Fluorescence assay for characterization of regulators of the α2β1-integrin receptor.
Integrins are attractive markers and targets for various diseases. The fluorescence assay of the invention can be used to identify regulators of integrins and determine their influence on integrin expression and localization.
Integrins are important adhesion receptors on the surface of cells, where they interact with a variety of extracellular molecules. Integrins are therefore involved in important cellular processes such as signal transduction, cell migration or differentiation. The availability and functionality of integrins at the cell surface depends not only on their expression level, but also on their transport from the cell interior to the surface, the residence time at the surface as well as the internalization and ultimate degradation of the receptor.
The human integrin protein α2β1, in which integrins α2 and β1 assemble into a heterodimer, is a receptor for laminin, collagen and fibronectin. α2β1 is involved in numerous processes, including hemostasis, organ morphogenesis, angiogenesis or platelet aggregation. In addition to their diverse biological functions, integrins are also associated with various diseases and are therefore attractive therapeutic targets. In this context, altered cell adhesion and cell migration are hallmarks of abnormal cellular processes, especially in the development and metastasis of cancer cells. Accordingly, integrins, especially aberrant expression patterns and behaviors, can be used in diagnostics as indicators for the occurrence of various diseases. In addition, α2β1 receptor is also known to act as a potent tumor suppressor; accordingly, the receptor is of interest in the prevention and treatment of cancer. Despite these various aspects, quite little is known about different biological aspects of the molecule, especially about the processes between production of the protein inside the cell, its presentation on the cell surface and what happens after internalization of the receptor.
Scientists at Heidelberg University have developed a fluorescence microscope-based assay for the systematic identification and investigation of potential regulators of integrins, in particular of the α2β1 receptor. With the method, it is possible to determine and investigate genes, gene products and other modulators of any kind, i.e. of proteins, nucleic acids and/or chemical agents. This is achieved by fluorescence spectroscopic comparison of the amount of detectable integrins in the normal state as well as after exposure to a modulator that affects the expression, localization and/or trafficking of the integrin. The method according to the invention is fast, easy to automate, has a high throughput and is characterized by reliable and meaningful results.
- Integrins as suitable markers (for e.g. cancer) and attractive therapeutic tragets
- Versatile application in all diseases related to integrin-mediated cell adhesion, such as cancer or thrombosis
- Method can be performed quickly, easily and reliably
- Can be automated for high-throughput applications
Fields of application
The method is therefore suitable for the diagnosis and therapy of a wide range of diseases related to integrin-mediated cell adhesion. Possible fields of application include cancer, thrombosis, arteriosclerosis, strokes, disorders of wound healing or arthritis.