 Okay, we're making good progress here. Can we continue right along now with the third concept? This is now for an RFA, and Tina Gatlin is going to give the presentation for TECDEV for single molecule protein sequencing. Tina, go ahead. Okay, thank you, Rudy. So this concept arose as part of the strategic planning process where a group of my colleagues were tasked by Eric to reevaluate NHGRI's role in the proteomics space and also to consider new opportunities. And after receiving input from the community and research, we decided to focus on expanding our role in a very targeted area of technology development, so single molecule protein sequencing, which we see as a promising new field which fits with our scientific mission. I'd like to start off this presentation with an acknowledgement and thank each of my other colleagues who helped to shape this initiative. So Valentina, Ajay and Adam, this was a team effort and they will be available for the discussion after their presentation, as well as Carolyn. So next slide. The purpose of this initiative is to accelerate innovation and development in the growing field of single molecule protein sequencing, or SNPS. The ultimate long-term goal is to achieve technological advances to the level where protein sequencing data can be used for genome-wide surveys. And with enough speed, sensitivity, quantitation and accuracy to be used routinely in studies of genome biology and function, and as well as biomedical research in general. Next, we will be taking what we learned from our long-standing and successful efforts in DNA sequencing and applied to the proteome at scale. We view this as an exploratory endeavor to determine feasibility, and we are proposing a modest budget to assess such feasibility. And this is considered to be a high-risk technology development challenge, and if successful in the long term, it could transform the use of proteomics in much the same way as modern next-generation DNA sequencing and transform genomics due to its high throughput, low cost and generalizability. Next slide. For some background, so we all know that compared to DNA with its four nucleic acid subunits, proteins with their 20 amino acid building blocks convey a much greater array of chemical diversity. Proteome makes the human proteome extremely complex, and to add to that complexity, a typical human cell expresses over 10,000 unique protein gene products and can contain 100 times as many proteal forms for each gene product. Those different forms of a protein include post-translational modifications, SNPs, and splice variants. The dynamic range of proteins within a human cell can range from 7 to 10 orders of magnitude. Next. Currently, there are two main approaches to measuring proteins, including affinity reagents and mass spectrometry. While these two approaches are extremely valuable, they have their limits with respect to proteome-wide detection. These reagents have excellent spatial resolution, can detect small amounts of protein, but they rely on custom reagents and are limited in scale as you have to know exactly what protein you are looking for. Mass spectrometry has long been the dominant technology for large-scale protein sequencing and quantitation. However, it lacks the dynamic range and sensitivity needed to routinely detect low-abundance proteins and is also limited in its ability to map post-translational modifications. Next. Thus, there currently exists no technologies for routine proteome-scale sequencing and quantitation, proteome-scale being driven mainly by the need for speed, sensitivity, and dynamic range. So next slide. So why are we putting forward this SNPS single molecule protein sequencing concept now? There have been some promising technological advances, including in the nanopore space where much of the development seems to be taking place, and also in the area of admin-like degradation chemistry. There are also now a handful of companies in the space doing early-stage development. Thus, with the level of activities and academics and industry, we believe now is an opportune time to help advance the state of the art. There has been no funding opportunity available to date to advance the specific field, and dedicated funding could lead to the realization of such technologies and would help facilitate the detection of low-abundance proteins and enable true single-cell protein analysis at high throughput, and also enable improved cataloging of protein gene products encoded in the human genome, along with advancing the search for the so-called missing proteins that have never been directly identified despite evidence of existence at the DNA or RNA level. So next slide. So why NHGRI? Why should NHGRI be investing in this area when our extramural investments in the proteomic space has been historically low? So first, one of the success stories of NHGRI, of course, as we all know, is our leadership in advancing genome technology development, and we would be taking the lessons learned in DNA sequencing technology and extending into the proteome world, especially with respect to scale, along with pushing towards quantitation and high dynamic range samples, and a very long-term goal would be accurate de novo sequencing versus relying on partial sequence information, although partial protein sequence does provide a tremendous body of knowledge. Next. So second, this technology has the potential to expand our understanding of genome biology and function, which is an important overarching theme in NHGRI's strategic planning process. There is recognition that data types beyond primary DNA sequence, including readouts from proteins and metabolites, are needed to fully understand genome function in different biological contexts. In this sense, we will eventually need protein information to fully understand how we go from genotype to phenotype. The high sensitivity nature of SMPS techniques has the potential to enable true single cell analysis where a lot of important genome biology takes place. Another important area is to establish roles of all protein coding genes in pathways and networks, which is, which this technology may help facilitate because of its potential to scale proteome-wide. And then lastly, better multi-ohmic molecular diagnostics and modeling are needed to improve disease prediction and prognosis. Generally low abundance of disease protein biomarkers, the high sensitivity nature of SMPS could add an important data type for disease diagnostics and modeling. So next slide. The scope and objectives for this initiative are to support investigator-initiated research with the aim to significantly advance single-molecular protein sequencing technologies. We want proposals to be novel, allowing for high risk, and we are not seeking proposals