Project 1
David Seldin, MD, PhD |
CLONING AND EXPRESSION IN VITRO AND IN VIVO OF AMYLOIDOGENIC LIGHT CHAINS
Project Leader: David Seldin, MD, PhD Professor of Medicine and Microbiology
The underlying hypothesis for this project is that the immunoglobulin light chain (LC) gene sequence present in the clonal plasma cells in patients with AL amyloidosis is ultimately responsible for the manifestations of disease. The gene encodes a unique protein that is folded, undergoes post-translational modification, associates with accessory proteins, and interacts with target cells and organs. All these properties contribute to amyloid disease, but all are dictated by the underlying primary amino acid sequence encoded by somatic rearrangement of immunoglobulin genes in B cells, and preserved in their plasma cell progeny. To test this hypothesis, we established “ALbase”, a database for amyloidogenic LC sequences from patients, populated it with more than 200 new sequences, and analyzed their predicted amino acid sequence and their structural properties. This database is still under development, and will go "live" in 2007. We cloned LCs into vectors suitable for in vitro and in vivo expression, and developed a transplantation animal model and a successful transgenic model in which AL amyloid is formed spontaneously in vivo. Studies of these animal models are ongoing. We are using them to investigate immunotherapy and gene therapy approaches to treatment of AL amyloidosis.
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