ProQR Therapeutics N.V. (PRQR) Earnings Call Transcript & Summary

Good morning, and welcome to the ProQR Therapeutics Analyst and Investor R&D event. [Operator Instructions] At this time, I would like to turn the call over to your host, Sarah Kiely, Vice President of Investor Relations and Corporate Affairs at ProQR Therapeutics. Please go ahead, Sarah.

Thank you, and good day, everyone. We appreciate you joining our event today. I'm Sarah Kiely, Vice President of Investor Relations and Corporate Affairs at ProQR . Today's event will take an in-depth look at our Axiomer RNA editing platform technology and our plans to advance it. On Slide 2, you'll find the agenda for the event and our speakers. From the management team are Daniel de Boer, our Founder and CEO; Gerard Platenburg, our Chief Scientific Officer; and Rene Beukema, our Chief Corporate Development Officer and General Counsel. We're also very pleased to have Dr. Peter Beal of UC Davis, present on ADAR. Following the presentation, we will have a management team and Q&A session for covering analysts before we conclude the event. Today's event is being recorded, and we will have a replay available on our website following the event. During the presentation today, we will make forward-looking statements there are risks and uncertainties associated with an investment in ProQR, which are described in detail in our SEC filings. I'll now turn the presentation over to Daniel. Daniel?

Thank you, Sarah, and good morning and good afternoon, everyone, and thank you for joining us today. We're excited to share with you today a comprehensive update on the company following the decision taken last summer to fully focus our strategy on advancing our proprietary Axiomer RNA editing platform technology. We believe this platform holds great promise to treat diseases that are otherwise untreatable. ProQR is exclusively focused on the development of our proprietary Axiomer RNA editing platform. which has brought applicability across multiple therapeutic areas. Our initial focus has been on CNS and liver and we will announce today our initial pipeline targets during the event. These targets will focus on diseases that originate in the liver. The Axiomer technology was invented at the ProQR Labs back in 2014 and uses the well-proven modality of oligonucleotides to recruit a novel mechanism of action. Axiomer uses an editing oligonucleotide or EON, to recruit and [indiscernible] ADAR, which can microsurgically edit the RNA, changing an A or an adenosine into an I or an Inosine which is subsequently read as if it was a [indiscernible] or G. ADAR is present in all human cells and RNA editing is a naturally occurring process. And in fact, it is happening in all of us right now as we sit here. Our Axiomer platform makes use of the ADAR machinery that nature has developed and recruits it to added specific adenosines in a very targeted way. Our strategy includes both in-house development of pipeline programs using this technology as well as selectively partnering on noncore targets, allowing us to capture the full value of this platform technology. And as we will also delve into today, preclinical platform data demonstrated that ProQR's Axiomer RNA editing technology is broadly validated across multiple genes. At ProQR, we believe that RNA editing will become an important pillar within the field of genetic medicines. Over the last years, we have seen the field of genetic medicines maturing and leading to more and more important medicines for patients. And RNA plays an important role in that. We've seen multiple approvals over the years of RNA medicines and of course, most recently, the mRNA COVID vaccines. Many large companies are making significant investments in genetic medicine, placing an important bet for the future on this class of medicines. And within that field of genetic medicine, we are convinced that RNA editing will become as impactful as, for example, siRNA by adding a novel endogenous tool to the arsenal. As the leader of the RNA editing field, we have spent the last 8-plus years since the discovery of the technology, optimizing the Axiomer platform in our ProQR labs and in partnership with academic partners to drive a robust understanding of ADAR biology and the design rules. We see RNA editing as the next evolution in the RNA medicine field. Similar to RNAi, we are recruiting an inductions mechanism that's already present in human cells. But instead of locking down an entire gene, we can now edit individual basis in the RNA. This opens the door to develop treatments for diseases that are otherwise not treatable. And in these instances, we have unique advantages over other technologies. Axiomer is a very versatile platform, allowing us to target a wide range of disease and can potentially lead to hundreds of medicines in very different disease types with a variety of molecular mechanisms. As we can make edits to the RNA, we don't have to touch the DNA or make any permanent changes, which comes with significant safety advantages. With transient editing, we can expect to dose quite infrequently, maybe 2 to 4 times a year. And as our editing is strengthened, it can also be applied in situations where permanent changes would be deleterious. Axiomer editing is done with high specificity, minimizing off-target effects, and we don't have to use viral factors. We can lean on well-established delivery routes and moieties that have been developed and proven over the last decades. We see Axiomer as an elegant platform using endogenous ADAR machinery that essentially leverages the body's own potential to treat disease. ProQR has quite literally led the field of RNA editing since 2014. And when procure scientists invented the RNA editing using induces ADAR and performed the first experiments using our editing oligonucleotides to recruit natural and [ inductively ] expressed ADARs. These experiments also led to our first IP filings for this technology back in 2014, which laid the foundation for our leading IP estates today. Over the subsequent years, we have done considerable work to optimize and enable the development of highly specific, safe and efficacious platform. And today, we will share extensive data demonstrating its capable of efficient in vivo editing for therapeutic uses. We plan to capture the value of our platform across 2 key strategies: first, through the development of an internal pipeline of high-impact medicines and second, through selective partnering. As we will announce today, ProQR will initially focus on translating the Axiomer technology into the clinic in 2 separate development programs that can yield potentially very important therapies. The initial programs also bear the burden of validating the broader Axiomer platform technology in humans. Our pipeline will focus initially on the delivery of our editing oligonucleotides to liver as delivery to that organ is largely derisked, and we can build on proven delivery technology to the liver. For the avoidance of doubt, this does not mean that we will focus on liver disease only. Many diseases are caused by proteins that are expressed in the liver. And by targeting the liver, we can treat diseases throughout the entire human body as we will present to you today. As we will advance these first 2 and subsequently more programs into the clinic, we plan to select some of these programs for developments to markets by ourselves. And for large indications, we plan to advance these through clinical proof of concept and then seek to partner this out for continued development. This speaks to the second pillar of our strategy, partnering. Partnerships are a great way to advance applications of the platform beyond the targets ProQR intends to develop independently. In addition, these partnerships bring important validation. External [ eyes ] in the science and nondiluted funding to help us fund our operations. We will selectively enter into partnerships that help us advance the platform and the business. For example, we have partnered with Eli Lilly on concurrently 10 targets on the Axiomer platform technology, where ProQR leads to the discovery phase and Lilly leads all phases beyond that. This partnership so far brought in $125 million in upfront payments and we expect to receive another $50 million from Lilly likely this calendar year for an option that they hold to expand the partnership to 15 targets in total. In addition to this, ProQR is eligible to receive $3.75 billion in milestone payments, including discovery, nonclinical early clinical milestones plus royalties. We're very pleased with this partnership and with Eli Lilly as a partner, and we continue to execute on this partnership with high priority. Given the vast opportunity with the platform, we have appetite and capacity to selectively form additional multi-target discovery partnerships. As we advance our pipeline, our R&D specially takes a risk-mitigated approach in a translation into the clinic to maximize the probability of success in this space. It's worth to acknowledge the learnings we have had in the previous 10 years in translating multiple therapeutics from the lab into the clinic as those equip us for success in the future. For example, we have selected targets that are deep routing in human genetics, where we know that the post-editing RNA products is associated with beneficial impact on human health. We prioritize targets that are expressed in liver as liver delivery of oligonucleotides to hepatocytes being derisked, allowing us to study this new mechanism of action in isolation of other variables. And for the initial targets in our in-house pipeline for programs that are based on Axiomer, we have selected targets that have strong translational tools available, including animal models that are representative of the clinical situation and clinical biomarkers that allow to get objective confirmation of target engagement via [ Blotra ] to gain solid validating platform data early in the platform development. Incorporating all of our translational and drug development experience, we are confident that this risk mitigation mitigating translational strategy we can study the new mechanism of action of RNA editing with high probability of success into the clinic. Today, we are announcing our internal initial pipeline programs, and [indiscernible] will go through this in quite some detail later in today's program. On this slide, we are displaying our pipeline. As you will see, our pipeline contains the mix of targets for rare and prevalent diseases as well as wholly owned and partnered programs. We will initially prioritize the following 2 programs to move forward into the clinic. Programs that we believe carry all the characteristics to translate the platform into the clinic with high probability of success, including a solid routing in human genetics. Firstly, we will develop AX-810 for cholestatic diseases, targeting the NTCP channel. And secondly, we will advance AX-1412 into development, which will be developed for cardiovascular disease starting the B4GLT1G. With AX-1412, our intention is to develop this to an early clinical proof of concept and then partner for further development, given this is a large population indication. Gerard will detail these programs later on into the event. And in addition to these programs, you will find a range of programs for other indications, including NASH and other cardiovascular targets metabolic disease targets, neural disease targets and also other rare diseases. All programs that at some point in the future, we will provide and AX-810 and AX-1412. Although the RNA editing field is early, ProQR is well positioned to execute on the Axiomer business plan. Back in 2014, we invented this technology. And over the years, we've been able to really focus on the basic science underlying the ADAR biology and optimizing the design rules for the Axiomer platform. As we'll describe today, there are a large number of potential therapeutic applications in both prevalent and rare diseases. We will also present a range of preclinical proof-of-concept data, including in vitro and in vivo studies, involving organizes, mice, nonhuman primate experiments in liver and CNS. For better or worse, broker has gained quite a bit of translational and development experience in oligonucleotides over the last decade. We can integrate all of our lessons learned into translating the Axiomer platform into medicines for patients. In that same vein, we have a lot of experience with target hunting and [indiscernible] We have, therefore, selected targets for our pipeline that both have high probability of technical success and could lead to important products with viable business cases. Axiomer is protected by more than 10 patent families, that have multiple granted claims, protecting the recruitment of inductions ADAR with editing oligonucotides in a very broad way, dating back to the invention in 2014. These patents have been opposed and survived that proposition. Hence, we feel very confident about the strength of our leading IP estate in the field of inductions ADAR editing with editing oligonucotides. Our partnership with Lilly provides us with strong validation, a great partner to advance potentially important medicines. And is providing a source of nondilutive funding to help us fund our operations. Beyond our existing partnership with Eli Lilly, we plan to selectively enter into additional multi-target discovery partnerships. We have indebted our team, management board and advisers for the Axiomer business plan. This includes John Maraganore, joining our Board as a strategic adviser back in 2022. John built on Alnylam a successful company over the last 2 decades, and we can leverage his experience to apply Alnylam playbook to build out the Axiomer platform. Our Scientific Advisory Board has great expertise in RNA, including Phil Zamore and inventor of the Nobel prize-winning RNAi technology, Martin Maier from Alnylam, [indiscernible] from avidity and Peter Beal, one of the leading ADAR experts in the field who will present today as well. James Shannon, the former Chief Medical Officer at Novartis; and Yi-Tao Yu, Professor at Rochester and experts in the field of RNA editing complete our Scientific Advisory Board. And last but not least, ProQR is well funded with a runway into mid-2026, in which time frame we anticipate having clinical data from multiple development programs. Against this backdrop,We're very pleased to now hand the call over to Gerard Platenburg, who will guide us on a deep dive through ADAR Science, highlight our platform and a wide range of preclinical proof concept data and present our pipeline. Gerard, over to you.