that are only putting forward incremental advances. Some examples of the types of techniques for single-molecular protein sequencing that we would consider to be appropriate for development for this initiative include nanopore sequencing, admin-like protein degradation coupled with massively parallel fluorescent measurements, other fluorescent-based techniques, recognition tunneling. And while the listed technologies are all utilizing amino acid sequencing approaches, we would be open to other technologies such as DNA readouts as a proxy for protein sequencing. Other technologies we may not know about yet or that may not even exist. The key is that the technologies have potential to scale genome-wide. So next, techniques that would not be appropriate include mass spectrometry. So there are plenty of other initiatives and ICs that cover the space well enough. And with our modest investment, we do need to bound the scope at some level and not leave wide open. And again, to emphasize another way, technologies that are not on the path scale would not be considered within scope. Next slide. For mechanism and budget, so similar to the genome technology development RFAs that were approved last September and recently released. So we are also proposing a mix of R01, R21, and SBIR funding mechanisms as RFAs. The plan duration of the program is five years as a first round of funding. And per the table, the program would ramp to $9 million a year by year three. And if after an evaluation in year three, it's decided that the program should continue and funds would be proposed to be maintained at $9 million a year. So commitment for the five years without any renewal is $29 million. And I will point out that that $29 million includes SBIR dollars. But as a reminder, small business funds come from a set aside. And so the total dollars without the SBIR would be $21 million for R01 and R21 grants. So due to the exploratory nature of the initiative, awards will be a little smaller with shorter duration than usual. So R01s would be up to 500k direct costs per year for up to three years. R21s up to 200k for two years. And then SBIRs, the total costs up to 250k for R43 phase one and then up to $2 million for R44 phase two. So we are seeking a sign on from other ICs. NHGRI is a small player in proteomics with our investment representing 1 to 2% of NIH investments. And while this technology will contribute to NHGRI scientific mission areas, successful development of this technology will be of course a benefit to the whole biomedical research enterprise. So we do hope that other ICs will participate. Next slide. So just to place this initiative in the context of NHGRI's overall technology development spending. So as a reminder to most of you, so this is a slide adapted from Mike Smith's September Council presentation where he presented that set of initiatives to expand technology development. So in FY19 investments were 29 million and supported by a mix of RFAs, PAs and PARs. And that 28 million is also the previous five year average of technology development investments. And then the September proposal was to grow to 45 million by FY23. So a 15% year growth over a five year time period. And these dollars are just for R1s and R21s and does not include SBIR funds. Next, our single molecule protein sequencing proposal as I mentioned on the last slide is to ramp to $9 million a year by FY23. However, subtracting out the SBIR dollars for direct comparison, this is $7 million a year. And so the protein sequencing investment could be viewed in this way as being about one seventh of the future total investment. And so we view this to be about proportional to our priorities in technology development. So as a last comment from NHGRI's experience with tech development efforts, we know that this will require a long term coordinated effort to achieve long term goals. And while it is tempting to think that single molecule protein sequencing is just too challenging of an endeavor, this is a time when even modest advances could have great utility. So an organized exploratory investment would help stimulate the field. And it's also an opportunity for NHGRI to try something new. Next slide. So now is the time for questions and discussion. And so we did have three council members who took a closer look at this concept so I'd like to call on them to lead off the discussion. So they are Dr. Steve Voter, Howard Chang and Olga Troyeskaya who hopefully is back online. So Steve, would you like to start us off first? Sure, Tina. Thank you and thanks for the nice presentation. I'm excited about this program. I think it's a very worthwhile program for the NHGRI. A couple of points that I'd just like to reiterate that you covered. First is that it's consistent with the strategic goal of filling in the molecular gaps between genotypes and phenotypes. And in order to do that, I think you've taken a very focused and targeted approach, as you mentioned, to insist that the technologies are going to be high resolution single molecule approaches. And importantly, that they have the potential to scale in a highly parallel format to complement the type of information that we can currently get from genome wide sequencing abilities. And this isn't just a free for all in technology development. This is going to be judged based on high sensitivity and also ability to scale to give a complimentary view of a whole genome or a pan genome view. As you mentioned, also new amino acids are particularly challenging. The chemistry is really challenging. However, there have been some potentially whole genome approaches that have been articulated lately. And part of what we maybe could hope to realize in the near term is, you know, ultimately we'd like to go to single amino acid by amino acid sequencing a basis, of course, that may not come easily. There are some very innovative ideas recently about how to do some genomic fingerprinting or excuse me proteomic fingerprinting by by looking at a limited number of amino acids and their spatial distributions in fragments of DNA. And of course these can be coupled with bioinformatic approaches based on predictions that we know about what should be in the genome. So we may realize some early wins in this and I hope that's true. And technologies that go all the way from a rebirth of Edmund degradation to adaptation of Nanopore approaches just to name a few. The final comment I'd like to make is that, as you pointed out, this will be a five year program, and I'm sure we'll be reviewing it periodically during that five year window. But this is a highly risky venture, which is good, because we need to inspire innovations in this area. It also inspires some new ideas about how to do genome wide assays for other molecules of