Pilot Project 6

Novel Affinity Reagents for Analysis of Tumor Microvesicles and Their Metabolically Regulated Glycome (competitive renewal of Year 1 Pilot Project)

Project Investigators: Matt DeLisa, Matthew Paszek

Additional Collaborators: Sachdev Sidhu, Wesley Legant

Metabolic reprogramming is a hallmark of cell fate transitions. Cancer cells often acquire a “Warburg-type” metabolic phenotype that is characterized by unusually high rates of glycolysis and lactate production. Frequently occurring oncogenes, including KRas, activated PI3K, ErbB2, and c-Myc, are now recognized to drive dramatic changes in glucose metabolism. Such changes in metabolism alter the flux of metabolites through anabolic pathways, including those that generate building blocks for the sugar-rich outer structure of cells called the glycocalyx. Moreover, metabolic reprogramming also results in the biogenesis and shedding of microvesicles (MVs), which play critical roles in tumorigenesis. The regulation of MV biogenesis and function, its relationship to metabolism and the glycocalyx, and the role of the physical microenvironment in these processes, however, remain obscure. This pilot project renewal seeks to address these gaps in our understanding through the development of novel glycan-binding affinity reagents for fluorescently labeling the tumor MV glycome and specific “glyco-domains” on the plasma membrane from which MVs originate. The proposal builds off of a highly successful Year 1 Pilot Project involving four investigators with expertise in areas that are relevant to the Cornell Center on the Physics of Cancer Metabolism, including glycoengineering and glycan-directed antibody (Ab) development (DeLisa), glycocalyx imaging and functional analysis (Paszek), super-resolution imaging (Legant), and high-throughput selection of synthetic Abs (Sidhu). By combining approaches from glycoscience, Ab engineering, and nanoscale molecular imaging, our project will advance the state-of-the-art in tumor MV research through creation of new technologies and protocols. With these technologies, we will test how metabolic reprogramming in cancer alters the nanoscale structure of the glycocalyx and how these “spatial mutations” change the rates of tumor MV production.