 So, Jyoti Dayal is a program director in the Division of Genomic Medicine and she's going to present the concept title, Developmental Genotype Tissue Expression, or DGTEX, is probably what you're going to hear for the rest of the afternoon. Jyoti. Okay. Okay. All right. Well, thank you. Good afternoon, everyone. Thank you, Council and staff, for this opportunity to present to you the Developmental Genotype Tissue Expression Project. Before beginning, I'd like to thank members of the DGTEX working group, staff from both NACRI and NICHD, the National Institute of Child Health and Human Development, both for their support and guidance. We've been working with NICHD on several initiatives and would like to continue our collaborations with them. For today's presentation, I would like to start off by highlighting the accomplishments of the Adult Genotype Tissue Expression Project, lay the groundwork for DGTEX, the preparatory work that's been done thus far, and present the consortium model and budget we would like to propose. We're building off the accomplishments of GTEX as it established a rapid autopsy program recruiting 965 donors from 2010 to 2016. Every gene expression in 53 tissues, including 11 subregions of the brain, provided a new approach to map gene expression, and decoded regulatory regions of the genome. And as a result, published over 2,000 consortium and non-consortium papers, and second only to the cancer genome atlas and data access requests. NHGRI and NICHD co-hosted the Genomic Medicine for Reproductive, Pre-Natal, and Neunatal Health Workshop. One of the recommendations was the creation of a genotype tissue expression project for neonates and children. The workshop summary can be found on genome.gov. So we're hearing a similar theme and support for this idea at other scientific meetings, several strategic planning meetings, and even the World Congress psychiatric genetics meeting. And the question everyone is asking is what about expression in the developing human? The life stages of human development are very different. A child is not a small adult. There are differences that occur during development such as metabolism, regulation of hormones, immunity, drug response, and exposure to environmental toxins to name a few. And so DGTEX would provide that critical information by establishing a resource database to study gene expression patterns across developmental stages. This would help fill key gaps in understanding gene expression, provide insight on functional networks and pathways, and understand how gene expression affects clinical factors such as drug response in early and later childhood. The proposal I'd like to present, similar to GTEX, which cataloged and analyzed transcriptional profiles across multiple tissues in adults. DGTEX would also analyze transcriptional profiles in multiple tissues, but across developmental stages. And this would provide a comprehensive data set from neonatal through adult life stages. The objectives for DGTEX is first to create an atlas of tissue gene expression on both tissues and single cell populations, and would like to emphasize that DGTEX would benefit from the rise of single cell technology to understand how distinct cell types influence each other during the development of organs and tissues. Then to analyze regular square variation of genomic sequencing and its effect on gene expression. And then finally to create and make available a bivank of tissues and data for further characterization. This last point is particularly important to prep samples in such a way to do additional analyses. This is the current study design. We will be recruiting 40 donors for each of these age groups, early postnatal, pre-pubertyl, for a total of 120 donors. We selected these age groups as it represents significant changes in development. Blood samples we've collected for whole genome sequencing on all donors and RNA sequencing will be done on a subset of tissues and single cells. The selection of tissues for RNA sequencing will be defined by what is accessible to collect and the quality of the tissue. While collecting a suite of tissues, this will allow us the ability to do additional analyses such as chromatin accessibility, histone modification, and DNA methylation on those bulk and single cells. We realized that special considerations need to be made, especially for children, one of them being the ethical issues of posthumistinomic research in neonatal and pediatric settings. We requested a biothics consult service from the NIH clinical center to help anticipate and analyze ethical issues that might arise in the prospective research activity. Those recommendations included engaging families and health care staff on how to approach greeting parents about tissue donation and also being mindful of cultural and religious differences in diverse communities. To consider eligibility criteria for participation, to ask questions that are absolutely necessary during consent, and to follow up with the family members maintaining communication and providing general research study results. The other consideration is tissue procurement feasibility in pediatric populations. With the help of our NICHD colleagues, we attended two NICHD funded steering committee meetings and asked experts how feasible it is to obtain multiple post-mortem tissues for research. The two take-home messages are that families are amenable towards organ donation research and that we should be working with existing infrastructures, organ procurement organizations and medical examiners. This is the basic consortium models, and I just want to remind you that the goal of DGTEX is to create a resource database to study gene expression across developmental stages. NICHD will be managing the tissue procurement centers, which will be responsible for recruitment of donors, and we would be building