EUPSA - Circular RNA profiles isolated from amniotic fluidscan distinguish between CDH survivors and non-survivers - Richard Wagner

So our last ESA presentation comes from Doctor Richard Wagner from Germany. His presentation is titled Circular RNA Profiles Isolated from Amniotic fluid that can distinguish between CDH survivors and non-survivors. And then we will have Doctor Udo Rohle from, uh, also Germany in Frankfurt, uh, give his take on this manuscript. Thank you very much for the opportunity to present our work today at Stay Current Best of the Best. And I'm gonna talk about how we studied circular RNA profiles in amniotic fluid samples and how they can distinguish between CDH survivors and non-survivors. Prenatal diagnosis and prognostication of CDH cases relies currently only on imaging modalities, but sensitivity for CDH cases lies around 70% and remains imperfect in terms of family counseling and prognostication. Therefore, we try to find and establish biomarkers for early prenatal prediction of outcomes in CDH cases. One type of molecule with high potential for biomarkers that are organ and development specific are circular RNAs. Circular RNAse is an RNA species with a circular shape that is highly stable in biofluids and again has potential as biomarkers in several human disease contexts. As a first step towards biomarker discovery in CDH cases, we profiled circular RNAs in human fetal CDH lungs and compared these profiles to control fetal lungs from patients that didn't have a disease related to the lung. Therefore, we isolated RNA from uh fetal lung tissues that were obtained from pathology, uh pathology specimen, and we profiled circular RNAs with use of a microarray, and then we analyzed our data with bioinformatics. What we found is that circular RNA profiles are significantly altered between CDH and control lungs at two different developmental time points. So, here you can see volcano plots highlighting circular RNAs which are in total 12,000, and each dot represents a circular RNA. So dots that go to the left are down regulated in CDH and dots that go to the right are circular RNAs that are up-regulated, and the ones in red are significantly changed between CDH and control lungs. So we concluded that circular RNAs are different between CDH lungs and control lungs. Next, we applied an algorithm to test whether based on our circular RNA profiles, we could distinguish CDH from control lungs, and the answer is yes, our algorithm could cluster CDH lungs and controls into two separate groups, and that was true for both time points of investigation. As a second step towards establishment of a prenatal biomarker, we now used amniotic fluid samples that were obtained from CDH cases undergoing fetal, so prenatal tracheal occlusion of the fetal trachea. And now we compared circular RNA profiles between patients that later survived CDH and non-survivors. It's important to say that at the time point of amniotic fluid sampling, there was no difference between the two groups. Observed over expected lung to head ratio was similar between survivors and non-survivors. Also, gestational age, liver herniation, or birth weight did not differ between the two groups. However, when we looked at circular RNA profiles isolated from amniotic fluid samples between survivors and non-survivors, we found that circular RNA profiles can distinguish between the two groups, indicating that they have some prognostic value that is superior to the current imaging modalities. Our goal is to validate these biomarkers in amniotic fluids in a larger and independent clinical cohort and define 2 to 3 markers that can predict outcomes in CDH cases, and also we want to test whether this marker can be isolated from maternal blood samples instead of amniotic fluid. This would make an ideal biomarker for prenatal CDH outcome prediction. Thank you very much. I'm very happy to take any questions. Thank you very much to the best of the best team and best greetings from ISA to all of you. And thank you, Richard, for presenting this, uh, very important study. And Todd, I can assure you this is the 3rd, the 3rd game-changing paper which comes from EUSA. Um, Richard, um, you have to expect the question. We're all talking about, uh, success of translational medicine and, um, And we, we, we can actually see, uh, see it on the, on the, on the horizon. So what, what, what would you expect, um, what are your next steps? When will this beautiful marker available? Just taking a bit of blood from the mother and we know there is CDH or no. And if there is CDH, we actually know about the outcome. Can you elaborate on your next steps, please? Yeah. Thank you, uh, Professor Role for that question and thank you for staying current to organizing this beautiful event. Um, of course, the COVID pandemic didn't help us with the study because our next steps involve more patient recruitment, more CDH cases, and ideally different, uh, different severity scenarios, so that we can have increased numbers in each group of our cohorts. And as a first next step, we have to validate what we find in that initial screening experiment. So what we did for now was we screened all the circular RNAs that can be detected in one sample, and now we have to use bioinformatics and additional samples to narrow down the most um significant markers that can distinguish between different outcomes. So really our main goal is, and we just got funded for this by the Canadian Institutes of Health Research in a um multi-center uh fashion study to increase sample size, and then, yeah, statistically narrow down the most promising markers. So that's the next steps. So you are now going into, in, in, in the sort of a clinical trial, is this correct? Yeah, definitely. That's, that's what I would say. We got one minute left. Keep going. Yeah, yeah, OK, um, yeah, I, I thought, I thought that was the the final. I know that's for us internally. That means one minute. OK, um, yeah, so, um, currently we are, uh, working with different centers that have large, uh, CDH. Cohorts, um, to, to really get more cases. And, and if you want to say that this is a clinical trial, I mean, we're not testing any, any medication on these patients, but for sure, we're in big clinical cohorts to, to determine the perfect marker, um, yes. Maybe, maybe a difficult question. How do you deal with uh patients which have another uh malformations, uh, addition to CDH? Yeah, that's a perfect question. So, as for now, we're not powered to stratify these um cases of like syndromic CDH versus non-syndromic and then the different severities, but definitely we would have to look into that quite, quite uh well to see whether we just detect any malformation, but we do believe that with what we've done before, circular RNAs and CDH lungs, that we have a disease-specific marker ultimately. All right, this was, thank you. I'm uh pretty blown away here. Look, I went to Utah, I think a couple of years ago, and had the time of my life. The papers were great. The, the, the parties were even better. Uh, but I will tell you, seeing these three papers reminds me I need to go back. Um, phenomenal that otherwise I would have missed these, never seen these. These are 3 game-changing papers. Uh, so thank you all for presenting these. Um. Uh, Richard, there are a ton of comments. Uh, if you could please go and address some of those, a lot of, a lot of really great discussion points.