MD TP25: Generation of phage libraries from patients with systemic scleroderma to select scFv antibodies against angiotensin 1 receptor and endothelin A receptor

Systemic scleroderma is a disease characterized by extensive fibrosis of the skin and internal organs, including kidney and lung. Recent evidence suggests that functionally active autoantibodies against angiotensin 1 receptor (AT1R) or endothelin A receptor (ETAR) play a key role in the pathogenesis of this disease. This hypothesis is supported by the observation that drugs inhibiting both receptors, angiotensin-converting enzyme inhibitors or endothelin antagonists, have a positive effect on disease progression. The aim of the proposed project is to generate antibody libraries from patient materials to select and reconstruct functionally active autoantibodies. Ultimately, this will aid to investigate the pathogenic role of these autoantibodies. For this project, a set of patient samples has already been collected for preparation of RNA, which will be used to amplify antibody sequences and construct large libraries of single chain variable fragments (scFv). From these libraries, scFvs specific for AT1R, ETAR or both receptors will be selected by panning on CHO cells transfected with respective receptors. Based upon scFv sequences, full-length antibodies can be reconstructed recombinantly to evaluate their functional properties in in vitro and in vivo models. This project will be the basis for a large number of other projects that will help to improve our understanding of systemic scleroderma pathogenesis, and that will lead to novel diagnostic and therapeutic options for patient care.