Thank you, Nigel. And thank you very much for joining us here at this event. It's my privilege to share with you some more recent developments, especially around NNH [sp] transformative research program, which is called Mapping the Human Toxome, which I have the privilege to direct.

Nigel has already kindly introduced my background. I'm a toxicologist and pharmacologist interested in novel testing approaches. And these novel--the motivation for doing so is multiple. First of all, we all area aware toxicology is an important business. It is not only consuming roughly 3 billion of testing cost for animals, but it's more importantly regulating products at a trade worldwide of 10 trillion with essentially the same set of methodologies.

But, we are increasingly aware of some problems associated with these traditional methods, which have in large part been designed some 40 to 80 years ago. We have a problem of throughput. We have to deal with about 100,000 substances and consumer products, out of which only roughly 3 percent have been really extensively tested.

We have a cost problem with individual assays costing sometimes more than 1 million. If you think about development in neotox or cancer, we have a productivity issue. We are not 70-kilogram rats. We are understanding increasingly that the methods are making the world possibly safer for rats, but not necessarily for humans.

We're seeing that we have developed the very precautionary type of toxicology, which might make it difficult to bring something like aspirin to the market today. We're seeing that animal use is considered more and more critical by a general public. And we are seeing that new products, like biologicals, nanoparticles, cell therapies, are requesting different type of approaches for safety assessments.

We're also facing permanently new hazards of concern. For example, when nanoparticles are now possibly being associated with effects on arthrosclerosis, this was not a classical toxicological endpoint we have been looking for.

And our repertoire, our toolbox, is not adequate for addressing mixtures or the problem of individuals in toxicology.

So, the advent of the report of the National Academy of Sciences on toxicity testing in the 21st century in 2007 has brought about I think an atmosphere of departure. And we can witness this in every year's program here at SOT more and more sessions being devoted to how could we do better.

And the key messages of this report were, from my point of view, first of all to embrace new technologies. It is calling to make use of the revolution in biotechnology and in bioinformatics of the recent decades. And it is suggesting to go for pathways. It is suggesting to make our signs more mechanistic, even to the level of molecular definition.

And this is impacting. And just to give you a few, let's say, highlights to show how much of a mind--change in mindset is taking place, for instance, Collins first authored a paper in Science only one year later writing, "We propose a shift from primarily in vivo animal studies to in vitro assays, in vivo assays with lower organisms, and computational modeling for toxicity assessments."

In a similar tone, Peggy Hamburg, Administrator of FDA, last year in an editorial for Science, "With an advanced field of regulatory science, new tools, including functional genomics, proteomics, metabolomics, and throughput screening and systems biology, we can replace current toxicology assays with tests that incorporate the mechanistic underpinnings of disease and of underlying toxic side effects."

And Jackson, the Administrator of EPA is even a step further. Already in 2009, they fully embraced the report of the National Academies of Science and made this EPA's chemical toxicity testing paradigm.

So, we're seeing here from various agencies movement towards change into a toxicology for the 21st century.

So, what is the concept behind this? There's 50 ways to leave your lover, and there's probably a couple of hundred ways to harm a cell. But, we believe it is not an endless number. For the very simple reason that the number of critical infrastructures in a cell, which need to be harmed to lead to a problem, is limited. And if so, certain pathways must converge in order to harm these pathways.

It is least an hypothesis which is put forward. And the idea is that, if we would be able to annotate these pathways, to describe them on a molecular level, we might be able to come up with something like the human toxome as the entirety of pathways of toxicity.

And we could start annotating these pathways to cells, to different cell types, and understand why a hepatocyte is vulnerable to a toxicant while a cardiomyocyte is not. We could annotate them to classes of toxicants, understanding what are the critical perturbations which lead to a certain phenotype or has a manifestation.

And this annotation could take place also for a species even, understanding why a certain species is vulnerable, another one is not, because a certain perturbation can place--take place or not.

And there's a certain beauty in this because, if we have such a comprehensive list and understanding what are the crucial pathways for a certain manifestation, we might for the first time be able to actually say that a substance is negative.