the genome. And it'll be important for the coordinating center in the future to be evaluating these and to make timely reports back to us to measure success and to give feedback on further funding. So that's all I wanted to say but I'm very supportive of this program. Thank you, Steve. Howard or Olga. Hi, everyone. Nice to be here as a panelist actually sorry. I'm also very supportive of the program I think it's clearly a really critical area and something that we really need to do to be able to understand the genome and as such is clearly very relevant to NHGRI central mission. I do also agree with Steve and I expect Howard from our prior discussions in that we want to think it would be great if you guys think broadly about that specific types of novel technologies that would be considered responsive. With flip side I think it's really critical to figure out how to assess what really is considered properly properly scaled and protein wide and properly sensitive since of course these will be novel technologies and it's very easy to claim protein protein wide for anything so I think it's worth thinking ahead of time about some clear guidance on what assessments. The responses should provide and how they could really support the fact that these technologies are responsive to your goal of being protein wide and highly sensitive and reasonably accurate. And as such for many of them I would expect that associated computational analyses are likely to be critical. So it's not that you know that that this should be the central goal and completely computational methods would be responsive but for the cases where computational analysis would be really critical for making the technology appropriately accurate appropriately protein wide. I think it's important that they don't end up being an afterthought that is promised to be thought about next five years. It's a really great program I'm very supportive. Thank you. We'll definitely take those comments into consideration when it comes to FOA writing. Our. Yes. So Tina thank you for the presentation. I appreciate very much that you taken into the taken into comments from a subset of the, the council members and have sort of crafted this proposal. So I'm highly supportive of this idea. I agree that this is I think certainly in the scope of NHGRI's mission and point to understand the proteome and indeed this is a likely high impact and this is really a timely sort of proposal right now. So two sort of comments I have number one as Olga said, kind of the metrics for defining success, especially in the interim sort of your time for what would that look like. We know that we have the ultimate aim as Steve pointed out amino acid by amino acid, a single molecule protein sequencing, but obviously that's that's very much be asking for so what what is the interim success look like. What I think is about the coordination among these projects so this is was outlined as a R1 R21, not like an R1 or a U01 mechanism. So is expected that NIH was somehow coordinate and share coordinate communication between these different teams to reach this goal of a single molecule sequencing, or is it better other people have they're trying completely different ideas that they should go off on their own a bit. This is something that's up for discussion. I just want to close by saying that I think that now we witness kind of the phase of the explosion and improving the technology for DNA sequencing, and that led to many other sort of problems being turned into DNA sequencing problems that you can then solve them. And so it's possible that those kind of innovative thinking and technologies in genome science cannot be brought to this new frontier. Thank you. Thank you Howard Steve rich. Yeah, thank you. I'm supportive of this program in part. This is something that multiple institutes are interested in a lot of the work that's been performed out of NHLBI for example and their top med program which generated whole genome sequence RNA sequence DNA transmission, metabolomics and some proteomics data really opened up the question of just how good this proteomics data are. And, and I think, comparing different methods of generating proteomics data has shown that in some cases it's good, I correlated results and other cases very poor. And I think this initiative will help really set the stage for high quality understanding of the proteome with respect to the genome as well. And institutes at specifically NHLBI NCI, and in fact the American Heart Association precision cardiovascular medicine institute has been holding meetings for the last year or more about proteomics and how to move it forward. If you're looking for other ICs, then obviously the first place to start would be NHLBI and NCI and both are sort of awesome who was that NHLBI is now at NCI and very much part of pushing this forward so hopefully you can get people to help carry the weight. Yeah, we've been certainly in contact with our IC colleagues for quite some time, including NHLBI and knowing very well what their efforts are in top med. So, and so the other big ICs that have a lot of support for proteomics include NIGMS and NCI, and also NIAID. So, we're hoping to get one of those four or more participating. Great. Thank you. Sharon Pawn. Yeah, so I did just really want to encourage working with the other institutes and so NHGRI doesn't wind up shouldering the bulk of the funding because this really does apply across NIH. But I wanted to follow up on something Steve just said that, you know, there's a mention in the concept about having an annual meeting. It's great, especially if we're allowed to travel. But I do think maybe thinking more creatively about how the funded groups would really need to share, whether there's certain samples they might all analyze and routinely compare the results are really ways to make sure that people aren't just working in a silo on their method and making sure that they don't have to travel IP but really more crosstalk. It doesn't have to be a consortium, but more crosstalk so you, we really do get at the end of this methods where we can compare the results and get some ideas of the best use of each of the methods. Thanks Sharon. Yeah, and so we decided to model this program after really the current tech dev program, you know, which is very R based and they have an annual meeting where they share their data and, you know, it's an opportunity to collaborate and learn from each other. But yeah, thanks for, you know, considering to think about other ways to coordinate. Other comments from other council members. I'll just say I am very supportive. Thank you, Rafa. Okay, can I get a motion to approve the concept. In a second. Second. All in favor. I, I, anyone opposed. Any abstentions. Thank you all.