Individual genetic findings associated with complex diseases are unlikely to fully explain their substantial impact or provide new comprehensive insights into disease pathogenesis. These also lack ...the comprehensive data much needed for development of new effective drugs in majority of the disease cases in a population. In fact multilevel etiologic factors underlie almost all human diseases, including: environmental causes, epigenetic factors, DNA mutations, amplifications, and deletions, RNA expression levels, protein (translation, post translation modification, localization) and combinations thereof. Each individual might consist of different combinations of these multiple etiologic factors. Integrative evaluation of all these modifications will shed light on the whole identity of the disease and the underlying molecular mechanisms. Until now it was inconceivable to have a full grasp of such a complex etiology. Microarrays enable us to interrogate the individualized various factors (DNA, RNA and protein content) involved in disease state on genome-wide scale simultaneously and expeditiously in single cell or the tissue of interest (Figure 1). The new disciplines of microarray studies in combination hold the promise of effective, current, and comprehensive understanding of complex diseases and may be a good approach for reducing the costs and time lines associated with discovery and efficacy improvement of therapeutic drugs. In the future, through utilizing the colossal amount of microarray data findings, defining the structure, function, and dynamics of entire biological pathways and cellular networks under various physiological states, and the development of robust and efficient methods for analyzing and interpreting high dimensional data, it will be possible to connect combination of experimental results with individualized disease state. This will facilitate precise diagnosis prognosis and therapy.
Microarray technology has its roots in high-throughput parallel synthesis of biomacromolecules, combined with combinatorial science. In principle, the preparation of arrays can be performed either by ...in situ synthesis of biomacromolecules on solid substrates or by spotting of ex situ synthesized biomacromolecules onto the substrate surface. The application of microarrays includes spatial addressing with target (macro) molecules and screening for interactions between immobilized probe and target. The screening is simplified by the microarray format, which features a known structure of every immobilized library element. The area of nucleic acid arrays is best developed, because such arrays are allowed to follow the biosynthetic pathway from genes to proteins, and because nucleic acid hybridization is a most straightforward screening tool. Applications to genomics, transcriptomics, proteomics, and glycomics are currently in the foreground of interest; in this postgenomic phase they are allowed to gain new insights into the molecular basis of cellular processes and the development of disease.
Background: Matrix-metalloproteinases (MMP) are involved in a broad spectrum of physiological processes. Moreover, they also
play a key role in tumour invasion and metastatic spread. The induction of ...MMPs is mediated via the extracellular matrix-metalloproteinase
inducer (EMMPRIN). EMMPRIN is expressed in a variety of epithelial tumours, but expression in non-Hodgkin lymphomas has not
been studied yet. Materials and Methods: Therefore we studied 201 non-Hodgkin lymphomas (NHL) for EMMPRIN expression by immunohistochemistry
using a newly developed tissue microarray (TMA). This new approach to TMA-technology entails the great advantage that areas
of interest (for instance with high tumour cell content) are selected by means of serial sections, thus maintaining appropriate
samples of each case on the array. The samples were evaluated with regard to the number of positive tumour cells and staining
intensity. Results: All specimens on the arrays contained a sufficient amount of tumour cells. Immunohistochemistry yielded
satisfactory results in 196 out of 201 cases. EMMPRIN was expressed in a significantly higher number of tumour cells in high-grade
NHL compared with low-grade NHL. Furthermore, the staining intensity was significantly stronger in high-grade NHL. Conclusion:
We report on a new type of TMA that allows effective parallel analysis of a large number of tissue samples. Our data indicate
that the expression of EMMPRIN is strongly associated with a more aggressive lymphoma type. However, additional studies are
required to elucidate the role of EMMPRIN in the tumour biology of NHL. Possibly, EMMPRIN could be a new target in the therapy
of NHL.
Morphological sciences have recently experienced a significant technological breakthrough that offers new opportunities for cell and tissue imaging in situ but also raises new challenges to ...pathologists, who must adapt to a rapidly evolving environment. New partners, such as cell and molecular biologists, have provided pathologists with highly powerful tools for cell and subcellular imaging. They include: (a) the adaptation of techniques derived from molecular biology and cytogenetics, (b) the development of new microscopic tools, such as confocal microscopy, and (c) the emergence of new preparative techniques, such as microdissection or tissue arrays. However, recent technological progresses in various fields, from endoscopy to genomics, also raise new challenges to pathologists. Pathologists must therefore be prepared to redefine their area of expertise: this will be achieved through a continuous collaboration with all the partners involved in cell and tissue imaging and analysis but also by emphasising the importance of the informations provided by cell and tissue imaging in situ.