off the inclusion and exclusion criteria that DGTEX developed, pathology of the tissue samples that would be acceptable for collection, and again we would leverage the tissue procurement protocols developed by adult DTEX, collecting clinical data and LC research to assess the attitudes and perspectives of tissue donation from various stakeholders. Tissue alquots will then be sent to the LDAC, the laboratory data analysis and morning center, and NICHD or I will be managing the LDAC and the statistical analysis and data integration, and for today I'll be focusing on the LDAC. Here's a sample of the major activities for each of the LDAC components. The laboratory will purify nucleic acids, perform whole genome and RNA sequencing, and maintain the biobanking facility. The data analysis group will perform basic genotyping and expression analyses, single cell analyses, and prepare the deposition of data sets and repositories, such as the ANFO, ensuring that the data is hosted on the DGTEX portal and integrate data into the UCSC genome browser. The Corning Center will work with the tissue procurement centers and monitor study progress and laboratory performance and prepare statistical and other reports. This is a distribution of funds for each of the solicitations. We're asking council to vote on the DGTEX concept and specifically the LDAC. NICHD council approved the Tissue Procurement Center's solicitation, and the data analysis and integration of DGTEX data have not been developed yet, but will be brought to you at a later council. The total budget will be split between NHGRI and NICHD. Other ICs are being approached for additional funding. At a minimum with two ICs, we plan to recruit 120 donors, as this is a minimum, not a maximum. We would like to emphasize this is not a standalone project, and as a pilot project, it has the potential to do more as additional ICs sign on. For $10 million per year total cost, we can recruit 200 donors, increasing our statistical power, thereby creating a valuable, deeper and comprehensive resource of gene expression protocols. There are some economies of scale here, while sampling costs are fixed, we expect other costs such as sequencing to decrease and add more donors as a result. I'd like to end with how DGTEX addresses key strategic needs. This is an opportunity to build upon functional genomics by establishing roles of genes and regulatory elements in developmental pathways and networks. And it also fills a gap in genomic data resources that lack data from early developmental stages. We do also have the potential to do work with model organisms for comparison and basis for downstream experimental studies. Thank you for your time and attention, and we will start the council discussion with Dr. Chung, Dr. Gravely, and Dr. Pritchard. Go ahead and start. So I think it's really important in terms of having kids involved in this. I know it's not something that necessarily NIH can address right now, but even more development is going on before birth. And at some point, that's a gap to be filled, whether it's with other model organisms or groups or whatever. But just realize that that's an important gap. Within this, I know you're addressing it just to emphasize it's a tough situation in terms of being able to collect these samples. Thankfully, healthy children don't die often. And so being able to get those situations is tough. I think you're taking the right strategies in terms of this. I guess within that, though, one of the things I'd emphasize in is if these are meant to be healthy, normal children, to make sure they really are healthy, normal children. Some of the individuals that die, we only find afterwards if we do a molecular autopsy that they weren't so normal. So depending on what age you're getting, but especially the newborns, at least my recommendation would be actually do a genome sequence first to make sure that they really are the healthy, normal individuals you think they are. And then the one other thing that I'll add is that the age groups that I think I saw pass by, you were putting them in pre-pubertyl or post-pubertyl by a particular age group. Number one, it doesn't, like biology, doesn't actually follow those ages so particularly. So just make sure you're classified by biological development rather than a strict age definition in terms of that. And in particular, the reason I'd bring up this distinction is that many of us think that, for instance, adult diseases may be very much dependent on what happens during development in puberty. And that pubertal status might be really important in terms of understanding adult determinants of health and how they are developed in childhood. And so just as a specific use case, breast cancer, for instance, may have a lot to do with what happens with the ductal cells during breast developments and mutations that are acquired during that period and other things. And so just getting that developmental stage is important. That's great. Thank you for that feedback. And just to reiterate, we will have apothologists review the tissue slides before incorporating them into the DG textbook. OK, Jonathan, on the phone, you want to go next? Sure. Yeah, I think that this is an important project. That's great. I was wondering if you could say a little bit more about bio-banking for tissues and how you imagine that the community is going to get access to that and what sort of quantities of tissues there will be, that sort of thing. So the quantity of tissue, as I remember from G-TEX, they collected a dime-size tissue sample. And as far as access for the tissue samples, it will be available for the network. I don't think we've really figured out the details for investigators outside the network. But I might ask my G-TEX colleague if there were requests for samples outside of the G-TEX network. Yes, there were requests. And