Thank you, Daniel. It's my pleasure to introduce our RNA-based editing technology Axiomer. Axiomer, as you can see on the left of the slide, uses endogenous ADAR or adenosine [indiscernible] acting on RNA to convert adenosine-inosine, a process which takes place in every cell in our body with different functions as you will learn. We will start video on ADAR, followed by a presentation by Dr. Beal, who is a world-known experts on ADAR. And now we will start the video. Enjoy. [Presentation]

Now that you know more about the general information about ADAR, we are very pleased to have Dr. Beal today who will give us much more details about ADAR science and learnings in the field. Dr. Beal is a Professor and Department of Chemistry at the University of California at Davis and a world-known expert in ADAR science.

Thank the organizers for the opportunity to present our work on RNA editing by the ADAR enzymes and discuss our collaborative efforts with ProQR. So what I want to do today is give you some background information about RNA editing, ADAR RNA editing and the ADAR enzymes. And then talk about how we are using structural studies coupled with biochemical analysis to optimize the guiding strands for the ADAR enzymes for therapeutic editing work that we're doing in collaboration with ProQR. So before I start, though, on that, I want to talk a little bit about ADAR RNA editing. And so the reaction we're talking about is this reaction shown on this slide. This is the denomination of adenosine in an RNA molecule, and it can be in an mRNA and the conversion of adenosine via deamination reaction in mRNA generates in a [ c ], a minor RNA base. And because of its structural similarity to Guanosine, Inosine is actually translated as if they were Guanosine. So if this A is in the proper location and a [indiscernible], it can change the meaning of the [indiscernible]. And in this way, it edits the information in the RNA. And ADARs are human proteins. These enzymes that does reaction are human protein. So there are many known human substrates for the ADAR enzymes. And what I'm showing you now on the bottom of this slide is 1 of my favorite human substrates for the ADAR enzymes. This is the pre-mRNA for encoding the protein [indiscernible], okay? And at the junction between Intron 5 and exon 6, the RNA folds into a structure that is extensively duplex and it has an A at the specific location indicated by the red letter. This A is in an AAA codon for [ Lysine ]. And when the A -- the red A is deaminated by the editing enzyme, it generates A inosine A, which is a codon for [indiscernible]. So the [indiscernible] protein actually comes in 2 forms. It comes in the genomically encoded form that has lysine, but it also comes in the edited form that has arginine. And we published a paper quite a while ago now showing the -- this is in collaboration with my colleague, Sheila David, who is an expert on this [ NE-1 ] protein, it's a DNA repair enzyme. We showed that the 2 forms of the protein are biochemically distinct. So this is an example of how this ADAR enzyme, the enzyme that does the reaction is able to tune the properties of a protein by changing the amino acid in the protein. We now know that the side chain of the amino acid at position 242 [indiscernible] is in direct contact with the substrate for this enzyme in that way, it's tuning the properties of that protein. Now the enzymes that do this reaction, I've already introduced them as ADARs for adenosine deaminase that act on RNA. There are 3 human genes that encode ADARs, the ADAR1, ADAR2 and ADAR3 genes. There are 2 forms of ADAR1, the long form p150 and the short form p110. ADAR3 is not a functional deaminase. So it actually has a defective deaminase domain that is not capable of carrying out the reaction, but it is capable of binding RNA substrates. ADAR1 and 2 are functional the deaminase, right? And these are the purchases that we've been focused on in our lab. These enzymes can bind duplex RNA, and that's at least partially explained by the presence of what I referred to as dsRBDs or double-stranded RNA binding domain that are part of the protein structure. ADARs -- ADAR 1 has 3 dsRBDs, ADAR 2 has 2 dsRBDs. Now each of the ADAR also have a C terminal deaminase domain where the reaction takes place. And our lab has actually shown that the deaminase domain is also a duplex RNA-dependent component. So it's important to think about in the context of directed RNA editing, why we're here today, right, that the ADARs required duplex structure, okay? Duplex is essential for the substrate. Now the reaction of adenosine to inosine conversion and duplex RNA has multiple consequences. I've already talked about the recoding effect where if you have that A in a specific code one, domination can convert that code on to something else. And one of the more well-studied substrates for human ADAR2 is a glutamate receptor, which is a calcium ion channel, and it's known that the 2 forms of the protein have different ion conductors through the channel. It's also true that in the case of the secondary structure substrate for ADARs, [indiscernible] is often in an AU pair. And so the denomination of that creates an inosine-U mismatch, which destabilizes the secondary structure. And the reaction of double [indiscernible] RNA in the cytoplasm and weakening the secondary structure and preventing that RNA from binding to cytoplasmic receptors for duplex RNAs is one of the -- probably the most important biological function for human ADAR1. Preventing an immune response because those receptors are there for recognizing viral RNA. And there are other consequences of ADAR editing as well. ADAR editing can affect splicing. The reaction within microRNA precursors or micron binding sites also can affect expression of specific target genes. Now one thing that's gotten right, the community very excited and again, why we're here today is this idea that you can do directive editing, right? We're guiding oligonucleotide. Recall, I said that the requirement for ADAR recognition is that the RNA is in a duplex structure. But we can create that duplex with the synthetic oligonucleotide, right, binding to a particular transcript, which will then recruit the ADAR to do a reaction at a specific site. You can imagine we may want to convert an A that is there because of a G-to-A mutation that causes some genetic disease perhaps by creating a premature termination codon, right? We can recruit the ADAR to that site, convert that A to inosine which will now be translated as if it were a which could allow for read-through or some other therapeutic effect, repairing the defect or making a corrective edit, okay? So the guiding [indiscernible] Cytidine then would be the therapeutic industries. Now on this slide, I show you some key advances that key advances in ADAR research over [ rest ] of the last 30 years. Starting with the discovery of ADAR1 in 1988. Then here we have in 1995, the first report of directed RNA editing. [ Todd Wolfslab ] showed in Zenith [indiscernible], it could direct the ADAR reaction to make a corrective edit. In 1996, we see the discovery of ADAR2, right? And then in 2000, we see the role of ADAR2 and neurobiology groups. Recall, I indicated the importance of that glutamate receptor editing, that's a very important role to controlling the property of neurotransmitter receptors. We see in 2012, now others doing directed RNA editing with ADAR fusion proteins. Now it's not recruiting endogenous ate, this is actually generating a fusion protein where the 8-deaminase domain is connected to some other targeting element and directing it that way. In 2012, we see the role of ADAR1 established in the innate immunity, okay? And then as early as 2014, ProQR has shown, right, that you can do directed editing with endogenous ADARs. Because this is a very important advance, right? The idea that we could recruit endogenous ADAR to do a therapeutic edit. In 2016, our lab actually reported the crystal structures of human ADAR2 RNA complexes. Now this is important for the field because now this allows for rational design, rational optimization of the strand that's going to be the guiding strand for ADAR, the therapeutic and the direct-[indiscernible] application. Now in 2019, the field was really invigorated by the discovery of a role for ADAR1 in cancer. It's now known that certain cancers are addicted to ADAR's 1 activity and loss of ADAR activity is particularly lethal to those cancers. So ADAR1 is now a target for cancer chemotherapeutic development. And then in 2022, we see directed editing in primates with short time. So I believe you're going to hear more information from ProQR in this event about their work directing editing in primates with chemically synthesized short polygenic times. Now this idea of recruiting a human effector protein to all properties of RNA as outlined in this part of the slide here, where we could have an [indiscernible] Oligonucleotides, recruiting an ADAR to do corrective edit. I see this as very analogous to 2 other pathways that use synthetic Oligonucleotides to recruit human proteins to effect a change in RNA. Of course, I'm talking about antisense, where the use of ASOs and the antisense pathway will recruit RNA [indiscernible] to cleave particular target mRNAs causing a loss of function of that for that target. Also ran interference pathway, where an siRNAs guide strand recruits the human Ago2 protein to directed RNA for cleavage and loss of function, okay? So each of these pathways now have multiple FDA approved therapeutics. And I see this idea and this pathway for directed RNA editing in a similar vein and excited about the opportunity to participate in the any advancement of these [indiscernible] to the point where we can show that there's approved therapeutics in this pathway as well. Now importantly, the antisense and RNAi pathways lead to a loss of function, right? We're cleaving targets. We're losing function. But an exciting application of directed ADAR editing is we could have a gain of function, okay? So for instance, I made reference to this repair of a premature termination codon, right, we could actually restore the activity of a protein in a cellular cone that activity is lost with directed editing, which I think is a very exciting opportunity in this space of Oligonucleotides therapies. Okay. So if you're thinking about direct Readiness natural tenancy is to compare this to genome editing. And I think there are some interesting advantages to using directed RNA editing. But of course, the effects are reversible in the case of RNA editing, whereas in the case of genome editing, right, that's going to be a permanent change to the genome. In the case of RNA editing, there will be no need to deliver the enzyme because we're going to use ADAR as the effect for protein, right? It's endogenously expressed in human cells, whereas in the case of genome editing, one needs to deliver the cast protein. Of course, ADAR is a human protein and not one of bacterial origin and of course, we're not going to have the immune responses that we would have with a bacterial protein challenges do exist for the use of RNA editing. We would expect that we would have to have a continuos administration for the therapeutic effect. Whereas in the case of genome editing, it's likely would be a one-and-done sort of therapeutic. And for certain sequences, we see low editing efficiencies for directed RNA editing, okay? Now I see this actually as an opportunity for innovation, okay? So what we are very interested in doing and we are working with ProQR to do this to optimize the guiding Oligonucleotides to improve editing efficiency and using structure and biochemistry to guide that optimization process. And so for the rest of my presentation to a want to talk to you about some examples of where we've been doing that with ProQR. Okay. So in 2016, our lab working with my colleague here at UC Davis, Andy Fisher, solves the first crystal structure right, of an ADAR deaminase domain bound to RNA at a relevant point in the reaction pathway. So we did that by using a nucleoside analog that we had developed that mimics the deamination transition state, the adenosine deamination transition state. So we can trap this complex at a point along the reaction pathway that was catalyst be relevant. And then by solving the crystal structures then we could identify the key contact between the ADAR protein and the strands of the double vehicle substrate. And so we see multiple contacts to the strand that is edited, the strand that I have indicated in red here, but we also see multiple contacts to the strand that I've indicated in the blue here, right? This is the guiding stand. This is the EOR, right? This would be our therapeutic ultimately. And so we are now using the structural information, right, to optimize the individual components of this guiding strength, right? So I'm going to talk about how we're doing that. Now of course, we think about this guide strand right, as the drug, right? And we're trying to optimize the components of this guide strand. And I'm going to draw your attention to this nucleotide I've indicated as the orphan, right? This nucleotide right here is do nucleotide that was based paired to the reactive A. But in the process of the reaction, that flips out of the [indiscernible] and occupies the active side of the enzyme. After that base slipping step, this orphan position is left alone, we refer to it as the orphan position. I'm going to talk about how we've optimized that orphan nucleotide to maximize the amount of editing that we can see, okay? And I'm also going to talk about how we've optimized the minus 1 nucleotide position, right, for a sequence that's been a challenging sequence to target by AI using ADARs. All right. So let's start with the orphan position. So again, we have multiple crystal structures of the dams to man of at or 2 bound to RNA, right? And what I'm showing you here on the right-hand side of the slide is the Orphan C in contact to glutamate 488. This is the residue that's found in the endogenous ADAR, the wild-type enzyme Okay. We also saw the crystal structure of a hyperactive mutant of ADAR, okay, that has a glutamine at that position. And we see a very similar contact, okay? Glutamine, of course, can be a hydrogen bond donor and make a hydrogen bond to this nitrogen at the orphan position. Glutamate would be a hydrogen bond acceptor. And for that to actually make the same contact that glutamate would have to be protonated to be a glutamic acid and that's shown on this slide. So in this case, if you lose a proton, right, with proton right here, this would be a mismatched hydrogen bonding pair, okay? And with the glutamine, the glutamine always has a hydrogen bond donor, that would be a match pair hydrogen bond acceptor hydrogen bond donor. So this suggested to us, of course, because this is a hyperactive mutant that if we had an analog of the C that could donate a hydrogen bond, right, that could interact with the glutamate form of the protein, we might mimic the effect of the hyperactive mutants, okay? So this led us then. First, I should say, we tested this idea of a proteination-dependent interaction by studying the enzyme kinetics of the wild-type protein and the mutant at different pHs those studies supported this hypothesis that for the wild-type protein, there was a proteination-dependent effect consistent with this idea that you needed to have this hydrogen bond donor at this position. Okay? So this led us then. So this combination of crystal structure and biochemical analysis that led us to explore analogs for the orphan position. that had a hydrogen bond donor at this position, okay? And other ways was like normal [indiscernible] okay? And this led us to the testing of this analog that's referred to in the literature as benners based because Steven Benner developed this analog in his work with orthogonal base pairing systems for DNA, okay? So this was in the literature, but not for this application, right? We're using it in a new way here, okay? It has it is a cytidine analog with an N3 hydrogen bond donor. Now we showed in data that I'm not showing you here, the -- using enzyme kinetic analysis that base accelerated the rate of the reaction for both ADAR1 and ADAR2 in vitro. And so what I'm showing you on this slide now, is the use of that Z base in collaboration now with scientists at ProQR. This is an important collaborative effort here where our lab is doing the structural work and the biochemical analysis, and they're taking this information and they're using it and targets that are of interest to them. And what I'm showing you here is -- 2 different Oligonucleotides that are chemically modified and stabilized for metastability so improved metabolic stability. And the orphan position is varied to either a normal C or the Z base. And we see for 2 different targets, 1 target in mouse liver fibroblasts. Another target in retinal pigment epithelial cells. These are human cells as mouse cells. We see that the Z based modification, this 1 nucleotide modification at the orphan physician causes leads to a substantial improvement in the amount of editing that is seen. About 2 to threefold enhancement, and we know that our other targets lead to even greater enhancement of editing that is observed. So this 1 nucleotide is improving the editing yields, okay? So I want to just emphasize here the discovery of this nucleoside analog was enabled by the structural analysis, identifying the contact the biochemical analysis, exploring the nature of that contact and a chemical intuition about what it was needed to mimic the effect of a hyperactive mutant that then led us to this, okay? And this is ongoing work. This approach, I should say, is continuing in our collaboration with ProQR. Okay. So I want to tell you one other short story and a similar sort of idea, okay? So Here, what we're going to try to do is we're going to try to overcome the natural tendency of ADARs to react at sides that have certain nucleotides nearby poorly, okay? So it's known that have nearest neighbor preferences. It's well known and the literature that both ADAR1 and ADAR2 prefer to react at A that have either a U or an A on the fire prime side. And the natural substrates for these enzymes are depleted for those sequences that have either a C or particularly a G on the prime side. But there are therapeutically relevant target sides. For instance, premature termination codon that we may want to repair, right, that have G on the 5 prime side. So how are we going to enable editing at those, what would be naturally not preferred sides. So I'll tell you about that story. Now our structural studies explain this nearest neighbor preference because if we take -- if we look at our crystal structures, where the optimal 5 prime nearest neighbor sequence has been studied that has a 5 prime U and a pair with A. There is a loop of the prudent that has a glycine residue position 489 that comes very close to the minor group edge of that base pair. Now if we have a 5G the sequence that's not preferred by the enzyme, and we model a G-C pair at this position, right, mimicking what we see for 5G we see that there would be a clash with the protein at that position where the 2 minor group is in the minor group, okay? So how can we potentially overcome this? Well, very talented graduate student in my lab, Aaron Dougherty, who is now a post-doc with Jennifer Doudna at Berkley. What she did when she was in my lab is she actually explored different base pairing partners opposite that PMG for both ADAR2 and ADAR1 P110 and found that for ARI, there was clearly a preference to have a G opposite to 5 prime G. And for ADAR1, both A and G enhanced editing, okay, clearly better than the G-C pair that's shown here in green. So there's something going on with the puring mismatches, if you will, okay? And this was also shown in a different substrate for ADAR2 like the G-G pair seemed to be beneficial. So what's going on there? Well, we turn to, again, to crystallography, right? We generated a substrate that had a G-G pair next to an editing site, solve the crystal structure and found that G-G pair was in a very specific hydrogen bonding paired geometry, okay? The 5 prime G, the G of the substrate strand is now no longer in its normal anti confirmation, but it has flipped into what's referred to as a SIM confirmation and it's presenting what's referred to, it's hostage to the base in the guiding strand which is a G to form a specific pairing partner, okay? So why is that? How does that help? Well, that takes that amino group that was classing in the minor grew, right, and puts it into the major group. And that's actually shown, I think, more clearly in this bottom chemical structure formula here, where you see the 2 amino group and the minor group and a normal G-C pair when you have a G-SYNG-antiparing interaction that 2-amino group is moved into the major group and no longer clashes. However, we still have a 2 amino group from the G that is in our guiding strand. And we see that it's very close to G489, okay? Is there a way that we can potentially overcome that effect or improve it even further, I would say, Okay? So I told you about this GAnti-GSynpair, but recall that for ADAR1, the A also enhanced editing, right? So what could be going on when there's an A paired with G. Well, it turns out that there are structures of RNAs that have A-G pairs and the A and the G is in the SYN confirmation, but the A is protonated to form the same sort of pairing interaction. Now the pKA of A is around 3.7. So proteination at that site, right, will be unfavorable at physiological pH. But if you dig into the literature for nucleoside analogs, it's well established that 3-deaza and has a pKA of around 6.8. It's much easier to prudent that position than a physiological pH. And so this suggested to us that if we use 3-deazaA, we may be able to form the same sort of paring interaction, and potentially be able to improve editing even further, okay? And so we see that, in fact, when we compare Aanti to 3-deazaA, this is in vitro kinetic analysis, this is with ADAR1. We see an improvement in the catalytic rate of editing when we have 3-deazaA paired with G, okay? And we then went and solved the crystal structure of 3-deazaA paired with the 5 prime G, and it pairs as we had predicted, right, where the 5 Prime G is an Ascend confirmation, the 3-deazaA is base paired with that and a very nice pairing interaction, and we also see additional stabilization but the hydro bonded to its own phosphate. And importantly, when we overlay now the structure with 3-deazaA paired with G to the optimal 5 prime nearest neighbor, the 1 that has the 5 prime, we see nearly perfect overlap, okay? So using 3-deazaA to pair with G has created a structure that now to ADAR looks almost like the optimal 5 prime nearest neighbor, and we see this in improved rates of deamination Okay. So again, what we've done there is we've used the structural studies, right, to generate hypotheses about how the guiding strength could be optimized by using chemical changes to the structure of the components. -- of the guidance trend. And this is -- this, I think, is an exciting opportunity to continue to develop these guide strands for directed RNA editing. And we're continuing to do this in collaboration with ProQR. Okay. So to summarize, I told you that ADAR-mediated RNA editing is capable of rewriting genetic information, importantly, at the RNA level in a reversible manner, Human ADARs have important roles in neurobiology and innate immunity and cancer, okay? Synthetic Oligonucleotides. We refer to those as EON or editing Oligonucleotides, can be used some direct ADARs to make corrective edits. And we're using high-resolution structures of a RNA complexes along with rigorous biochemical analysis, we're actually measuring rates of deamination with purified proteins and substrates of varying structure and sequence. This has an enabled optimization of EONS by rational design, and I showed you 2 examples of that where we work together with ProQR Z at the orphan position and then 3-deazaA is at the minus 1 position, okay? And we continue to do this at other positions along the guiding strand. Different positions along guiding strand and also for different target segments. Okay. So I do want to acknowledge the people in my lab to do this work. I have a fantastic group of graduates and postdocs here at UC Davis have benefited from collaborations around the world, including with scientists at ProQR and I want to acknowledge the funding sources for work that we do on ADAR. And thank you again for the opportunity to present this work.

