Reseach fellows supported by the Boston Biomedical Research Institute Senator Paul D. Wellstone Muscular Dystrophy Cooperative Research Center for FSHD:
Emerson Laboratory
Min Du, Ph.D.
Boston Biomedical Research Institute
64 Grove Street
Watertown, MA 02472 USA
du@bbri.org
Proteomics and molecular biology
MS studies of proteome of FSHD myoblast lines
Dr. Min Du’s research focuses on searching for the FSHD protein biomarkers to achieve a better understanding of FSHD development and progression, and identification of potential pharmacological targets. In complement to the informative microarray analysis of differential mRNA expression, this study will reveal the differential protein expression with information of its posttranslational modifications. The current goal is to develop a comparative proteomics workflow using mass spectrometry (M-S) based approaches for high throughput analysis to discover the FSHD protein biomarkers in a fast and reliable manner. Dr. Du is developing a proteolytic 18O labeling using PALeO (protease activity labeling employed 18O-enriched water) strategy, which can be applied to clinical samples. PALeO is an in-house information system, which is used to characterize proteolytic activities in complex biological samples. Dr. Du is extending this method in 18O-labeling based relative quantitative proteomics. In preliminary studies, Dr. Du used BSA (Bovine Serum Albumin) as the standard protein to test PALeO strategy in M-S based protein quantitation analysis. Compared to literature reports, her preliminary data based on BSA showed highly reproducibility with tighter and more accurate calculated ratios. Dr. Du is applying this workflow to normal and FSHD patient myogenic cells, and then to clinical patient samples. Alternatively, Dr. Du is also developing a SILAC (Stable Isotopic Labeling using Amino Acids in Cell Culture) based strategy for cultured normal and FSHD patient myogenic cells.
Miller Laboratory
Vivek K. Vishnudas, Ph.D.
Wellstone Center Postdoctoral Fellow
Boston Biomedical Research Institute
64 Grove Street
Watertown, MA 02472 USA
vishnudas@bbri.org
Muscle physiology
Role of SIRT1 in differentiation, apoptosis and myogenesis in FSHD
The goals of Dr. Vivek K. Vishnudas’ project are to increase our understanding of facioscapulohumeral muscular dystrophy (FSHD) pathogenesis and develop platforms for drug screening through identification of critical differences between FSHD and normal myogenic cells. The work examines proliferation, differentiation, muscle-specific gene expression, and apoptosis/necrosis and has a particular focus on how oxidative stress can alter these phenotypes. Abnormal susceptibility to oxidative stress is considered to be a property of FSHD myogenic cells. Dr. Vishnudas is devising simple in vitro assays to identify differences between FSHD and normal myogenic cells and also determining which signaling pathways might regulate the aberrant responses of FSHD cells. As an example, his preliminary work suggests that FSHD myoblasts are significantly more susceptible than normal myoblasts to inhibition of differentiation by a high concentration of pyruvate. As he identifies significant differences between FSHD and normal myogenic cells, these differences will be used as assays to identify pharmacological agents that might reverse the abnormal properties of FSHD muscle cells
