

Focusing in on structural genomics: the University of Queensland structural biology pipeline.

The flood of new genomic sequence information together with technological innovations in protein structure determination have led to worldwide structural genomics (SG) initiatives. The goals of SG initiatives are to accelerate the process of protein structure determination, to fill in protein fold space and to provide information about the function of uncharacterized proteins. In the long-term, these outcomes are likely to impact on medical biotechnology and drug discovery, leading to a better understanding of disease as well as the development of new therapeutics. Here we describe the high throughput pipeline established at the University of Queensland in Australia. In this focused pipeline, the targets for structure determination are proteins that are expressed in mouse macrophage cells and that are inferred to have a role in innate immunity. The aim is to characterize the molecular structure and the biochemical and cellular function of these targets by using a parallel processing pipeline. The pipeline is designed to work with tens to hundreds of target gene products and comprises target selection, cloning, expression, purification, crystallization and structure determination. The structures from this pipeline will provide insights into the function of previously uncharacterized macrophage proteins and could lead to the validation of new drug targets for chronic obstructive pulmonary disease and arthritis.

The HUPO Brain Proteome Project--no need to hurry?

The HUPO Brain Proteome Project (HUPO BPP) is dedicated to the analysis of the brain proteome and has initiated two pilot studies in order to elaborate a standardised system for data collection and reprocessing. Samples of mouse brains (different developmental stages) and human brain tissue (biopsy and post-mortem samples) were shipped to different laboratories in Europe, Asia and the US that were invited to identify as many proteins as possible using their own approaches. In addition, a centralised data reprocessing strategy has been elaborated in an iterative way to generate highly reliable lists of identified proteins. This consortium could be a good example for a standardized proteomics workflow.

Clinical infrastructures to support proteomic studies of tissue and fluids in breast cancer.

The Danish Breast Cancer Cooperative Group (DBCG) was established in 1977 with the aim to ensure optimal breast cancer diagnostics and therapeutic modalities on a nationwide basis. DBCG was organized in such a way so it represents a broad interdisciplinary collaboration with established clinical databases and biobanks. This review summarizes the infrastructures, such as those of the DBCG, that are required to facilitate translational research studies aiming at further diagnostic and therapeutic improvements through interactions directed at prevention, early diagnosis, and treatment of primary breast cancer.