Thanks, Dr. Beal for a really excellent overall on our Axiomer platform, including some of the data that we've generated over the last months. And this section does contain data generated in-house, but also in collaboration, explaining the undisclosed targets in some of the following slides. As mentioned by Peter during his presentation and on the video you watched earlier, ADAR enzymes, Act on duplex RNA, so referenced as double-stranded RNAs. By learning from this natural process, we have developed our editing oligonucleotides EONs with the objective to mimic that. So we look very carefully what nature has given us editing oligonucleotides consists of short single-stranded RNA and I will get back to that a little bit later, that are actually complementary to target RNA. And the target RNA is indeed also a single-stranded molecule. And by binding it the editing oligonucleotides is create a double standard structure, which is resembling what you find in nature and then that will attract ADARs and allow the ADAR [indiscernible] editing to be performed. Our editing oligonucleotide are optimized to obtain very precise editing on target RNA, and I'm going to show you how in the next few slides. On this slide, you can see that the EON sequence defines the target RNA binding and provide stability. Our molecules are around 25 to 30 nucleotides and this size makes the molecules develop entity, as [indiscernible] speak. The 2 specific regions we can recognize in the yarns are described as follows. Firstly, we have to ADAR binding domain or ABR and that backbone modifications enable at binding and actually disable off-target editing. The second region we have is the editing-enabling region, ADAR and that actually is a very short region of about 3 nucleotides. And this is where ADAR is to achieve very specific A to I editing. This reading actually is designed to push the target into the catalytic domain of the enzyme and allowing the enzyme to perform its task. Over the years, we've developed our expertise and I very much understand how it's needed and what is needed to optimize the sequences that we are targeting. On this slide, you can actually see the advancements that was made over the last 8 years. and trying to understand and delineate the ground tools that are needed to design these molecules. Together with Dr. Beal at UC-Davis, we worked hard to understand the specific components that are needed and that are allowed by ADAR to be put into these molecules and to optimize the performance of such molecules. We changed the basis, the [indiscernible] modifications and the linkages, as you can see over here, and put in a lot of modifications in order to improve the pharmacodynamics, but also the kinetics of the molecules that we are using right now. This work led to our deep knowledge of the molecules that we are now designing and using for our development programs, but also led to a portfolio of our 10 foundational patent families, as you can see over here. In short, in my presentation, I will actually show that we have consistent RNA editing over all the models that we've tested in the nervous system and liver including none primary studies in vivo. I can show that we have used [indiscernible] to actually target the ERs to the liver, specifically to the hepatocytes which leads to increased editing, and that will be used in our subsequent development as well and by showing different applications of the technology. I'm going to show you the broad applicability of our Axiomer platform as well. In order to establish a strong platform, have established multiple cell systems and test systems in which we can test the performance of oligonucleotides. And we have been focusing to set the system up in the nervous system and the liver originated cells. And as such, we've developed different cell models, more complex models like organoids and then the in vivo models as well. The sum models as such, we can use optimize the performance, the organized systems, which are more complex, we are going to be using to actually test the effect of such variance in the excellent situation of the disease. And then we are going to be using the mice or none privates to study the performance in vivo. And uptake, effectivity and safety will be part of those studies as well. So let's dive into the CNS first. And here, you can see that looking at the neural models of neuronal origin. We've tested several of these EONs for their performance of editing. And you can see on the left, we bite you no uptake tested a EON targeting acting B. And you can see that we have a nice editing efficacy of over 50%. We took those learnings and we targeted a specific gene that expressed in roller cells, APP or amyloid precursor protein, and we were able to get over 20% editing as well. And it actually shows that in these 2 systems, or in the neuronal cells, the editing machinery is present and available and capable of performing the editing reaction. Next, we looked into more complex molecule systems to serve organoids, and that's shown on the next slide over here. Here, you can actually see that we were able to create and culture 3-dimensional structures containing a lot of different cells of the brain and mimicking thereby the real conditions and cell-to-cell interactions. In fact, you can actually distinguish certain cells and from the brain regions that they were derived from. So the experiment actually calls for a genetic uptake as we've done with cultured cells. And what you can actually see on the right panel is that we were able to get very high level of editing using [indiscernible], but also the [indiscernible] gave us 65% editing in the [ cebraorginoids ]. So this allows us to study the processes in, let's say, ex vivo circumstances much better. With that knowledge, we started to study the reaction in vivo and then we scaled up to vivo models at first in mice. this experiment in mice, we were able to dose these animals with a single dose through ICV injection. And after 4 weeks, we were able to show that we have extremely high editing in certain brain regions of up to 40%. But also, moreover, we could show that this 40% editing resulted in a 26 fold change in protein functionality which is actually a very nice proof that we can actually correct the G-to-A mutation. Following the results in the mice, we took the study into the nonhuman primates as can see over here. And once we've done is we dosed these animals with an intrathecal injection, and we tested the editing after 7 days using ddPCR. And as you can see, on the right we have in this fine region up to 50% editing as you can see over here. And if you analyze the brains of these animals, we can actually see up to 30% editing, which is extremely promising for applications in the nervous system as we move forward. So in summary, I think we've shown consistent editing of around 50% in all the cell models organoid models and the vivo studies that we found, which has a huge impact on our way of thinking of using this technology and moving forward into the development. As we've indicated before, we are going to be focusing on the liver for our internal pipeline development. And being at the origin of multiple diseases, the liver is really a target organ with a high potential to target with these editing therapies. So in these following slides, I will take you through some of the data that we've generated in the different cell models, organoids in vivo as we've done for the nervous system approach as well. We selected several cell models for the first step in the selection and the optimization of the EONs but mainly focusing on primary human hepatocytes as you can see over here. The left, you can see the experiment where we took the housekeeping gene acting B and incubated bigger cells. And you can see that we have over 70% editing, and that's something that really shows that the editing machinery is really present in the hepatocytes and able to perform the reaction. We took those learnings into a cell line that harvest the mutation that causes alpha-1 antitrypsin, as you can see on the right. And we were able to generate an EON, which resulted in very robust more than 50% editing. So that allows you to actually translate the learnings from the housekeeping gene into a clinically relevant target. And you can see that we have in both situations high level editing obtained. As the next step, we turn to liver microtissues because those are more complex organoids that allow us to show effect of certain treatments in more relevant models and you can see that the LMTs, as we call them, are coculture primary hepatocytes Kupffer cells and liver endothelial cells in a 3-dimensional form, which is then is forming a spheroid. On the left, you see a very nice live image of an LMT stained and you can actually see some of the more complex structures starting to form after a certain point in time, like the presence of bile channels using specific stainings. And that actually shows you and we'll use that in our later development program that certain essential processes can be mimicked in these very complex cultures, as you can see over here. We tested the editing potential of such LMTs on the right and incubating them with the housekeeping gene for [indiscernible]. We are showing very nice editing that increases over time showing the robustness of editing up to 40%, but also showing the endurance and the duration of the reaction as well. So this model allow us to really study processes that are important for product development later down the line. Having learned all of this, we took our lessons and then created data in vivo using wild-type mice, as you can see over here. In this particular case, we dosed these animal subcu with a 5x 10 mg per kg dose and tested the editing performance at day 7 after the last dose. And as you can see here, in wild-type mice, so no additional ADAR expression, we find over 40% and even up to 50% editing, showing that our EONs actually can achieve high double-digit editing in vivo. So in summary, we are again showing very consistent editing in the liver in these 3 different models with approximately 50% editing, including in vivo results. So that is a very nice basis to take our developments and learnings into the next step into developer. As the liver will be our target organ for us in our product development, we will use a proven modality called GalNAc to deliver our EONs to the hepatocytes in the liver. GalNAc is really a conjugate which is touched to the EONs and allow us to increase cell uptake, specifically hepatocytes by specifically targeting a receptor, which is called Asialoglycoprotein receptor, which is found on the surface of hepatocytes. And I'm going to show you a little bit more about the effect of such conjugation in our models. On the left, you can see the biochemical editing assay in which we test the capability of the EONs to add it at a certain position incubating the EON with the target RNA in combination with the ADAR molecule, then we can test the A2I performance of such molecules. You can see also that the addition of GalNAc to such EON does not interfere with the reaction at all, as you can see on the left. So we concluded that ADAR does allow for GalNAc to be present on the molecules. We took those lessons into a vivo study, as you can see on the right, and we tested a GalNAc EON combination and compared to a EON without a GalNac, as you can see over here. We dosed these animals 4 times with 10 milligrams per kilogram and then after 14 days, we tested the editing potential. And as you can see, we observed a tenfold increase after using the GalNAc conjugate as you can see over here. This reiterates and shows that GalNac can be actually used to enhance the uptake and efficacy of the EONs that we are using right now. And we will use that going forward in our development. So on this slide, I would really like to summarize what you've just seen in the slides that I presented. Overall, we can report consistent RNA editing in all the models that we've tested. And as you can see on the left, we have upto 40% editing in the CNS of mouse in vivo. We have achieved 50% editing in the liver as well in vivo. And of course, last but certainly not least, we have about 50% editing reported in the nervous system of non-new primates, which bodes well for the next steps in our development. In addition to that, we've tested and shown that the use of GalNAc can really help us to efficiently target RENs to the hepatocytes, which are the appropriate cells for our further development into liver targets. So now that we've looked at the Axiomer proof-of-concept data in general and the promising results that we've obtained there, we formed the basis look further now to focus on the broad therapeutic potential of our platform. On this slide, you can actually see how we envision the use of Axiomer to develop a novel class of medicines in different therapeutic areas. On the left side, you can actually see that we can actually correct mutations. There are many, many G2A mutations in the literature that we can actually use the Axiomer to develop a therapeutic to correct those. But maybe even more excitingly, on the right, you can see protein modulation because that's a very broad field that we can change the properties of the specific proteins with the purpose of developing therapeutics. For instance, we can alter protein function or to include protective variance to actually show and achieve loss or gain of function proteins helping to address or prevent disease. On the middle, you can see that we can actually interfere with post-translational modifications like glycosylation, phosphorylation, what have you. And those are important to regulate protein activity, preventing immune escape and slowing down degradation as well. And on the right, we can choose to use the technology to change protein interaction and that actually has a bearing to changing localization, folding, protein function and what have you. Specifically, I'm going to give you some examples of all of these applications on the next slide. So showing this would actually allow us to think about using the technology into broad therapeutic areas like common diseases, also rare diseases but also to target a wide variety of target organs, so to speak. Here, I would like to show you 2 examples of G2A mutation correction which were done in our labs. On the left you can see actually we picked a gene, which is mutated in the brain. As I can remember, I can remind you of previous result that we were able to after a single injection, we're able to get a 40% editing level in the brain. And on this particular site, you can see that after that, so 4 weeks after the single dose, we were able to correct the protein function up to 26 fold, and that is something that is very impressive. On the right panel, you see another example of a G2A correction, and that is in the CEP290 gene. And in organoids that we cultured, we were able to after 2 weeks of culture, after a single treatment, we were able to get a very significant correction of the protein, localization and function, again as you can see over here. So we have 2 examples of G2A mutation corrections that we can actually show protein correction as you can see. So the second example that I'd like to give you is that we are able to include a protective variant into the target messenger RNA that alters the function of a protein, and that's something that is shown over here, where we use PCSK9 as our model. So pro PCSK9 use autocleavage at position 152 to mature into PCSK9. So in patients, PCSK9 increases LDL cholesterol, so inhibition of autocleavage would lower PCSK9, resulting in a lower cholesterol load. So we turned to genetics and found a family with a very low level of PCSK9 and a very low level of cholesterol very, very healthy otherwise. So they have this very specific mutation at the autocleavage site, which actually results in a lower PCSK9 load. So thinking about that, we designed some of these EONs that could take care of a specific amino acid change in this autocleavage side and the result is shown on this particular slide. On the left, you can actually see that after the editing reaction in these cells, we cannot go up to 25% editing in this particular experiment using our DBCR. That in turn resulted on the right panel in about 80% reduction of PCSK9 secretion and also led to a shift in ratio from [indiscernible] PCSK9, as you can see over here. So it changed from 70-30 to up to 25% to 75% in treated samples. So the inability to undergo autocleavage likely retained the proenzyme in the endoplasmic reticulum where it acts as a dominant negative protein, preventing the exit of mature PCSK9. So this actually shows that with a change in an amino acid, you can interfere with the progression and maturation of a specific protein. So that is one of the second bucket. The third bucket is really to show that we can interfere with post-translational modifications. In this particular example that I'll show you here is that both translational modifications are really very much important for function of the protein such as stability, localization and certain EON channels depend on it whether they can open or close, depending on the editing of phosphorylation state. So protein phosphorylation or dephosphorylation is a PTM consisting of the addition or removal of phosphate groups to specific amino acids, which reside in proteins. So we selected an undisclosed protein, which is phosphorylated for its function, and we designed some of the EONs to actually alter the phosphorylation side. So after the editing, we could show, and this example shows you the ratio between phosphorylation and non-translation that we had a very significant reduction of over 25% of phosphorylation, which really opens up the way to manipulate the function of these proteins after treatment. So we will study that procedure much more, and we are going to be updating you as we go forward. But exactly, this is the second example for protein modulation, as we are being able to -- as we can perform on these particular proteins. So the last example that I'm going to give you is really to change a protein interaction with another ligand. So in this particular example, I'm going to tell you about protein interaction and protein-protein interaction or protein other ligand interactions are very common and important for function of certain processes. We select an example of such interaction and ANGPTL3 is an inhibitor of lipoprotein lipase and requires heparin binding for its function. We found in genetics, in human genetics, specific variants that are that have certain SNPs or variants in the heparin binding domain of ANGPTL3. ANGPTL3 with such variance actually leads to activation of LPL because of the fact that heparin-binding is impaired in those areas. So we designed EONs to mimic detect this change in heparin binding domain and tested that. On this slide, you can see the results. So after editing the ANGPTL3 with the change in the heparin binding domain, we could actually show an 80% decrease in heparin binding. Moreover, it seems that lower level editing in this particular case did not result in dominant decrease in heparin binding, but a higher degree of editing could actually show an 80% decrease in heparan buying. So very promising for the next step. But with this example, I can show -- I could show that by changing a specific, very important interaction, we could change the functionality of a protein as well. In summary, we think that the platform has brought applicability. We show that the potential is there to correct G2A mutations, and there are many monogenic diseases that are caused by this particular mutation, allowing just to use a G2A correction to restore the wild-type protein again. We have discovered the potential to modulate protein function to include a protective variant or to change the post-translational modification, but also to change, being able to change protein interactions with other ligands to create a phenotype in these proteins. So we feel that all of these examples lead to a broad therapeutic potential of the platform and will allow us to develop the products for rare and more common diseases. Okay. With this presentation, I hope that we've shown the consistent editing of all the platforms that we tested in the nervous system and in the liver, including non-human primate data. We were able to show that the addition of GalNAc allows us to efficiently target the EON to the hepatocyte and increase the editing efficiency. And by showing the different examples of the application we feel that we have a very broad applicability of our technology and that will provide us with a means to develop new products into rare and more common diseases as well. Thank you very much for your attention. And with that, I'm going to hand over to Rene for the IPO review and partnering strategy.

