Forschungszentrum Borstel
Forschungszentrum Borstel

Medicine

Pathology of the University Hospital Schleswig-Holstein, Campus Lübeck and the Leibniz Research Center Borstel
location Borstel

Clinical and Experimental Pathology

Projects

Pulmonary disorders represent a major medical, biological and economical problem which we are engaged to face as the center of our scientific work and interest.
The Clinical and Experimental Pathology therefore serves as a fully equipped diagnostic facility for different hospitals and practitioners as well covering all aspects of pathology with focus on pulmonary pathology. Excellent diagnostics are one prerequisite for both efficient therapy and outstanding research.
To accomplish this, a widespread variety of histo- and cytopathologic techniques, immunohistochemistry, electron microscopy, telepathology, digital imaging and molecular pathology as well as autopsies are applied. Together with excellently trained personal and experienced pathologists the Clinical and Experimental Pathology serves as a linker between clinical and scientific purposes either in the field of medicine or natural sciences. The expertise of the Borstel Pathology according to occupational pulmonary diseases has been increasingly requested by different professional organizations as well as by other pathologists. From the widespread material, especially from general oncopathology, the diagnostics of pulmonary diseases are profiting. In this context, together with the clinicians, numerous case reports were published. As a center for molecular-pathology in the northern part of Germany we regularly train guest pathologists over longer time spans. Molecularbiological detection of mycobacterial DNA-sequences, in close cooperation with the National Reference Center for Mycobacteria (FZB), as well as the detection of other germs (like EBV, HPV etc.) by the use of archive-material are performed as a routine. Clinical-pathological conferences are regularly carried out.
Numerous interdisciplinary projects therefore could be successfully carried out in the past, presence and future.
Research on pulmonary disorders to date is carried out mostly by the use of model-systems such as cell culture and animal models, which enable functional studies and can be used for the development of novel treatment strategies. Due to the large number of possible pathogenic processes and a variety of different cell types involved a complete simulation of pulmonary diseases in these model-systems can hardly be accomplished.
For these reasons the use of functional model-systems based on human tissues bear great advantages. Histological sections display a kind of snapshot of the complex mix of events taking place in the course of disease.
As a link between cell-culture and animal models the human pulmonary alveolus represents a structural and functional unit during the course of pulmonary diseases. Methods for manipulation and stimulation of tissues in culture - which means functional studies in tissues - are long time known, yet the crucial tools were widely restricted to analyze the effects within the tissues in situ on the molecular level.

To overcome with this, we have developed molecularbiologic applications for the use with the HOPE- (Hepes - glutamic - acid - buffer - mediated - Organic - solvent - Protection - Effect) technique during the last years. This is an alternative fixation leading to paraffin embedded tissues with well preserved morphology together with well preserved and accessible nucleic acids and proteins that allows complete pathological analysis of all types of human soft tissues. Furthermore, it maintains additional positive features relevant to immunohistochemistry and molecular pathology. The so-called HOPE-technique comprises a protection-solution with an organic buffer, acetone as the only dehydrating agent and pure paraffin of 52-54°C melting temperature. Although the exact mechanism of protection has still to be elucidated, it seems rather unlikely that chemical bindings occur during the whole process of fixation which is described and compared with the standard formalin-paraffin technique. Essentially, HOPE-fixed sections show formalin-like morphology.
We have developed protocols to be used with the HOPE-technique in an unambiguous fashion which allow successful DNA and RNA in situ hybridization, PCR and RT-PCR analysis, Western-blot, Northern-blot, microarrays as well as immunohistochemistry with tissues and single cells. The results of these clearly outperform those obtained with conventional fixatives and which yet all have been widely restricted if using archived paraffin materials.
As a result of this, we are now capable to perform proper read out of molecular events taking place within human and other tissues, which allows for functional studies in short term tissue cultures as a model we are currently working on (STST: short term stimulation of tissues). This model is used for our (in situ) studies of non small cell lung cancer specimens as well as in an ex vivo model for infectious diseases.

Application of the HOPE-technique moreover allows for diagnostic purposes as we have shown for the enhanced detection of biomarkers by immunohistochemistry important for treatment and prognosis in cancer patients.

Molecularpathologic subjects like the detection of mycobacterial DNA in paraffin-embedded tissues also substantially profit from the HOPE-technique, which is why we conclude the HOPE-technique to be of great value within the growing field of molecular diagnostics.

Taken together, we follow a straightforward tissue-based attempt to explore complex molecular events taking place in the course of pulmonary diseases aimed at improving diagnostics and developing novel treatment regimens in the future.