So far, the next animal experiment can show us there is a problem which we have overseen. So far, changing any condition in a cell system might come to a different result.

But, if we know that there is no perturbation of a canonical pathway necessary for a manifestation of hazard, we could actually say this substance is clean. Here, we can close the books.

This is a slide coming from the Academy of Sciences in their presentation of the report. And this is examples they have given us on the way--what pathways of toxicity could look like.

And if you see that so different things like endogenous hormones, DNA damage, or hypo-osmolarity among them, you might understand that it is quite a challenge to find a common annotation, a common nomenclature for annotating such type of pathways.

Nevertheless, we have teamed up between a group of people in order to form a partnership start to doing exactly this, trying to develop an annotation for pathways of toxicity and a public database to govern this.

And we're trying to team up with technology providers like our hosts, the industries interested in this work, and experts in the field. And we're trying to bring in the various technologies which lend themselves to do--for doing this, mainly transcriptomics and metabolomics at this stage.

We need conceptual work in order to make these things happen. We need the failures of the past and the experimental models, which have been used to resolve these cases. And we need expertise to identify the pathways of out of these omics data.

And we have been granted an NIH transformative research grant, which started working in September last year, which is entitled Mapping the Human Toxome but Systems Toxicology. And this very humble first effort is only addressing one single hazard, which is estrogenic endocrine disruption. It is only addressing two cell lines and a handful of toxicants because the goal is not to develop the next endocrine disruptor test.

But, it is the goal in an area where we have quite a few--quite a bit of understanding of the pathway character of this toxicity to develop the concepts of annotations and the respective database and how to pinpoint these type of pathways.

This has been complemented with FDA funding to our laboratory for identifying pathways of development and new toxicity so that we have a second type of hazard where we try to explore the respective opportunities.

And we're trying to form on this basis now PoToMaC, the Pathway of Toxicity Mapping Center, so an entity to create a workflow of systematically analyzing samples with these technologies and trying to deduce the pathways of toxicity behind them.

So, our starting point is actually using validated or pre-validated cell systems. You can imagine from my background as the former Head of the European Center for the Validation of Alternative Methods, I'm very much aware of the needs of quality control and standardization and the need of having predictive types of cell systems.

So, these systems are characterized by robust protocols, which have undergone evaluations in international ring trials. They're good sell models. They have found to some extent regulatory acceptance. So, we believe that pathways derived from such systems will actually be more acceptable because we explain why these models have given us correct answers.

They're also characterized by the availability of reference substances because we know which set of substance has been correctly identified in these test systems. And even more importantly, we know the thresholds of adversity.

While a normal in vitro system, which we publish on, has no thresholds, has not defined outcomes, an alternative method needs to have this in order to predict animal or human type of reference values.

We know at which concentration a certain substance is called a positive. So, we are able now to understand which pathways are engaged at around the levels of adversity.

And in the end, we are taking advantage of about $300 million of investment, mainly in Europe, into the development of these validated methodologies.

As I said, in our NIH program, we are addressing proestrogenic endocrine disruption, especially because this is a very much pathway-based toxicity, which is reasonably well understood. We also have a variety of other tests available, which we can use for comparison to understand how canonical certain pathways are. And obviously, with the endocrine disrupter screening programs, there is a testing need so that there might be windfall benefits from our work.

We also have a very nice set of reference substance, which has been defined in the equine [sp] process. And we're employing metabolomics and transcriptomics and their integrated use by informatics.

And we are especially happy to have Agilent as a partner here, bringing in the emerging bioinformatic integration for pathway identification, which has been presented to you.

We are well aware that there is quite a few of challenges in executing this, as we have no concept yet for what is a pathway of toxicity. There is no common definition which is accepted broadly. We don't have mapped a single pathway yet successfully. The annotation, the language to describe this on a molecular level has not really been done. And nobody has come forward with ideas how to validate a pathway of toxicity once it has been identified.

