 how to work this. OK. Thank you very much. It's important, first of all, I thank Lita and Owen for making sure that I got here. I really appreciate this. I also want to point out that I'm going to talk to you about something completely different, really different. So if you really want lunch, you can leave now. If you are intrigued, then stay, at least for a little while. So it summarized what I'm trying to do with some other people at UBC is we're trying to look into the question of getting biologics from microbiomes. And this is something which is an ongoing project, but it's very interesting and it's very challenging. Having mentioned microbiomes, I have to tell you that my association with the Human Microbiome Project has been one of the most exciting times of my whole life. That's a long time, too. And I feel that the Human Microbiome Project has really revolutionized microbiology. Microbiology is different. We don't do the same kinds of things any longer. We can do so much more. And I think this is probably one of the greatest benefits of the Microbiome Project. And when you get undergraduates coming to you, ask you if they can work on a microbiome, that means something. People know it, they think it's good, and they want to keep on doing it. However, I don't want to talk to you about this, and I can, okay. I will stop by pointing to this side. No. Oh, it's, where do I go on? Oh, it's there. Somebody hit it. Okay, fine. Right, so I'm an antibiotic person and I've been working with antibiotics for a long time. I don't take many antibiotics. In fact, I've taken very few. But I'm interested in antibiotics and especially in the history. And you see, we start this side to this side, you see something about 60 years of the development of antibiotics. In that 60 years, enormous numbers of things have happened. We started with a very small business and we've now reached a stage of a major business. And one that I just want to point out to you, one of the most important things is the time of the FDA office of new drugs. Before that time, there were no clinic, essentially no clinical trials on antibiotics. And most of the antibiotics that we have today are drugs that were discovered before that time. And so they were never really approved, but they were approved, of course, because they were used generally. And so it's difficult now to get antibiotics into the market. So how do we get new antibiotics into the market? And the point is to try to get them in terms of new compounds, novel compounds, which might have a much more rational basis for their discovery. The other thing I want to point out from this slide is concerns the use of antibiotics. And there's been a lot of discussion in this meeting, rightfully so, about the roles of antibiotics in modifying the microbiome or doing other things to people. And I want to point out that in the 1940s, 1950s, there was about 50 kilograms of antibiotics produced per year. And that increased. And the best figure I can get now is that it's a million metric tons of antibiotics are produced at the present time, and probably more because we don't have very good details. So the question is, a million metric tons is an awful lot of a very active compound. This is dispersed into the environment. So anybody who says that they've never had an antibiotic is lying. Because you're bound to have it. You're bound to take it. You can't avoid them. Wastewater treatment plants do not remove antibiotics. They remove some of them, but not many. The point is that antibiotics are pervasive. And not only antibiotics, but people don't think very much about compounds which are used as antimicrobials. Trichocan, for example, is made in hundreds of, no, tens of tons every year and dispersed into the environment. It's a very stable compound. This is something which also will affect the microbiome. And I don't think any studies have been done on this at the present time. If you want to test it by some crest toothpaste, that's all you need. Now, I want to go on to tell you a little bit more about the fact that there was great disenchantment in the antibiotic discovery field in the 80s, simply because they thought that recombinant DNA was going to change the discovery of antibiotics. This actually did not turn out to be the case. But what it did, what has happened in antibiotics is that we've had more and more resistance. Constant use of antibiotics is constant resistance. This is a wonderful example of a resistance gene cluster which is found in Acineta Bacta Balmani, a serious hospital pathogen. And this cluster of genes all are involved with antibiotic resistance to various types of antibiotics, various forms of inactivating, taking the antibiotic up, whatever. The point is that these clusters of genes have evolved over time. I don't think there's any reason to believe that these things existed in the organism before antibiotics. But they are now available commonly to antibiotics, to organisms under any circumstance. So we face a real problem with antibiotic resistance, with the constant dispersal of antibiotics. And so there really is a need for trying new approaches. To finish up a little bit on antibiotic resistance, this reaction is the hydrolysis of a beta-lactam antibiotic, that enzyme which cleaves this ring. That is the most expensive enzyme in history. That is destroyed, well, it isn't expensive actually, it's quite cheap for