Generation of in Vitro Cellular Models of Lymphangioleiomyomatosis for the Development of Tuberous Sclerosis Therapeutics

摘要

The purposes of this work are three related objectives: to generate human TSC2-/- LAM cell lines; to generate matching inducible isogenic TSC2 knock-in cell lines; and to confirm the absence and rescue of TSC signaling in these LAM lines. We generated 400 immortalized LAM cell lines from two different LAM tissue sources and used immunoblot analysis to determine the loss of the TSC2 protein itself, or function as indicate by phosphorylation of the ribosomal protein S6 in the absence of serum. Despite the fact that the original LAM tissues suggested loss of TSC2 by FISH analysis in a subset of the tissue, none of the newly developed lines appear to be negative for TSC2. We feel, however, that these lines still be valuable to the TSC and LAM communities and we will therefore, disseminate the generated cell lines upon request.