Team:Stony Brook/Description
Background
Cancer is a widespread problem affects many people from around the world. Pancreatic cancer in particular tends to be detected very late and doesn't offer patients much time after discovery. The circulating exosomes, or extracellular vesicles, secreted by cells in the human serum express many surface proteins, one of which is called Glypican-1 (GPC-1). This protein is expressed in under 10% of exosomes in healthy people, but jumps to 50% in people prone to pancreatic cancer, and that number only rises with increasing susceptibility to tumor formation. Thankfully, the expression GPC-1 increases months before the tumor forms. We hope to target this protein and use it for our project.
Motivations for our Project
Since pancreatic cancer is a problem due to its elusiveness in diagnoses, many people find out they have pancreatic cancer too late and prognoses tend to be very negative. Our team is hoping to change that by offering an alternative method for early detection. Current methods involving the biomarker CA19-9 take a few days and only have 71% sensitivity, so we hoped to find a new biomarker that predicted susceptibility to tumor formation. The biomarker we found was Glypican-1. We wanted to a design a method that would allow us to immediately tell if high amounts of GPC-1 were present and could be administered cheaply, but maintain accuracy, so we decided to use yeast as our chassis, as it could easily be replicated if we designed a system that would be able to detect Glypican-1.
Early Detection of Pancreatic Cancer
Our project directly aims to provide a method of detecting pancreatic cancer in humans. We are also using yeast in this project as a chassis for expressing Cripto-1, a membrane protein which binds to Glypican-1 (GPC-1). We are designing a construct including Cripto-1 and a secretion tag to ensure that our protein is secreted to the membrane. We plan to use Cripto-1 antibodies on frozen cells and depending on where the antibodies aggregate, we can determine if Cripto-1 was indeed secreted to the membrane. C-SRC is a tyrosine kinase receptor that is phosphorylated by Cripto-1 in humans. We plan to test yeast cells both with and without C-SRC in the presence of GPC-1 to test if yeast C-SRC actually is phosphorylated by Cripto-1. C-SRC functions through the MAP Kinase pathway. We hope to be able to couple Cripto-1 binding to a reporter, GFP, inside yeast for indication of the presence of the GPC-1 in human blood serum. GFP would be expressed at a basal rate without the binding of GPC-1 to Cripto-1, but if binding occurs, GFP's expression would be augmented and could be measured with a spectrophotometer to determine susceptibility to tumors. This could potentially offer insight into the potential for synthetic biology as an important medical tool in the future.
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