bacteria, but it's destroyed many, many classes of antibiotics. And we're still trying to produce beta-lactamases. And we're doing a good job, but not good enough. This particular slide shows you the development of beta-lactamases over the period from about 1970 to now. And you see that during this time, a thousand different beta-lactamases have been identified in bacteria. Bacteria are evolving very rapidly. A new beta-lactam antibiotic means a new beta-lactamase. And this is happening all the time. So we can't possibly win. This top of the slide here is interesting because this shows you the evolutionary tree of antibiotics, which is the efforts of the pharmaceutical companies to try to produce compounds that resist beta-lactamases. And you see they're constantly going up. Companies are very creative. The chemists are very good. They can change the structure such that there will be a short-term effect of activity against the beta-lactamase containing strain. But they don't last forever. So we really have to think a lot more about novel approaches to getting new compounds, whether they will be resistant to something, whether they will avoid resistance is not to be taken. It can always happen. But the point is, we need to find more compounds. And the situation now is that these are all of the early antibiotics. And here we have new compounds here. I don't have the very latest one. But the antibiotic industry is totally in disarray. They're not interested. They cannot produce enough compounds any longer. They cannot produce the right compounds. And these compounds at the end here, they're mostly used in animals, which is just as bad, of course. So what can we do about it? Well, thinking about antibiotics, you have to begin to ask the question is, when is an antibiotic not an antibiotic? And you begin to ask the question then the word, where does the word antibiotic come from? And it's really a misnomer. And I'll come to that in a moment. But the point is, the evidence at the moment is that natural products, which we call antibiotics and lots of other compounds that are related to them, are probably acting as signals within bacterial populations. This is their natural role. Their natural role is not that of antibiotics. And in fact, it's nobody, to my knowledge, has ever isolated an antibiotic from soil and shown that it had antibiotic activity. I'm sure it will be done, but it's going to be found in very, very small quantities. Why do I say an antibiotic is not an antibiotic? It depends on the concentration. And this gentleman here, who I fancy very much because he seems to have done everything one could possibly do. And he has one of the most wonderful names I've ever seen. But Paracelsus, who came out with a statement that the dose makes the poison. And this is something that applies really very well to antibiotics because at an appropriate concentration, a poison, a toxin, or anything like this is an antibiotic. So you can shift the concentration and you can go from an antibiotic to a toxin to something which has no activity whatsoever, but accept as a signal. And I think that this is one of the things that we are accepting now that in nature, in natural circumstances, these compounds are being used largely as signals. As I say here, they're rare in nature. So examples of signaling. There are many, many examples of signaling and very unusual examples of signaling of using antibiotics at low concentrations in order to affect other organisms. I don't want to go through all of these, but this is a very good example from Roberto Coulter's lab. This is a strain of bacillus, you see here. It forms no biofilm. However, in the presence of nystatin, which is an antifungal agent produced by a bacterium, at very low concentrations, it forms a biofilm. So that the compounds induce changes, specific modifications in the functionality of the bacteria. And one can see this in many, many cases. Here's another example of several antibiotics, which some of which induce swarming of bacteria, such as this one here. Others that have no effect on swarming. In other cases, we can see that different antibiotics will turn on the production of an antibiotic from another organism by means of diffusion through the argon and activating functions in that organism. There are other cases, and this is an example that we've just recently come up with. I'm very proud of it. This is a gene transfer experiment using an organism called rhodobacter, which has a very unusual gene transfer system. Genes can be transferred very rapidly by a rhodobacter by means of what is called a gene transfer agent. The gene transfer agent activates, will transfer at about, well, this level, as it's shown here, first this. There. But if you do this gene transfer in the presence of a sub-inhibitory concentration of several different antibiotics, you find you get an increase of between 100 and 1,000 times in gene transfer. So that all antibiotics, interestingly, will affect bacteria in different ways. And gene transfer is probably one of the ways we least want to do it. But it happens all the time, and it's because of our use of these compounds. One of the compounds that we've, or class of compounds that we've been working on is the cyclic dipeptides. These are very interesting compounds. They're widely made. They're made by many different organisms. Organic chemists hate them because they don't present any