 In our lab we study coronary artery disease or CAD, which is the single largest cause of death worldwide. Many of us don't know that CAD begins in our late teens, early 20s. As a 35 year old former smoker, I'm actually a pretty good candidate myself. But advanced CAD mainly manifests in people 55 years of age and older, and it takes two very distinct forms. In panel A what you see is what we call stable plaques and these are plaques where although the artery has become narrower, the plaque itself is at lesser risk for rupturing and reducing its contents into the lumen. Contrasting with this is what we see in panel B, which are much more dangerous vulnerable plaques and what distinguishes them from stable plaques is the thin fibrous cap which you'll see right above the black arrow. And so in a vulnerable cap when that thin fibrous cap ruptures what happens is what we see in panel C where the contents of the necrotic core spill out into the lumen and these usually form a thrombus which occludes blood flow to the heart muscle leading to a myocardial infraction and unfortunately most often a fatal myocardial infraction. So one of the big big unknowns of cardiology is what exactly causes a vulnerable plaque to go into that ruptured state. If we understand this, this helps us immensely in managing advanced cardiovascular disease. So my specialty is in applying RNA sequencing to the study of cardiovascular phenotypes such as plaque rupture which is what we see here. And one of the things I'm doing to understand this question is I'm sequencing RNA from subjects who died from sudden cardiac death and what sudden cardiac death is a person develops symptoms usually chest pain and then within the next hour they're dead and during autopsy they're found to have a ruptured plaque like the ones we see here which caused the myocardial infraction that led to their dying. So we want to use RNA sequencing as a tool for studying the changes leading up to this event. The experimental design we use for this project is about half pathology and half sequencing. First we collect hearts from the autopsy procedure and then we put these hearts through a pretty detailed protocol to isolate the lesions, characterize them, understand what caused the death and by taking the data from that procedure we have formed a matched set of 10 hearts which had both vulnerable and ruptured plaques and then from 10 other hearts we have 10 stable plaques like the one in panel A which we use as controls. So our next challenge is to sequence the RNA from these plaques and that is unfortunately a little harder than it sounds because the hearts were originally collected not for sequencing but for pathology procedures and the chemicals used for the histology staining usually degrade the nucleic acids pretty badly. So I'm however quite happy because we've been able to recover substantial amounts of RNA in a size range that although it's not perfect is still usable for sequencing. So in panel D what you'll see is relative to the high quality near perfect RNA on the extreme right the other eight samples which are in the two other boxes to the left are the samples we get from these autopsy hearts and as you can see they're pretty fragmented compared to the control sample but the good news here is that if you look at the box in the middle which is RNA that's 200 base pairs or greater in size we still have a substantial amount of RNA in that size range and that's actually what we need to use sequencing on these samples. So I'm really very optimistic that once we have the results from sequencing these we'll be in a pretty good position to understand from the transcriptome point of view what went on that led a stable plaque to go in the direction of becoming vulnerable and even more importantly what happened between the vulnerable plaque and the rupture event that caused sudden cardiac death. If we understand that that gives us a good starting point to be able to manage this disease hopefully lower rates of cardiovascular mortality which as I mentioned are the highest in the world.