 I'm gonna move ahead with the rest of our resident presentations. Next, before I do that, though, there's some thank yous that we need to make. First of all, some of our sponsors, AMO and Allergan, without their help, it'd be difficult to put this on and have nice things to eat and breaks to have and we're grateful that they have agreed to support us. Probably more importantly, Elaine Peterson, along with Geno, Megan and Chandler, although Alan, Barbara's, my name's, are up here on this. Elaine is the one who really does this behind the scenes, makes it happen, and she also couldn't do it without Geno, Megan and Chandler doing this, so we all ought to give them a round of applause. Next up, we have one of our first year residents, Lee Ferguson. Lee's talk, if you're following your program, has changed. It has something to do with Baby Lam's genetics and our OP modeling, and I'll let him explain the finer points of that first. Good afternoon, everyone. So initially, I was going to talk about my previous work with stem cell research and gene therapy, but I've actually was blessed to be able to work or at least had the opportunity to work on two new topics, specifically the topic of Rosa and Baby Lam ROP study. And so the title of my talk is Rosa and the Baby Lam, Tale of Genetic Syndrome and ROP Modeling. So again, there's two talks in one. So the first we'll talk about the Rosa project, and then the second talk will be about the Lam model for ROP. So what is Rosa? So Rosa is actually an acronym, and what it is, it's an inherited syndromic retinal dystrophy, and it includes retinal dystrophy, optic nerve edema, splinomegaly, anhydrosis, and headache, the migraine type. So this was first reported actually with our group here with a case series of a mother and two daughters. It's an autosomal dominant pattern, and so as you can see here at the pedigree, first started in the mom and then the two daughters in the third generation, and then even now there's a fourth generation with the granddaughter, and it was identified as a single gene defect, specifically the alpha kinase gene one defect, or gene at least. And so in terms of clinical phenotype, early on these patients typically have this inflammatory response, actually. They then lead to an optic nerve edema, which then leads to further vision change. Typically within the first decade of life you see the inflammatory changes as well as the optic nerve edema, and then in the early teens, that's when you start to get the vision change. Later on in the second and third decade you start to see this retinal dystrophy and then the systemic disorders. In terms of ethnicity, this has been primarily classified in non-Hispanic Caucasians. So this is a representative patient from that initial study. This would be one of the daughters at age 14, and of course this is an FAA, and so what you're looking at is from left to right in the top two panels and going down is just a time series of the FAA. And so as you can clearly see that there's this leakage to the optic nerve head, but there's no CME nor is there a CNV. And then looking at the multifocal for this same patient, you can see that there's clearly a decline in macular function specifically in the right eye. This is just a fundus photo at a later age. And again, this is the time frame where they start to develop this retinal dystrophy. You can see that not only is there that edema, but you can see that there's this surrounding, at least within the macula, there is macular change signifying retinopathy or dystrophy. And again, the left eye. And so what is alpha kinase gene? It was discovered via next generation sequencing using whole exome as well as genome sequencing. This, the information that was gained was through multi-generational cohort study of families, not only within the nation, but also internationally. So this has definitely been expanded from the earlier studies with the case series. In terms of the tissue expression, it's primarily seeing or three areas that have been found to or this protein has been found in is the eyes, specifically RPE, the optic nerve, as well as the PR inter-segment in the spleen, as well as in the sweat glands, specifically myelopithelial cells. In terms of cellular expression, the protein has been seen in centrosomes, as well as spindle poles and the base of the primary cilia. Now, going into the gene mutation, this has been classified as a missense mutation in this alpha phosphorylurid kinase protein. It's specifically the threonine to methionine mutation at position 237, and it's specifically, it's a highly conserved, the gene itself is a highly conserved gene and it's located on chromosome four. So it's found within multiple vertebrate groups. Now, in terms of the mechanism of disorder, the thought is that it actually affects cell cycle in the sense that it affects the centrosome and it affects ciliary dysfunction. In terms of the kinase ability, that kinase functionality has also been disrupted and so post translational changes have been noted as well. Retinal dystrophy is also the concern here and of course we're talking possibly the centrosomal ciliary function that's at effect here. And then of course the RPE polarity has been dysregulated in relation to this alpha kinase gene mutation. Now, my role in this project is specifically to look at the downstream effects of this kinase mutation. Where is this mutation, where is the gene specifically now acting now that it's been altered in terms of functionality? So the best ways that we want to utilize or at least figure this out is to utilize the kinase assay to look for altered function. Also possibly radio labeled kinase assays to see where this protein is now specifically acting. We know that it acts at centrosomes, it acts at the cilia, but there may be other sites within the cell that have not been characterized. And so it's important to find out