We also might encounter problems of temporal and spatial resolutions as these pathways are not necessarily linear, both in respect to time and space. We also will have to figure out what is pathway and what is a variant of a pathway. And when are two variants distinct pathways?

We'll have to start defining adversity from pathways. So, when is it actually harmful that a pathway has been disruptive and when not?

No database exists. And we have no idea how the governance of such a database should look like. And we will certainly need more technologies than just transcriptomics and metabolomics to achieve the ambitious goal we have set.

If you want some overview of this project, this is last week's issue of Science, which is also available here at the AAAS booth, which was reporting on these activities and in this respective article. So, this is a very nice short summary.

But, there's also much more discussion and conceptual work around this. And this is a series of articles which we have published in ALTEX, an international journal, which we have--which we are promoting and using. We are running the U.S. editorial office.

This journal has in the meantime an impact factor of 4.4 among others because of the conceptual type of articles which have been published and are quite nicely cited. This is some examples.

And as a preprint of the next issue, the one marked here in blue, [unintelligible] and Systems Toxicology, which will be published the next issue in May, is already available here at this--on this occasion and at the Agilent booth.

It is one example of the discourse which we are trying to promote in order to develop the necessary concepts for this new science. So, what is this about? And this is a 10,000-foot perspective. In the end, it is trying to move from an empiric type of view to a mechanistic type of view.

We have identified very empirically in the past predictive models. And we know the reference substances which are associated with this. Let me see. Is this--no, there's no pointer. Is there not?

So, the--and we have reference toxicants which work in these models. Omics phenotyping allows us now to derive signatures of toxicity, which means we identify those genes which are typically altered in their transcript, in their transcription or the metabolites which are changed due to the effect of these reference toxicants in our predictive cell models.

And this will help us to derive the pathways of toxicity which are leading to this which are forming the toxome and moving from a purely correlative and empirical type of approach to a pathway of toxicity approach.

And we would hope that this at a later stage allows us to start modeling these exercise--these different pathways into a dynamic systems toxicology type of methodology.

So, the--we're at the moment at the stage of omics phenotyping towards signatures of toxicology and trying to derive the pathways of toxicity. But, as you can see, the overall goal would be to develop mathematical models which allow us to make predictions out of pathway disruptions.

And in the identification of these pathways does not only feed the signatures of toxicology, of toxicity, which we're identifying in this unsupervised way, but also our mechanistic understanding, which we have derived over the last few decades in molecular toxicology and mode of action type of analysis.

And last but not least, we can draw on the wealth of information from biochemistry, from molecular biology, which is giving us increasingly pathway information, as does molecular medicine, trying to help understand which pathways are crucial if they are disturbed to form a certain type of manifestation of hazard.

This is a slide which is borrowing from Hans Selye, well--world famous when he described in the '50s already systems under stress. And he made a point with a very similar slide. And I actually only changed the wording on his very own curve, which is that a system which comes under stress moves from homeostasis. It is disrupted. And some type of counter-regulation allows it after awhile to establish a new type of homeostasis.

And only if the exposure to this stressor continues or is overwhelming, we see an exhaustion. We see a manifestation of problems at the end. But, a system would always try to cope with stress by altering--by reaching a new way of homeostasis.

And in this slide, I tried to put in, first of all, the chemical biological interaction, which is resulting in a pathway of toxicity. The pathways of defense, which then bring the system back into the normal direction, quite a few of epiphenomena, which are then in the ultimately leading into a certain type of new homeostasis.

And it is only the manifestation which takes place if time and dose together are overwhelming the system.

So, what we are interested in is the pathways of toxicity, the chemical biological interaction, and the mechanisms which are set in motion in order to harm the system. What we're, however, typically measuring is over here. We're measuring at sub-cytotoxic concentrations. But, we're measuring at an established new homeostasis. And this is obviously one of the problems we have to overcome if we now want to understand how the system is actually rearranged.