problem in terms of structured determination, and you find them everywhere. They call them nuisance compounds. But cyclic dipeptides have a huge variety of activities, and I've listed some down here, or here, and you can see that many factors that are needed in bacterial populations, many things which lead to bacterial change or the formation of, let's say, common functions within bacteria can be induced by very low concentrations of compounds that are antibiotics, or are they not antibiotics? We don't really know, but I would suggest that they're antibiotics when we use them for clinical reasons. They are not antibiotics if we use them in low concentrations, as they would be in nature. Now, nature. Bacteria, as we know now, bacteria in nature do not grow as single colonies. They are always growing in communities. They grow as mixed communities, and they are held together by a variety of reasons. The communities, they have, in one case, they often have distributed metabolic networks. They share the workload. The fact that they can carry out functions common to a particular pathway means that the organisms can stay together and work together. The other thing is that they can be induced to form combinations of this type by the presence of bioactive small molecules, such as swarming, such as the case of biofilm formation that are induced by specific physiological effects. So in our gut, our gut is not just a bunch of bacteria. It's a very complex community, a biochemically and metabolically related community. And we have to think about it as such. The same is true in skin. Skin is a community. If you affect one member of the community, then you can affect the other members of the community also. In the case of humans, in the GI tract, we produce lots of compounds, many, many different types of small molecules. These small molecules are probably used in many, many cases for signaling between different bacteria, but they may also have other functions. You better put some dark glasses on, right? I just don't wanna forget the people on that side because they're the people that are paying me. So it's important that we realize that the gut is just another community of many microorganisms and these microorganisms are living their own lives and we benefit from them. The fact that they determine our lives is another matter. So we've been thinking about the possibility for several years now of trying to isolate small molecules from gut bacteria, either by isolating the gut bacteria and testing them for small molecules or trying to do it in a batch fashion. And it's proved to be very difficult and I don't want to go into that at the moment, but what I do want to tell you that there is very strong evidence for chemical signaling between animals and their microbiota. And there was a very nice review recently by Margaret McFool Nagai on the interactions between different organisms. Here we go, between different organs and using different types of signals. And as was mentioned earlier, the adrenergic hormones are very common signals in these cases that link organs to microorganisms. And to human cells. The, as was mentioned also this morning, the adrenergic hormones are made by bacteria. And many bacteria actually make a number of different adrenergic hormones. So the fact is that the bacteria have evolved to work with us, they interact with ourselves and they can influence ourselves very, very clearly. It's the use of adrenergic hormones by bacteria has become so common that the field of microbial endocrinology has been proposed. I don't think it's really necessary because we don't really know what the adrenergic hormones do in bacteria, except that some do affect biofilms and some affect other forms of growth. It's very easy to test this kind of thing. But what is this in reality? We don't know because we can't test them in situ. So I want to tell you now that we have to think about a different kind of relationship between the chemicals in nature. The central dogma is DNA, RNA and protein. It doesn't go any further. But I would say that it's, well, not me. Stuart Streiber first proposed this, that natural products were the important endpoint of these three pathways. And we now believe that these products are actually the lexicon of biology. They are involved in signaling throughout all different kinds of organisms and very different kinds of organisms. And this, I believe, is a way in which we can approach the whole question of looking at different organisms and finding ways of studying and controlling different organisms. So finally, where are we gonna get all the compounds we need? Well, it's fun to talk about numbers, especially when it's money that you have. But these numbers are the numbers of bioactive small molecules that you can find in the biosphere. And they're huge. This is, so I don't know if you can see this, but there are four times 10 to the 11th stars in the Milky Way. There are one times 10 to the 24 stars in the universe. And there are probably one times 10 to the 30 bioactive small molecules in the biosphere. These are compounds that we can look at. They're unlimited. We just have to find them. We just have to identify them based on what they can do for us. Now, how do we get them? This is becoming, this has been a very difficult problem. It's getting easier. For years, one isolated compounds from nature by isolating an organism which made a