There has already been a ‘molecular’ revolution in pathology. Demonstrating transcription of specific single genes or small gene sets and their protein products by
in situ hybridisation and ...immunocytochemistry is routine in diagnostic and experimental pathology. A perhaps-greater revolution is imminent with the application of more recently established and emergent technologies in pathology. These include new approaches to polymerase chain reaction (PCR); simultaneous studies of multiple genes and their expression using oligonucleotide and cDNA arrays; serial analysis of gene expression (SAGE); expressed sequence tag (EST) sequencing, subtractive cloning and differential display; high-throughput sequencing; comparative genomic hybridisation, multiplex fluorescence
in situ hybridisation (FISH) (spectral karyotyping); reverse chromosome painting; knockout and transgenic organisms; laser microdissection and micromachining; and new methods in bioinformatics, ‘data mining’ and data visualisation. Molecular methods will profoundly change diagnosis, prognosis and treatment targeting in oncology and elucidate fundamental mechanisms of neoplastic transformation. Individual susceptibility to specific diseases will become assessable and screening will be refined. The new molecular biology will be most fruitful in partnership with classical approaches to pathology: the expectation that molecular methods alone will answer all pathological questions is unrealistic. A further challenge for the biomedical community in the ‘genome era’ will be to ensure that the benefits of these sophisticated technologies are enjoyed globally.
Die Arbeitsgruppen Chipdiagnostik und Bioinformatik der DGKL haben sich bei ihrer Jahrestagung 2008 mit Anwendungen der arraybasierten Technologie in der Pathologie beschäftigt. Hauptthemen waren ...neue Entwicklungen auf dem Feld der Gewebematrices oder Multigewebeblöcken (engl. tissue arrays), Anwendungen der multiparametrischen Analytik in der epigenetischen Regulation der Genaktivität, sowie die arraybasierte vergleichende genomische Hybridisierungstechnologie (engl. array comparative genomic hybridization oder Array-CGH). Diese Methoden finden bereits heute besonders in der Charakterisierung von Tumorzellen und -gewebe einen breiten Einsatz und haben die Krebsdiagnostik erheblich vorangebracht. Eine wichtiges Ergebnis der Tagung war die Erkenntnis, dass bei der hochparallelen Analytik die technischen Möglichkeiten der klinischen Anwendung voraus sind. Deshalb besteht ein erheblicher Bedarf vor allem an begleitender klinischer Forschung, um die Anwendbarkeit und die Bedeutung dieser Diagnostik zu präzisieren.
In their annual conference in 2008, the Chip Diagnostics and Bioinformatics Working Groups of the Joint German Society of Clinical Chemistry and Laboratory Medicine dealt with applications of array-based technology in the field of pathology. The major themes of the conference were new developments in area of tissue arrays, multiparametric analysis in epigenetic gene regulation, and array comparative genomic hybridization. These methods are already being widely used in the characterization of tumour cells and tissue and represent a significant advance in cancer diagnostics. One important result of the meeting was the recognition that in the field of highly parallel analysis, the technical possibilities currently exceed our ability to put them to clinical use. For this reason, there is a particularly pressing need for accompanying clinical research in order to better define the uses and importance of these diagnostic tools.
Die Arbeitsgruppen Chipdiagnostik und Bioinformatik der DGKL haben sich bei ihrer Jahrestagung 2008 mit Anwendungen der arraybasierten Technologie in der Pathologie beschäftigt. Hauptthemen waren ...neue Entwicklungen auf dem Feld der Gewebematrices oder Multigewebeblöcken (engl. tissue arrays), Anwendungen der multiparametrischen Analytik in der epigenetischen Regulation der Genaktivität, sowie die arraybasierte vergleichende genomische Hybridisierungstechnologie (engl. array comparative genomic hybridization oder Array-CGH). Diese Methoden finden bereits heute besonders in der Charakterisierung von Tumorzellen und -gewebe einen breiten Einsatz und haben die Krebsdiagnostik erheblich vorangebracht. Eine wichtiges Ergebnis der Tagung war die Erkenntnis, dass bei der hochparallelen Analytik die technischen Möglichkeiten der klinischen Anwendung voraus sind. Deshalb besteht ein erheblicher Bedarf vor allem an begleitender klinischer Forschung, um die Anwendbarkeit und die Bedeutung dieser Diagnostik zu präzisieren.
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