Thank you, Gerard, for the deep dive on the progress that we're making with our Axiomer platform. I will give you a short break and a breather. Steve Jobs was famous for many reasons. One being practice of one more thing. He would often wait until the end of the presentation to share one more thing, which was often a big announcement. Today, the one more thing I'd like to share with you, I will not say for the end of my presentation, but say it now. If you want to use an oligo to recruit ADAR, ProQR is the place to be. We have design and importantly, as I will highlight, we have the IP and a partnership strategy where we are open for business as points of proof for our leadership in RNA editing. We have been building an IP portfolio since we began working on Axiomer in 2014. And therefore, we are confident that we have a leading IP position to support our proprietary ADAR mediating RNA-editing platform technology. Back in 2014, I shared an office with Bart Klein, who is today our Senior Vice President of Axiomer Strategy. Bart is on top of a seasoned patent attorney also a brilliant scientist and wrote our first patent application in 2014 in my presence. And this was the basis of our IP estate, which protects all the foundational elements of the platform using endogenous ADAR for therapeutic purposes beyond 2040. Initially, Bart here came up with the idea to use natural known ADAR attractors to attach to oligonucleotides in the form of stem loop structures. This appeared to be working just fine, including recruit endogenous ADAR . However, these oligos were still rather large, and then in 2016, Bart worked together with a few other procurement inventors to achieve the same recruitment of endogenous ADAR but without the need of a stem loop structure. This allowed an even better opportunity to use the oligos for clinical purposes, simply because they were much smaller. And these are the oligos that we are developing today and have a solid IP position on to back up this early work. Patents have been granted in the major jurisdictions such as the United States of America and Europe for both types of technologies as indicated in the last column. The others are in various stages of examination and are expected to be granted in due course. In summary, we have 10 published patent families currently comprising of 22 patents, and we are working hard, very hard. We continue to invest in and expand our IP estate to maintain our leading position. We recently announced the successful defense of a key Axiomer patent protecting ADAR-mediated RNA editing. Your positions were filed in February 21 with the European patent office by 2 separate strawman against our granted patents, which is related to targeted RNA editing using endogenous ADARs. The opposition division of the EPO held a public hearing on March 7 and 8 and ruled in favor of our position after minor amendments of the main claim and 1 dependent claim. These nonmaterial modifications didn't weaken the claim patent at all. These claims were amended such that the oligos were limited to being chemically modified. In general terms, not specific to chemistries, to be completely novel over the prior [indiscernible] . This successful defense confirms that our IP is likely seen as problematic to others. Why otherwise would multiple parties try to oppose a patent position, spending valuable time and money. Given the importance and value of our intellectual property estate, we intend to continue to defend against these challenges and remain confident in a leading position. So one more thing, ProQRs Axiomer leading IP portfolio is robust, broad and provides coverage for the fundamental features of the technologies beyond 2040. We now turn to our partnership strategy. We believe our IP along with our deep expertise in RNA editing and oligo therapeutic approaches is a source of great value to potential partners. For ProQR, a key part of our strategy is the ability to selectively form partnerships. They bring important resources, capabilities and funding to further enhance our programs. But they also advance applications of the platform beyond the targets we intend to develop independently, creating a new class of medicine based on Axiomer. We closed our first Axiomer partnership with Eli Lilly in September '21, expanding at last December, bringing the total potential value to approximately $3.9 billion plus royalties. The expanded collaboration is important validation of our leadership in ADAR-mediated RNA editing our boost IP and the potential of a broad applicability of the Axiomer platform. Lilly has the option to expand the partnership further with another 5 targets for an additional $50 million opt-in fee to reach a total 15 targets. I would say this is a very, very pleasant sword of Damocles dangling above our hats, and we look forward to continuing to progress the productive partnership with Eli Lilly improving the lives of patients together. Now a few words on our ophthalmology programs, which do not utilize Axiomer. We are encouraged by the progress that we're making together with Lazard to identify a partner who can continue to advance these programs, and I have great expectations finding new homes for these assets. And of course, when there is an update, we will certainly share this with you. Having said all that, beyond our partnership with Eli Lilly, we do have appetite and capacity to further solidly partner our Axiomer RNA editing platform technology. With regard to that, we are seeking research collaboration partnership that could take the form of single or multiple asset licensing agreements or broader therapeutic or organ-specific transactions. We are right now focused on dialogues around liver, extra hepatic and metabolic disorders as well as rare neurological diseases with an unmet medical need. The potential is broad, extremely broad, as highlighted on this slide. We continue to be motivated to execute value-creating transactions, harnessing ProQR's leadership position in precise RNA editing and endogenous ADAR recruitment, given the fast potential of Axiomer to target a broad range of diseases, we look forward to keeping you abreast and updated on developments with our partnership strategy. Now before I turn the presentation back to Gerard, who will present our pipeline overview, I would like to highlight that the shop was opened for business during renovation last year. With Sarah, Daniel and myself since I rejoined in June last year, almost weekly investor calls and visits. And I promise we intend to continue to do so. And I realize that there is a lot to take in from today's presentations, and we are happy -- very happy and honored to take continuous one-on-one meetings with a virtual or face-to-face, to further elaborate and interact with you and present ProQR as an investment case. As a matter of fact, we are in New York in the first week of April. So if you're interested, please reach out to Sarah. And with that, without further ado, I would like to give the floor back to Gerard.

