The Rothberg Institute for Childhood Diseases is a nonprofit organization founded in 2001 for the advancement of research and technology to better understand and find cures for Tuberous Sclerosis Complex (TSC) in children. The Rothberg Institute (TRI) performs research in its Guilford, CT lab along with promoting development through partnerships with organizations like the TSC Alliance, Yale University, Harvard University and The Fox Chase Cancer Center.
Currently, The Rothberg Institute is engaged in a wide genomic study to reveal new genetic markers of Tuberous Sclerosis along with a cellular motility screen aimed at identifying new drugs capable of limiting the spread of TSC tumors. This cell screen involves the investigation of an immortalized cell line from a TSC tumor which was developed at The Rothberg Institute. As a control, TRI derived a rescue cell line by transfecting the tumor cells with the wild-type gene for Tuberous Sclerosis Complex, TSC 1 and TSC 2. Using advanced microscopy software developed at TRI and a luminescent reporter, cells that have been screened against various compounds are analyzed for motility inhibition and cell viability.
TSC cells and the rescue cells are grown in separate 96-well plates till confluence. Each well is then subjected to a light "scrape" producing a thin line in the cells. The wells are then marked for image orientation and high resolution microscope images are taken of each well. After a fresh fluid exchange, the cells are treated with 1ul of either single molecules or organic extracts along with various motility and cell death controls. Over the course of 48 hours, the motility of the cells is recorded through a series of high resolution images. At the end of the growth cycle, a final round of images is taken followed by treatment with a luminescent reporter of cell viability designed to determine if wells with inhibited motility have simply died. Compounds showing inhibition of motility in TSC cells but not rescue cells while maintaining control levels of viability signal are considered exceptional hits