we established a policy that was implemented to have outside people from the consortium to access the samples. And we received, I would say, not many, but probably between 20 to 30 requests over the years to access the samples, to do any type of work from methylation work, all that kind of things. So I think that it will be down to establish a policy, most likely similar to that of G-TEX. Cell lines were produced for G-TEX as well. Anything else, Jonathan? No, that's fine. Thank you. I think it's an important project. I think it's great. OK. Dr. Gravely, anything from you? Sure. Again, I want to express my support for this. I think this would be a great compliment to the G-TEX project itself. And I think the issues of getting the samples is obviously the trickiest one for this particular project. I just was curious about how you're going to decide how much of the project should be done as chunks of tissue versus single cell analysis. Can you rephrase your question again? I guess there's some. Single cell analysis versus your findings. Oh, OK. Well, there was just some discussion of using some single cell sequencing technologies. So how much of that would be done versus bulk tissue? Or would that be more up to the applicants to propose? So as a network, I would think we would decide which single cells analysis that we'd want to do. I think the important thing is to make sure that we can capture or we can collect the specimens for that and then decide later which one would undergo single cell analysis. Sure? Well, I guess I'll say it before Jeff does. I mean, this, I think, there's definitely going to be multiple LC components to this, particularly with regard to making all the data public and trying to explain this to parents because it's very different than the normal discussion around autopsies, which is often about trying to better understand why their child had the disease they did or what led to their death and things like that. So it's a very different discussion. And I would think incorporating some very clear LC components into this would be important. Thank you. There's no mention of gender or ethnicity or BMI or diversity of any type. Can you speak to that? Yeah, so obviously we can't tell ahead of time how many males, females, and ethnicity, but we would write it into the FOA to make sure that we do have a diverse donor participation. I was thankful that you said that the 120 is a basement, not a ceiling, because certainly the power and the number of the genes that are detected by GTex is in upon sample size. The one thing that I was wondering about, you also say 20 to 30 tissues. And in some ways, 20 to 30 tissues in this age range would be great to have the same tissues as in the adult GTex. But the adult GTex also omitted a lot of tissues that would be of interest. One obvious is by which has a big change in overdevelopmental time. So what's the plan for deciding what tissues and the number of tissues? Is that again a basement or a ceiling? And can you give us some insight on that? Sure. Yeah, so the selection of tissue will be dependent on what's accessible. Again, we want to be mindful of religious and cultural differences. So again, we would need to see what tissues we can collect at the time. And also, we would also look at the RIN number, so the RNA integrity index. And I believe for GTex, it was over five that they collected or they accepted the tissue sample. So we would try to use the same model. So just to clarify then, are you planning on restricting it to the same subset of tissues that were originally collected in GTex? Or are you also going to consider tissues that were not collected in GTex? We can consider tissues that were not collected in GTex. I don't know what the autopsy rate is for kids. But I'm wondering whether you thought about accessing primarily kids who are already undergoing autopsy. That would obviously give you a full anatomic report, and plus would give you access to tissues that I'm going to guess are going to be otherwise difficult, perhaps, to get permission for, like brain or heart or some of the ones that parents may be more sensitive about. Any thought about whether kids undergoing autopsy would be a good source of samples for this? That's a good question. I think we would have to work. Again, we're going to work with the Oregon Precurement organizations and the medical examiners to see what would be feasible. So, Mona, during GTex, I thought the finding was that conventional autopsies weren't quite good enough. You really had to have a much more rapid. We had what we called rapid autopsy. Right. Rapid autopsy. So we had some donor coming from that side. But the autopsy had to be done very quickly. So, Jeff, your question is you have the small fraction of cases where there's going to be an odd habit. You can't just sort of take a subset of those and get into that because it's going to have to be in a more rapid decision-making process, which can probably be a small subset of a small subset. But doable. Yeah. I mean, if they were available, we would certainly take them. Yeah. And we anticipate this is going to be challenging to recruit. But we see it as a worthwhile goal. We had the same concerns about the adult GTex. Again, I'll just emphasize because the number of cases that qualify, it's usually someone who's on a ventilator and you're withdrawing support to get the autopsy you need to, is the brain banks have been working on this. To my knowledge, though, they're not using the rest of the body. And so it's just if they've already agreed to donate the bank, it's very likely those same families will donate other body parts. So. Great. Thank you. Other questions? I've got a countdown assistant over here. Can I get a motion to approve the concept? Second. All in favor? Any opposed? Any abstentions? Thank you very much. Thank you to Jyoti.