Yes. Thank you, Rene. I think we will now focus on the in-house pipeline development for ProQR and then the next slide, I will take you through the first targets that we will be developing going forward. As can be -- as seen in the previous sections, ADAR editing is a very active mechanism, which is widely present in the whole body. As such Axiomer platform really takes advantage of the natural activity and has a broad applicability basically with the possibility to target multiple organ systems, as you can see over here. For our initial pipeline products, we are going to be focusing on the liver, and we feel that the liver is very attractive as a target organ for Axiomer for really multiple reasons, which are shown on this slide. A very obvious one is the high editing potential in the liver, and I hope I have convinced you of that in my platform slides previously. We can actually enhance the liver targeting by using the GalNAc technology, which has been proven in previous developments for using oligonucleotides as well. 85% in the liver consists of hepatocyte and they have the asialoglycoprotein receptors, which is targeted by using GalNAC. In addition to that, we've shown that GalNAC does not interfere with the function of ADAR itself. So in drug development itself, biomarkers are essential to understand target engagement and also to assess efficacy and safety as well. The presence of biomarkers as such allows us to monitor and see the target engagement that we will have with our Axiomer technology, and that's what we'll be using moving forward into development as well. The liver is at the center of a high metabolic activities and it does influence other target organs. The high metabolic activities as you can see, involve bile acid production and excretion, lipid production and plasma protein synthesis to name a few. There are many diseases that originate from the liver and by delivering medicine to liver, we can target multiple organs as you can see over here. The numerous liver originated diseases are listed over here. And to name a few are cholestatic diseases, metabolic disorders or cardiovascular diseases or CVDs. As Daniel mentioned before, our rationale is to go after new targets guided by human genetics. And I'm going to show you details of that in the 2 therapeutic areas that we've decided to pursue. I'm going to walk you through the rationale of the initial targets and give you some more information on the disabling diseases that we plan to investigate. Let me turn to AX-0810, which is our first pipeline program and it's designed to target cholestatic diseases. So in short, when there is an excessive accumulation of bile acids in the liver that can lead to cell stress and damage, and we call those diseases cholestatic diseases. On the left, you can see the liver cells that are actually at the center of bile acid production and bile acids are derived from cholesterol metabolism. Once the bile acids are produced, they are stored in the gall blader but excreted from the liver to the intestines. There, the bile acids facilitate the digestion of fat, nutrients for growth, fat-soluble vitamins and carry away waste. Most of the bile acid, some 95% return into liver via reuptake from the portal vein. On the right, you can see that individuals with cholestatic diseases actually have a dysfunctional bile duct that causes bile acid accumulation in the liver and the buildup of bile acid is really toxic and leads to inflammation and liver damage. On this particular slide, you can see that without treatment, the accumulation of bile acid in cholestatic disease leads to progressive liver degeneration, starting with liver inflammation, progressing to fibrosis and then, unfortunately, leading to liver failure, malignancies and poor life prognosis. Out of the 2 -- out of all the noncholestatic diseases, 2 of them are leading conditions for liver transplantation in adult and pediatric population, primary sclerosing cholangitis or PSC and biliary atresia or BA. So here, you can see the high unmet medical need, which still remains in PSC and BA. PSC is a condition that is being diagnosed at the age of 30 to 40 years with a higher prevalence in men. There are about 80,000 patients in the North America, Europe and Japan, and it's autoimmune liver disorder leading to bile duct stricture. BA on the other hand is a pediatric condition affecting about 24,000 individuals and it's very rapidly diagnosed in the first weeks of life and caused by congenital absence or defective bile duct. Both of the conditions are associated with symptoms that have a huge impact on quality of life, such as pruritus, fatigue, poor weight gain or weight loss. And of course, they have a rapid progression into cirrhosis and liver failure. So both PSC have and BA have a high unmet medical need since there are no approved therapies available at this time. For PSC, the only approach currently available is liver transplant. And as such, PSC is known as the leading autoimmune indication for liver transplant in the adult population. And unfortunately, 20% to 40% of the patients present with PSC recurrence after liver transplant. For BA, newborn babies can undergo a very complex surgery, which is called hepato-porto-enterostomy or HPE, for which a surgeon removes extrahepatic duct bile -- bile duct, excuse me, for the infant and links the liver directly to the intestine to remove the bile acids. Post operation procedure, a majority of children will eventually develop portal hypertension and require a liver transplant before reaching adulthood. On this particular slide, I'd like to present with you the solution that we have in mind. And as I've mentioned previously, we learned from [indiscernible] genetics to inform our pipeline development. I turn to sodium-taurocholate co-transporting polypeptide also known as NTCP, which is actually the main transporter involved in bio acid reuptake into the liver. As cholestatic diseases are caused by accumulation of bile acids in the liver, altering the reuptake could have a positive impact on the course of such condition. And interestingly, it has been reported that loss of function variants of NTCP naturally occur in some people with only a mild phenotype without any comorbidity. Furthermore, pharmacological modulation of NTCP has been shown to improve outcomes in a mouse model of cholestasis, reducing inflammation and liver damage. So we have consolidated our learnings from scientific data in human genetics into a novel therapeutic AX-0810 targeting NTCP is designed to reduce bile acid reuptake into the liver. It's an editing oligonucleotide that will attach or binds to the NTCP messenger RNA and will create after editing a loss of function variant that will no longer be able to perform the task of reuptake the bile acids into the hepatocytes. On the right, you can see that as such, AX-0810 aims to reduce the bile acid reuptake and reduce the bile acid load into the liver. With this program, our objective is to alleviate symptoms in PSC and BA and to prevent or delay the progression of liver damage and need for liver transplant. To ensure success of this program, we have a rigorous and pragmatic approach that we think will increase our chances of success to commercialization. On the left, you can see that we have a robust preclinical model package available with a possibility of assessing proof of mechanism early in models. We have the complex organoid model that actually has all the machinery available to study bile acid synthesis and uptake and reuptake in our in-culture as well as animal models that have bile duct stricture conditions as well. The program does allow to an early start clinical trial in healthy volunteers. As such, we can study that timely insight and safety and target engagement with validated biomarkers that we'll use. So for instance, we will be looking at bile acids and serum, liver enzymes like ALP amongst others. And of course, we have a plan moving forward into later-stage clinical trial with disease specific endpoints and a clear regulatory pathway. As mentioned on the previous slide, there is the possibility of generating early clinical insights via Phase I in healthy volunteers with the objective to assess safety, tolerability and pharmacokinetics and dynamics of our molecule without actually having been bothered by the account commitment pathological conditions. We also plan to measure RNA editing and circulating exosomes in plasma that will allow us to get an early insight in the target engagement of our EONs. The aim is to develop one clinical trial to ensure timely recruitment and data generation and the trial design will include single and multiple dose ascending cohorts and target an entry into the clinic by late 2024 or early '25. So in summary, AX-0810 is a first pipeline program as being developed a high unmet medical need in cholestatic diseases with PSC and BA being the lead indications. It's a novel and untargeted approach from originating from human genetics and with the objective to reduce bile acid reuptake in to liver through NTCP loss-of-function variants. We have a rigorous approach to increase the probability of success from preclinical to clinical stage, including use of biomarkers, which are validated and a clear regulatory pathway. And the next step, will be the generation and selection of our lead candidates for which the data will be presented in due course at scientific conferences. We anticipate entry of clinical trials late 2024 or early '25. Like I said, Axiomer can address numerous liver originated diseases, and we'll be looking now into our second pipeline program, as you can see over here. This example is a clear example, Axiomer has brought the development potential by targeting the B4GALT1 protein, which is highly present in the liver and can have a positive impact on risks related to cardiovascular diseases. Okay, we have selected AX-1412 as our second pipeline product to address the remaining needs, to reduce residual risk factors for cardiovascular diseases. We aim to develop this program up to the early clinical stage. And after the first clinical data has been obtained, we will partner with another company for further development. Cardiovascular diseases or CVDs are really a group of health conditions that affect the heart and blood vessels such as atherosclerosis, which can lead to severe problems like heart attacks, heart failure and stroke and are the leading cost of disability and death globally becoming a significant health issue. Shockingly, there are around 80 million people dying from CVDs each year, making up 30% of all deaths globally according to a report by the WHO in '21. In the United States alone, the American Heart Association estimates that by 2035, more than 130 million adults will have some for form of CVD. And despite the standards of care, the medical need remains, even with existing therapies, less than 35% of Americans with high LDL cholesterol levels reached their target levels recommended by our guidelines. And CVD events still occur even when LDL cholesterol levels meet clinical goals. Many patients also struggle to continue taking their meds 2 years after a CVD event. And in addition to that, about 5% to 10% patients cannot tolerate high doses of for instance, statins primarily due to muscle aches. LDL, there are certain independent risk factors involved in CVDs that may have a negative synergistic effect. So LDL cholesterol is reported as a major risk factor for CVD, increasing the risk of arterial plaque formation as you can see on the left of this cartoon. Another risk factor reported in literature is fibrinogen and its increased risk for blood clotting. Furthermore, LDL-cholesterol and fibrinogen are 2 independent risk factors involved in that may have negative synergistic effects on the risk of cardiovascular diseases. LDL cholesterol risk factors is being addressed by current standard of care but none of the treatments on the market specifically are focused to address the both as you can see over here. And interestingly, it has been reported in literature as well as human genetics that have an old order Amish-enriched missense variant in a functional B4GALT1 beta galactosyltransferase protein was associated with lower LDL cholesterol and a lower plasma fibrinogen. So B4GALT1 is an enzyme that transfers [indiscernible] from uridine diphosphate galactose to specific glycoprotein substrates like apolipoprotein B and fibrinogen. Furthermore, a specific loss of function variant in B4GALT1 has been reported to be associated with decreased coronary artery disease in a gene-based analysis. So it's learning from that protective variant, which was reported in human genetics that decided us, that made us decide to develop AX-1412 as an EON, which leads to a loss of function in of B4GALT1 as described in literature. The B4GALT1 loss of function variant induces hypogalactosylated of the lipoprotein B-100 [indiscernible] and limit their negative effects. It is a novel and unique approach, which can address simultaneously 2 cardiovascular risk factors, and that's really not suitable for knockdown technologies as shown in literature leads to [indiscernible] and a severe developmental anomalies in studies. AX-1412 can lower LDL cholesterol and fibrinogen levels to reduce residual risk in cardiovascular diseases and prevent or delay the development of cardiovascular events. As presented with our first pipeline project, our development approach for the second program also meets all the thresholds we've put to increase probability of success, including a very strong preclinical model package available with good translatability into the clinic. A timely early-stage clinical Phase I that will give us some initial insights into the clinical effect and we're going to go after conditions with disease-specific endpoints and a good guidance from regulatory agencies . As mentioned in the previous slide, there is the possibility, like for our first pipeline project to generate early clinical insights via a Phase I style in healthy volunteers. This is with the object to assess safety, tolerability, pharmacodynamics and kinetics of our program AX-1412. This without interference of co-commitment pathological conditions and to investigate target engagement and [indiscernible] effects through specific biomarkers. We will also plan to measure RNA editing through exosome analysis, that will give us a very early insight into target engagement at the RNA level as well. We aim to have one clinical trial to ensure timely recruitment and data generation and the trial design will also include single and multiple dose ascending cohorts and target an entry into the clinic in late 2024 or early '25. So in this summary slide and the next steps for our program, I can reiterate that although there are several approaches to lower the risk of cardiovascular disease including reducing LDL cholesterol and [indiscernible] levels, reducing fibrinogen levels may offer additional benefits to patients with unmet medical needs in this large population. AX-1412 is novel and unique approach originated from human genetics that could have pleiotropic effect for cardiovascular protection and is really not suitable for knockdown technologies. As our second pipeline program, we have a rigorous approach to increase probability of success from preclinical to the clinical stage, including the use of validated biomarkers and next steps will also include the generation and selection of a lead molecule for which data will be presented in due course at scientific conferences. We aim to enter the clinic by late 2024 or early '25. And with this program. So in summary, I'm very excited to have announced our 2 development programs, and we will pursue to develop liver originated disorders for these programs as we move forward. But we feel that the broad applicability of our Axiomer platform does not limit us to target one organ only. And we aim over the next years to broaden our pipeline, targeting metabolic conditions, rare neuro disorders and potentially other conditions that we will not disclose today. We do have a very dynamic thorough and robust capacity to find new targets suitable for Axiomer and we will continue to push the boundaries to develop new therapies using our technology as we are just scratching the surface of our Axiomer potential and our beginning of a new era, the Axiomer era. I will now hand it over to Daniel for the summary and milestone section. Thank you very much for your attention.