where this protein that specifically this altered protein has an effect. So that's the first project. The second project is the LAM model for ROP. And so as we all know, ROP is a leading cause of childhood blindness despite our understanding of treatment and the pathophysiology. And in terms of the recent epidemic of ROP, it's not as a result of what used to be the case of high oxygen tensions, it's more about how these infants are now living longer or at least we're able to keep these premature babies alive sooner or more so than we were able to in the past. And so how do we address figuring out how to deal with ROP? We have animal models. Animal models are important because they mimic the disease process. It's important in understanding the pathophysiology as well as the treatment. And so one of the most noteworthy animal models is the rat model. It's a model of retinal neovascularity. And so it actually modulates the functions of the fluctuations in oxygen in newborn infants. And so it's called the oxygen induced retinopathy model. It again recreates this peripheral retinal avascular zone. And with the avascular zone, this is considered the hypoxic area in the retina. It's similar to what happens in the ROP situation. It then is followed by this vasoproliferative phase where you have the vessels growing at the avascular retinal junction. Now, this mimics again, this fluctuations in oxygen levels as experienced by the infants when they're placed in these incubators. And so with the OIR model, the idea is that you wanna fluctuate between 50% and 10% oxygen. And so it's called a 50 to 10 ratio OIR model. However, this doesn't exactly translate to what's seen at the oxygen level for humans. But it is again, a good model for retinal neovascularization. There have been other models that are noteworthy as well. There's the mouse OIR model as well as the beagle OIR model. And so with the OIR phases in relation to ROP stages, what we're looking at are three specific phases. Phase one, of course, is this vasoproliferative phase. And this is mainly a feature of the mouse OIR model. And then again, this is supposed to be representative of ROP stage one or two. Then you have the vasoproliferative stage. This is more synonymous to what would be seen in stage three of ROP with plus disease. And then you have the phase three, which is the fibro-vascular change as well as retinal detachment. And this would be more of a ROP stage four or five. Now, what are the problems associated with current animal models for ROP? Well, humans don't have the specific phase pattern that the animal models go through. And in terms of reproducibility, it's hard to reproduce this specific avascular retina and vasoproliferative in humans. So that's not specifically translatable to humans. Not all stages of ROP are achieved with the current animal models. Neither the rat nor the mouse are able to get to this phase three where you have more of the, or the phase three where you have stage four and five of ROP. The Beagle model has been able to produce retinal folds, but that's about it. Not the detachment that you would see at stage four or five. And in terms of the current model, even though that it's a great model for retinal neovascularization, it is not a premature model. These animals are newborn animals. And so we need something that actually mimics what happens with premature humans. So that leads to the LAM model, an actual model for ROP, as well as a true model for prematurity. It was first discovered by Dr. Albertine and this model specifically was designed to be studied for bronchopulmonary dysplasia. And so with the aid of Dr. Hartnett, we were able to utilize this because of the prematurity similarities. LAMs expressed as abnormality in IGF-1. IGF-1 is important because it's been known to show, it's been important for human growth or infant growth. It shows that with the absence of it, there may be large peripheral avascular retinas of areas. There's an increase of ROP as associated with the lack of IGF-1. And another reason why the LAM model is specifically useful is that it actually has ventilation scenarios which are similar to what's seen in the NICU setting. And of course, there's this neurocognitive concern that's associated with premature infants. And so with the LAM study, they've been able to show that these premature LAMs do have this CNS-abdominal analysis. So for all these reasons, utilizing LAMs for an ROP model makes sense. And so what is my goal in all of this? What are my aims in terms of this LAM study? Well, my goal is to perform specific OCT analysis and the idea is to monitor for structural changes which are consistent with ROP in these premature LAMs. Also, I want to be able to assess and correlate OCT changes with these CNS findings. Again, these LAMs do present with some CNS disorders and so can we see OCT findings which will correlate with that is a question to be asked. So we would specifically look for biomarkers for these neurological changes. That's pretty much it for my two topics. I want to thank the people who were involved with the Rosa Project, Dr. Williams, Dr. DeGree, as well as the individuals involved with the LAM project, Dr. Hartnett, Dr. Albertine and the rest of the Hartnett lab. Thank you. That was a very nice topic. I'd like to ask what changes you would expect to see on the OCT in the LAM model, our case, most things like that affects the periphery. Would you be in any periphery with OCT? So you can image the periphery, although it may be a little bit harder with these animals since they don't typically stop to maintain a good fixation. But the idea is to look for intral retinal changes. You can see intral retinal layers changes. You can also see cystic possibly changes in the retina. There are areas of the receptors that might start to change as a result of the OIR application. Thank you very much. Thank you. Thank you.