compound, a plant or a bacteria, and grow them up and isolate the compound, purify it, et cetera, et cetera, et cetera. Now things are changing. Microbial genetics, the microbiome is changing this because it is now possible to use genome or metagenome sequences and to scan these genome and metagenome sequences and identify then classes of biosynthetic clusters. Once you have a biosynthetic cluster, it's possible to clone that cluster into a high producing organism and identify the product. So this is a case where you start with an organism and you look for a biosynthetic pathway. You don't know what it is, you don't know what the product is that is being made, but it's possible to make compounds which are previously unknown. In the same way, one can use the technique of scanning math spectroscopy, which is a very exciting technique which has only been used in the past few years by Peter Dorisstein to look at the presence of small molecules, identify small molecules, and made by bacteria in interacting with other microorganisms. And this is shown here. There are, you can notice that there are many small molecules which are identified by these clusters and the system is so sensitive that one can get an accurate identification of the compounds by math spectroscopy directly. This is gonna change the way we carry out drug discovery. There's no question about this because it's now going to be possible either to look for a compound and then look for a biosynthetic pathway from that compound because once you know a compound, you can identify a biosynthetic pathway or you can try to find a biosynthetic pathway first, as was mentioned in the previous talk, finding a pathway for a polyketide synthase is relatively easy. And now it's relatively easy to make the compound which is encoded by that pathway. It was not always so easy. And I think the human microbiome project has helped enormously in leading to this stage in natural product drug discovery. So I'm gonna close now, this is my lab. We've got three people working on microbiomes, not the microbiomes that you might be interested in, but we do it and it's fun. And finally, I would like to point out that the gaps and challenges that I see in this particular field, first of all, the state of the art mass spectroscopy is changing. It's being developed all the time but it's also incredibly expensive. So I suggest all of you write to NIH and ask for a mass spec soon before they go up in price again because they're good now, they're not the ultimate, but the point is that they will be things that you can use to identify small molecules. The other thing is the isolation of active compounds. This is not always easy, but it's getting better. Again, one needs good software, one needs mostly skilled chemists. And then finally, I'd like to point out that we're having this meeting as the end of the NIH Human Microbiome Project, the big one, but the human microbiome is not finished. If we've seen here at this meeting, there's still many, many things that we have to learn about it and I do hope that funding is given to continue this. You know, I'm doing this entirely in a charitable fashion because in Canada you can't get money for this kind of thing but you can, so do it and invite me to help you. Actually, I haven't finished. I've got one more slide and there's something I want you to do before you have your lunch. Thank you, Julian. We have time for a few questions but before we do that, I just want to urge you to stay around after the session. Leader Proctor wants to make a few comments so don't rush off to lunch. Do we have anybody queued up? Well, I love your global overview, it's absolutely universal, it's terrific. And your play with numbers, is that astounding? I wrote as many down as I could, I hope we get some of your slides when you're finished. They're stunning. But I was really intrigued by the period of demoralization by the drug companies when they simply couldn't continue to develop new antibiotics because resistance was developing even faster. So now that you've come up with this small molecule strategy, are the drug companies becoming more enthusiastic or are they still in a state of depression about antibiotics? Yeah, the drug companies are becoming more enthusiastic. They're not investing a large amount into it at the present time but there are a number of small companies that are doing this and they're doing it much more vigorously. And if anybody is going to succeed in finding novel compounds that will be useful in therapeutics, it's going to be small companies now. I apologize to any big farmer people here but I'm sorry you don't have it. Thank you. So, Julian, do you have any thoughts about the potential for adapting some screens? A lot of screens are using reporters and my thought is that typically these are done with a limited number of reporters but with tractable organisms, you could potentially make a lot of reporters and sort of do combinatorial screens with many, many different reporters and many, many different compounds kind of in the way that chemists do combinatorial chemistry. Yeah, I think reporter screens are going to be probably the most important way to get compounds and I'd be happy to talk to you about it and if you can tell me your ideas, I'll work on them. That'll be fine. Thank you. Okay. Okay, let's thank Julian for an excellent talk. Okay. And I'll turn the session over to Lita.