Thank you, Gerard. We're confident that we have put ProQR in a strong position to execute on the business plan to capture the full value of the Axiomer opportunity. We believe this approach holds great promise, and we see RNA editing as the next evolution in RNA therapies. And Axiomer can contribute by creating a new class of medicines with broad therapeutic potential. Today, we announced AX-0810 for cholestatic diseases targeting the NTCP channel and AX-1412 for cardiovascular diseases, starting the B4GALT1 gene as our initial pipeline programs. These programs along with the other targets in our pipeline, share several key features, a deep routing in human genetics, the potential to have a major impact in a high unmet medical need. The ability to leverage the existing proof and delivery technology, the opportunity to establish target engagement via biomarkers in early clinical testing as well as well-defined clinical end points. We plan to advance these programs into the clinic in late 2024 or early 2025. The platform has shown consistent RNA editing models in the nervous system and liver, including in vivo data demonstrating up to 40% editing in a nervous system of mice leading to a 26 fold change in protein function. We've also seen up to 50% editing in the liver of mice and up to 50% editing in a nervous system of nonhuman primates. These effects were durable with editing and protein activity reported 4 weeks following a single injection. We also showed that GalNac does not interfere with A-to-I editing and that it increased the editing efficiency by tenfold in [ in-vivo ]. Axiomer has shown broad applicability and proof of concept by mutation correction with protein recovery by generation of variance in wild-type sequences, also by changing protein function with dominant negative effect by regulating translational modifications and by impacting protein-protein interactions. This opens the door to many more applications beyond what we have discussed today. We believe that Axiomer can potentially lead to hundreds of new medicines. As we have seen today, a lot of exciting progress lays ahead in the platform and pipeline. After this update today, we will continue to share more platform data as the time progresses. This will include additional data presentations at scientific conferences and publications in peer review journals. Furthermore, we plan to share additional platform data over the next 12 months, including further nonhuman primate data with delivery to a better sites in liver. And as we progress, we will also provide further insights in the pipeline and discovery progress to populate rich pipeline that captures the broad Axiomer opportunity. Then on the pipeline. We will, over the course of the next 18 months, share several updates with translational data on our initial 2 clinical programs. This will include nonclinical proof-of-concept data updates with supporting, which will be supporting these programs as well as several translational data updates that will help you to understand the translational derisking strategy into the clinic. Subsequently, we plan to advance our 2 programs, AX810 for cholestatic diseases starting the NTCP channel and AX1412 for cardiovascular disease, targeting the B4GELT-1gene in late 2024 or early 2025. We will continue to execute with high priority on our Lilly partnership and work towards to opt in from Lilly that we anticipate this calendar year which would come with a $50 million payment to ProQR. And we expect to enter into another 1 or 2 multidiscovery partnerships as we progress. We will continue to further enforce our appear states as we plan to preserve and expand our leading IP position. And with the cash on hand, we are well funded to execute on all that we have discussed today with a runway into mid-2026. Today, we have highlighted the progress that we have made to advance our proprietary Axiomer RNA editing platform technology. We also demonstrated its broad applicability and how we plan to develop treatments for patients that suffer from diseases with high unmet medical needs. We have the science, a platform approach with broad applicability and preclinical proof of concept across multiple models and targets. We're now advancing our initial pipeline programs. Our leading IP estate and our recently expanded partnership with Lilly are important validating points, and we have an experienced team executing on this strategy. And as a company, we are well funded with a cash position that funds us into mid-2026, to execute on the Axiomer business plan we have laid out today. ProQR is leading the advancement of the RNA editing field as a new class of therapies for patients. I want to thank you all for joining today to learn about our Axiomer platform and pipeline. And I will now hand the call back to the operator for questions.

[Operator Instructions] With that, the first question will come from Steve Seedhouse at Raymond James.

And few that I'll work through some science ones and then some strategy ones. Just first on the science. I'm curious about the turnover of an exogenous editing oligonucleotide. And how you would compare or contrast that with the turnover of RNAi or RNA H for siRNAs or ASOs? And are there any implications, if there are differences or the dose that you would need to use in humans, is it basically going to be in the same ballpark as an oligo that would be used for knockdown or splice correction.

Yes. Steve, thank you for the question. Thanks for attending the event. We're going to have the science questions be addressed by Gerard.

Thanks, Steve, for the very good question. I would say that the oligonucleotides that we are developing for our editing reaction to EONs are quite similar to the ones that we developed with a full modification for the earlier, let's say, spice [ modulation ], somewhat different, but still similar. So I would say that the actual dosing frequency. We have now data that goes out to, let's say, 4 weeks and beyond, with the initial experiments that are ongoing, are focusing on to see how long the effect last, but we anticipate, let's say, a quarterly or maybe a twice-yearly dosing that will be envisaged for these EONs. So I hope that answers your question.

Yes. And maybe just on cholestatic program, a really interesting approach. And there's a few of these diseases that via IBAT inhibitors reducing bile acids has proven to be effective, some pediatric indications and arguably PBC as well. You've kind of highlighted biliary atresia, PSC in fact, as maybe the first to indications that you would pursue for that mechanism. I'm curious if that's more of a sort of commercial unmet need decision and you're sort of anticipating proof-of-concept coming pharmacologically in those indications in the coming year or so before you're in the clinic? Or do you think the mechanism is actually better suited for those diseases on a scientific or medical basis.

Right. Again, a good question. I think the focus of our program that will tackle the PSC or BAs life has a basis that we are going to be targeting the disease on target in the hepatocyte where the primary problem lies. So the -- let's say, the faulty bile ducts expiration of bile acids and the high influx of bile acids through NTCP is actually causing the problem and then the progression from the liver into, let's say, the late-stage progression into toward the liver transplantation. So we feel -- so the IBAT inhibitors target the enterocytes transporters, whereas we are going to be focusing to limit the reuptake of bile acids in the hepatocytes. So that is a more direct way of dealing with the problem. And looking at human genetics, we found that there are people out there with a loss of function variant of the NTCP that actually gives rise to a lower uptake into the hepatocyte. So learning what the nature has given us, we are going to apply that to the problem, which we find in the cholestatic disease.

Could I ask just on this program? If -- like should Lilly, for example, opt-in on an additional 5 targets, is this -- could this be among those targets that are partnered with Lilly, or is this carved out now and going forward as ProQR is wholly owned.

Yes, Steve, I'll take that question. So this is [indiscernible] will be ProQRs program going forward. So we are in control about the future of these programs that we have announced.

Great. Last question for me. Just -- I was hoping you could comment on just the path that the clinic over the next 2 years. My guess is the market today is sort of expressing some [ ambiguous ] to the timeline of late '24, early '25. And so it might be helpful if you could just provide some context for sort of why that is and what boxes you need to check and sort of maybe when these programs became defined by you as lead programs and maybe that will help just provide some context and comfort on the timeline.

Yes, absolutely happy to. So obviously, we are working on a really exciting novel technology that allows us potentially to make dozens, if not hundreds or thousands of different medicines. We're very mindful of the fact that the first few programs carry the burden of successful translation of the wider platform and unlocking that value into the clinic. So we're really mindful of not cutting unnecessary corners by really building a solar translational data package that derisks our steps into the clinic which allows us to go into the clinic with high probability of success as the importance of the validation of this platform in the clinic with these first molecules is so significant. So we are in the next 18 to 24 months going through a number of different steps to move these programs towards that first clinical step. That does include additional proof-of-concept work that we will announce over the next 12 months that will allow us at scientific meetings to share this and provide further context around these programs. Over the next 18 months, we will provide additional translational data that will give further insights in the derisking translational strategy, so how these molecules are preclinically derisked to translate into the clinic. And subsequently, we move into the clinic. I think as Gerard laid out in the slides today, 1 of the key criteria in our selection of these targets was that we can test these targets in healthy volunteers, which will make the clinical execution much more straightforward in terms of timelines, in terms of execution, in terms of cost, and will give us a data set that will early on in the clinical development, give us high confidence in terms of target engagement of these molecules in human. So all designed really to maximize the probability of technical success of translation into the clinic. And there's quite a bit of news flow you can expect from that.

The next question comes from Jennifer Kim at Cantor.

I have a few questions. Maybe to start off, to follow up on -- with the last analyst ask -- could you lay out how soon we could get early clinical data in humans? And then also on the path to translational and proof-of-concept work. I know you said over the next 12 months, but is there more latter half? Or could that trickle more into starting in early 2024? Maybe we'll start there.

Yes. Jen, thank you for your questions and for attending the event. So we're guiding to the next 12 months. But in the next 12 months, we're going to have multiple data disclosures through scientific presentations at scientific conferences. We're going to do peer reviewed journal publications. We're going to be talking a lot about the programs and providing initial insights, both proof-of-concept data as well as translational data. So we're not guiding to this year or next year, but it's going to come over the next 12 months, spread out across multiple different disclosures. And sorry, your other question was?

A follow-up question. I think you talked about being interested in 1 to 2 sort of multi-target discovery partnerships as you progress. Do you believe that's a 2023 focus? Or from your conversations that you've had with potential partners, do you believe that there's anything specific that those potential partners are looking for before pulling the trigger.

Yes, that's a great question. So we are looking at potentially doing 1 or 2 additional business development, multi-target discovery deals over the course of this year and next year. We think that we have the band way to execute on that. Certainly, the platform is broad enough to not cannibalize on the opportunity on the platform by doing 1 or 2 additional these transactions. We actually through the partnership with Lilly against some really important insights. We've seen how important the partnership has been for us as a company. And yes, the Lilly team has been great to work with. So we plan to expand that and to go further on to the strategic side as well. That could happen over the course of this year. We obviously -- with these things can have firmly guide to these events, but it's certainly our aspiration.

Okay. And then maybe one last question. You've been pretty consistent Highland Game. Can you remind us of whether -- or how you're leveraging your partnership with Lilly on that front? And have you made any decisions there?

Yes. Great question, Jen. So as part of the partnership that we have with Lilly, we are also granted access to their delivery technology. So we can, on a case-by-case basis, use specific delivery technology that Lilly developed and validated for programs that we develop in our wholly owned pipeline. That, for example, does include the GalNAc that Lilly uses. So we have the possibility to use that selectively for the targets that we bring into the pipeline as we progress forward. We do believe that GalNAc is the way to go. We think GalNAc has been tested and proven, and it's certainly the most straightforward way to get preferential delivery to the peptides. So likely, we will use GalNAc going forward for the programs translating those into the clinic. And that maybe the Lilly GalNAc we'll come back to that at a later time.

The next question comes from Alexandra Van Harton at Kempen.

I asked some questions actually. Firstly, you already elaborated a bit on the news flow for the coming year. But I was curious, are you also planning on the announcement of additional programs? Do you desire to have a certain cadence of new programs?

Yes. Thanks, Alexandra, for the question. So yes, we do have a rich news flow for the next 12 months and beyond, both on the strategic [indiscernible] side as well as on data, translational data and the pipeline. We may decide over time to advance additional programs going forward, but we're not guiding to that right now. There's 2 progress that we now plan to use validate the platform in the clinic -- and those will be prioritized and those will be the first data points to show. In the meantime, we will obviously progress broader on the pipeline as it will help us to further build on the internal pipeline as well as on the potential business development front. So there is additional progress to be made there, how they'll fit into the new flow remains to be seen. I think you had a second question, right?

Yes. It's very clear. And then I have another question that's more on the scientific side about a very impressive editing efficiency that you showed for actin B across in vitro and in vivo models. I was curious, how does this efficiency compare across different genes and specifically your announced target genes? And is there a lot of variability there.

Thank, Alexandra, that's a great question. I think that we started earlier during the time that we started to understand what the ground rules were, and we now understand how we design these EONs for differential editing and the editing of the actin B, which is a housekeeping gene is very robust over the different systems, as you've seen neuronal cells, hepatocytes the complex organoid systems that allow us to really study the actual editing in the let's say, the surroundings that we want to see it in, but also in the in vivo situation. When we take these learnings into other genes that I've shown before, we do see that there's a portability in the concept. So we take the learnings that we have from the housekeeping genes, developed our EONs, for instance, for the alpha-1 or the ANGPTL3, and we see robust editing as we move forward. So every target has its own challenges and optimization is needed. But all in all, the ground rules that we've taken in are portable from 1 gene to another. So that helps a lot. Hope that answers your question.

The next question comes from John Wolleben at JMP.

A couple for me. Just wondering about the data you presented preclinically from the [ road into ] the nonhuman primate. Do you think that these are at potentially relevant human doses? And then how should we think about the translation from 40% to 50% added into a potential benefit? What's -- is that enough editing efficiency? Or do you more -- just thinking about how we should translate this from the preclinical to the potential clinical data?

Yes. Thank you, John, for the question. Thank you for attending the event. I'll let Gerard to address your questions on the science.

Yes. So we think that the robust 50% editing really is a very good basis to address the, let's say, clinical relevance in the other models that we have. We feel that we are looking at human genetics to give us guidance on what we need to achieve. And we see in the models that we chose, the data shows that heterozygotes people which equals, let's say, 50% [indiscernible] already have a clinical benefit. But we do also have data showing that in our models, that's less than 50% we actually do the job and give us a good result that we would like to achieve. So anywhere between 25% and 50% would be my target to go after in a relevant program, that's the current thing.

That's helpful. And 1 bigger picture. How are you thinking about prioritizing internal programs? I guess the big difference between the liver program and the cardiovascular would be the size of the market and potential patients. But what other kind of attributes are you using to strategically pick what stays in and what goes out.

Yes. So John, let me comment on that. So we've obviously done a lot of work to triage all the opportunity that lays ahead of us with Axiomer. We see so many opportunities how Axiomer can be applied to generate new therapeutics that we developed a very comprehensive system to essentially prioritize these targets which led to the pipeline that we have announced today. I think important elements in that systematic approach to us are to prioritize targets that have deep routing in human genetics, so that we completely know that the, let's say, post editing RNA product will have beneficial effects on disease [indiscernible] in patients. Second, for this new technology, we have decided that we want to really study this mechanism of action in isolation of any other variables. So we have to have luxury position to be able to use an existing therapeutic modality. And therefore, we can use a lot of known [ molecules ] for delivery, for example, that allow us to -- with high probability of confidence -- with high confidence, deliver these molecules to the current organ and not have to worry about actual delivery of these molecules such that we can really look at the therapeutic effect. Then -- so for that, we picked the liver as a target organ as delivery to the liver is pretty much derisked. Then in a liver, we have, I think, the advantage that we can look at a lot of biomarkers in plasma. So that allows us to very early on and study target engagement of our molecules and the downstream effect of that in biomarkers. And that, I think, provides us a quick path to building confidence in the broader platform and individual programs. And then for these 2 programs, we actually prioritize targets that can be studied in healthy volunteers because we all know that studies in patients can be very lengthy. It can be complex to recruits, can be complex to have -- have a consistent pattern in the data. And in health and controls, there's obviously much less variability and much better to control in such a study in an efficient way. So those are some of the important matters. I think for what we will long-term keep for the in-house pipeline and what will be potentially used for individual product partnerships, largely has to do with the time and cost to get the product developed to market and the ultimate commercial infrastructure that's required to commercialize these products. So looking at the 2 assets that we today have announced and that will move forward into the clinic. And the product for cholestatic disease AX810 will be prioritized for our in-house pipeline to develop for AX1412, we will likely seek a partner after we have established clinical proof-of-concept in the first clinical trials, such that we can continue development there with the partner for the future. I think that pattern, you will see throughout the rest of our pipeline as we should progress.

Congrats on the progress.

The next question comes from Ashik Mubarack from Citi.

This is Ashik on for Yigal. I just have a few. Maybe first a more practical question. It sounds like you're expecting that you'll be able to generate some initial clinical data, at least within your guided cash runway. I'm curious if you can talk a little more about what specifically you mean by proof-of-concept, meaning data within healthy volunteers? Or do you think that timeframe is enough to show POC and natural patients?

Yes, Ashik, thank you for attending the event today, and thank you for this question. So we have [indiscernible] a pretty robust schedule runway as we have announced with our year-end cash today. We have a runway that is well into 2026 until mid-2026, and there's quite a bit of upside to the runway with potentially the $50 million opt-in from Lilly with potentially additional big deals on the Axiomer platform but potentially the best into the ophthalmology portfolio, all nondilutive ways to help us extend the runway significantly beyond that potential. So from a cash runway perspective, we feel pretty comfortable. Currently, we're guiding to start up these trials in late '24, early '25. So with that, you can count on having clinical data from these initial trials, certainly well within a run rate. If that includes patient data in a follow-up study that we are not guiding towards yet because it also depends on the learnings from the trials. What is most important, I think for us, Ashik, in validating this technology into the clinic is that we get reliable data that is answering the question on how efficiently we can edit in the human setting in vivo editing that is. And the trials that we're executing on are designed to answer those questions, while in parallel, advancing 2 potentially very important treatments forward on a development trajectory.

Got it. That's very clear. Maybe I'll ask one on ADAR itself. I guess as we think about this from a broader perspective, what's the rate limiting step within this editing process? Is it the recruitment and availability of ADAR? Or is it more to do with the dosing and persistence of your oligonucleotide, what's being the rate-limiting step? And maybe how efficient or consistent is the recruitment of ADAR, especially across your various experiments.

Those are great questions. I think the recruitment rate of ADAR and the oligo, we studied that in vitro and I think it's a very efficient process, to be honest. And I think that having learned a lot from our experiments, the rate limiting in the beginning was really to understand the chemical modifications that we could enter into these oligos that could actually be compatible with what ADAR could do. So basically, the chemical modifications that we introduced, not to enhance delivery but also stability we very carefully titrated those together with Dr. Beal to see what the balance would be between effectivity and delivery. So I now have learned a lot more including the use of GalNAc, I think the limitations start to disappear and the actual dosing and frequency starts to become more oligo like, if you will, that has been in the field of oligos. So we are understanding more about the chemical modifications, but the behavior of these molecules and what ADAR can actually tolerate in that. So I hope that answers your question.

Yes, that's very helpful. Last one for me. I think we have a good understanding of what you're looking for on the efficacy front, especially from a biomarker perspective, but what are you looking for on the safety side? I assume you'll maybe talk about potential liver AEs or something to that effect. But you also alluded to ADAR's role as an antiviral player. And I'm wondering if there are potential immune AEs you might be keeping an eye on -- an [ eye on ] or something to that effect?

So great questions again. We would look for toxicities that would be related to oligonucleotides. Our EONs basically are oligonucleotides that you would see in other mode of actions. So we would be looking at systemic events, so to speak, that would focus on liver and others that are known to the field. I don't think that we will see off-target effect on activating the immune system because that's a slightly different mode of action. So mainly, we will be looking at the oligo associated toxicities that are out there. But having, let's say, 4 decades of experience from the field in using oligonucleotides. We know very well in what to look for and what to select against. And that's something that it will be [ content ] center in our development programs moving forward.

The next question comes from Dae Gon Ha with Stifel.

Couple. So maybe I'll just go 1 by one. So Daniel, at a high level, just to follow up on an earlier I think it was Steve's question. I guess when you think about prioritizing programs, can you maybe tell us a little bit about how you're thinking the insights from these initial 2 programs, what kind of derisking they would provide for your subsequent programs, which remain undisclosed at this time? Is it just similarities in the organ targeting, the cell types that you're targeting, the nature of the edit? What would be sort of the main sort of take home from a read-through perspective.

Yes. Dae Gon, thank you for the question. So look, I think the first organ we want to unlock on the Axiomer platform is the liver. We think in the liver, there's a lot of opportunity to targets, proteins that are excluded from the liver and can cause or prevent disease throughout the entire human body. We think that with these first programs, we can really start to build a correlation model to understand how our models -- how our molecules translate through the nonclinical models into the clinic. And based on that correlation, we can extrapolate for future targets how they will behave, which largely has to do with delivery, durability, dosing -- editing efficiency, translation of editing efficiency into biomarkers. There are some disease specificity of course, around the biomarker aspect. But we think we will learn enough by changing a few variables from target to target to really be able to increase the probability of success for each following molecule on the basis of the derisking we're doing with the first few. I think we're trying to keep as many of the variables the same. So targeting a better sites with GalNAc conjugated molecules will allow us to really study the different targets in isolation of any other [indiscernible].

Great. Great. Maybe some science question since I'm a little bit naive on the ADAR front. So maybe Gerard, on the ADAR expression profile, if I could ask a 2-part question. Can you maybe walk us through the differentiated or differential expression profile in between cell types and between ages and between, I guess, organs or maybe in their disease conditions, do ADARs get expressed differently? And then on the other side, can you also maybe talk a little bit about ADAR subtypes whether they are constitutively expressed at a similar level or how you could go about designing EONs appropriately so that you are recruiting the type that you want for the output.

Right. Excellent question. I think that, in general, we spend a lot of time in understanding expression of ADAR as a class of editing molecules. And as you may know, there are 3 ADARs, of which 2 ADARs are interesting to us because they can help us to edit. You have the ADAR1, which is 2 isoforms, which is actually the 150 isoform and the 110 of which the -- 150 isoform is basically interferon inducible and it's basically cytoplasmic but also shuttles into the nucleus where the smaller form of ADAR1 is mainly nuclear and then you have ADAR2, which is nuclear as well. So we have an abundance of ADAR molecules that are in the cell. And we found that comparing different cell lines, there are some editing efficiencies that differ from cell to cell. But we find that. If you look at the hepatocyte, we have a very fairly well and consistent pattern of editing taking place. And that's that are learning from our systems that we took from testing the system in hepatocytes -- primary hepatocytes and then we took them into more complex systems, which we call LMT or liter micro tissues. We see a very nice portability of editing efficiencies that we can take from those learnings. And I think what we've seen is also taking that into Vivo now we can actually see that the -- well, there's a robust editing taking place in every system that we now test. So we do feel that there's a consistency around the editing taking place in the difference or cell systems that we're testing. So that helps a lot in the development and taking the next steps.

Got you. And then just 2 clarification questions. On one of the -- I think it was the spinal cord data in [ NHP ]. There was [ NF3 ], but with the 2 [indiscernible] saying 2 were excluded due to some human error. Can you maybe clarify what was that? And would that kind of induce any kind of concern when you think about sort of CNS target down the line. And then second clarification is for your -- the [ B4GALT1 ] program, what's the intended target cell type? Is it hepatocytes? Because I saw myeloid cells on one of your slides. So if it's a naked EON that you're injecting, is it intended to go into your bone marrow? Or is it just in the circulation that it would induce the change.

No. Let me start with the second part of your question. I think the intended target cell -- cells or hepatocytes in which B4GALT1 is expressed. It's not solely dedicated to hepatocytes, but the intended target proteins that we would like to affect are expressed in hepatocytes. So [indiscernible] are expressed there. And through the, let's say, local inactivation or loss of function variant induction, we are going to be affecting glycosylation patents of those proteins and thereby limiting their negative [indiscernible]. So that's the answer to your second part. The first part is we've been working since 7 months to get the proof-of-concept data ready. So we've been gathering information in animal models like mice, wild-type mice but also nonhuman primates. And the first experiment in nonhuman primates that we show actually was intended to dose the 3 animals through intrathecal delivery. Unfortunately, due to human error, 2 of the animals got a faulty dosing. So we didn't deliver the dose. The dose wasn't delivered in appropriate let's say, functional compartment for us to measure the editing. Since the data of the third animal was very robust, and we decided to show it. But in the near future, will be going out with additional points of data in nonhuman primates. So you can rest assured that, that will be a continuous effort to show editing in large animals as well.

The next question comes from Keay Nakae at Chardan.

A couple of questions for sir Gerard. In this application, for your synthetic oligo where you're trying to recruit endogenous ADAR, what factors contribute to the length being 10 to 20 nucleotides as you stated earlier?

No, you mean why can we go so low -- so small?

Yes, how do you know in this application, that's the optimal length.

Well, so the EONs as we develop them are between 25 and 30 nucleotides long. And that's the, let's say, the intended length. We will do a length optimization in each and every target. So there will be some variation to the length. But in general, what we see in our development and our EON designs, we are around 25 to 30 nucleotides, which is which is a length that we can develop as we were -- would be doing for an [ Axon ] skipper or another loyalty mode of action. I think what dictates the length because [ Rene ] referred to at the start with oligonucleotides that have been structures to intend to actually recruit ADAR in the beginning. And by understanding more what ADAR can and cannot tolerate, we were able to buy also chemical modifications to decrease the length. And we are finding that the current length that we are using, which is again between 25 and 30 nucleotides is most optimal to provide a double-stranded yes, let's call it a docking loyalty for ADAR to bind to and then to exert the [ emanation ] reaction on the intended efficacy.

Great. And Dr. Beal talked about how beneficial having the crystal structures has been in the understanding and facilitating a rational design what other technologies or characterizations do you think are needed for you to develop even more efficient EONs.

So indeed, the crystal structures has been discovered by Dr. Beal, we're very, very helpful in modeling the binding of the oligonucleotide to both the target RNA and the ADAR itself. So it allows us to model introduction of chemical modifications at different locations and then the interaction with the amino acids of the ADAR. In addition to that, as we explained, we have optimized the editing enabling region or EER to mimic something that we feel that is most optimal by using a chemical modification. So modeling and understanding the chemical interaction is actually key to develop these programs further. So we are actually expanding the collaboration with Dr. Beal that we also understand that certain sequence contexts, which are less amenable to editing, we can now resolve. And there are some really nice scientific data also being published that shows that we can overcome certain limitations as well.

Okay. And maybe a question on the IP. Obviously, you have a nice portfolio here that you've built. And maybe back at the 10,000-foot level, what is it do you think you own? Obviously, recruiting endogenous has proven itself to be the way to go here. You're using your EONs and the EER being a critical design point. So maybe just recharacterize what do you think you own on the IP front here for ADAR.

We own the world as mentioned. One more thing. If you want to use an ADAR -- and EON to attract ADAR, it's us. we have the patents. We have also confirmation. It's not us telling you that we have a leading IP position, but it's now confirmed also in Europe by winning a very, very important position. So that's the long and the short. You can look it up in our patent slide. Happy to take you through in detail, but the short and the long story is, if you want to use an EON to attract ADAR, it's us.

I'll turn it back to Daniel for concluding remarks.

Yes. Thank you, Sarah, and thank you all for attending the session today. We're pleased to have shared with you the progress we've made on the RNA editing front and outline the plans going forward, and we look forward to keeping you up to date. Thank you all. Have a good day.