We're trying to establish a workflow for toxicometabolomics, trying to go from sample preparation to measurement in this cutoff [sp] LCMS, for example, analyzing the features which are detectible using principle component analysis to identify those which are correlating with the toxic or nontoxic character or the dose response relationship of the given toxicant and coming from there to the identification of metabolites, leading hopefully in the end to the identification of pathways of toxicity.

And we need some technologies, cell biology, mass spectrometry, bioinformatics, to do so. There's quite a few of challenges associated with this. There's a certain complex detection method in the biological part. There's needs for standardization. The sample handling is not yet standardized. There's reproducibility issues. We need databases to identify the metabolites. We have throughput limitations. And there is multidisciplinary expertise required to actually identify the pathways of toxicity.

In order to develop these concepts, we have started a series of workshops in the context of a trans-Atlantic think tank for toxicology, which we entertained together Buzz Blauber [sp] at the University of Utrecht and Marcel Leist at the University of Konstanz in Germany.

This think tank is sponsored by the Doerenkamp-Zbinden Foundation. And in a series of workshops, several associated with topics relevant to this project, we have just carried out in February jointly with BSF a workshop on metabolomics, which brought together several groups using metabolomics and toxicology. And we're planning for a series of workshops nw on identification of pathways, metabolomics, and integration of different omics technologies in the course of this year. And we hope to develop some of these concepts further.

So, you could say what our task is that we want to actually come from a dinosaur footprint to back to the dinosaur. The Dinosaur footprint is the impact a chemical has left into a biological system, what we can measure. We see the changed physiology. We see the alterations of metabolites and gene expression. And our challenge is now to go this important step backward.

The advantage we have is that we can actually make our dinosaur step again and again because that's the beauty of toxicology compared to other life sciences. We have the disease agent in hand. We can experimentally create disease. We don't have to compare patients to non-disease individuals.

We can also observe while doing so. So, we can observe early events after the chemical has been added to the system. And we can compare to other animals, which means we can compare to other endocrine disruptors or other developmental neurotoxicants in our system to understand what is the difference in outcome with these footprints.

And we have more information. We have bones which are, for example, the knowledge on molecular biology and biochemistry, which is linking the various metabolites so that we have information which helps us to understand what has actually happened. So, we're not just bound to the footprint, the measurement of deranged metabolites itself.

The rough work plan of our NIH project, which involves a variety of groups, which I mentioned in this context, several people involved in the panel of toxicology in the 21st century are involved, Kim Bucklehide [sp] at Brown, Jim Jaeger [sp] at Johns Hopkins, Marty Stevens [sp] with us now at Johns Hopkins, and Al Fonas [sp] in Georgetown University are producing high-quality data.

The pathways of toxicity concept, as I said, is developed in the context of our think tank. Mal Anderson , also a member of the Tox 21 panel at the Hamna Institute, helps us with the analysis based on known pathways of toxicity, EPA tox cast , Bob Cavlock , Imran Shah , and his team are involved as a partner in this project, help us with analyzing, integrating this with the tox cast suite of assays.

And we will do unsupervised pathway of toxicity identification by using the integrated biology approach of Agilent, another partner in our project.

At later stages, we aim for validating pathways of toxicity. We will refine our tools and processes. And ultimately, we hope to establish a toxome database, which allows us to actually create a wiki type of process where various groups can feed in.

We have integrated--or, this--these activities are integrated in larger activities in trying to develop roadmaps for new methodologies of toxicology. And over the course of last year, we have had a very exciting process, which was started by the stock taking on alternative approaches available for cosmetic testing in the E.U. The European Commission had requested a report published in May 2011 by Atlet Al in Archives of Toxicology, which did demonstrate what is the state of the art of methodologies available.

And we have been carrying out an independent reassessment of this report, which added a couple of opportunities but altogether did confirm the tremendous gap we are seeing with regards to alternative methods available.

And we have then carried out an exercise in devising a roadmap how to move forward. And this roadmap was based on five white papers and a consensus workshop with 35 participants. And its result has now been published in this month's issue of ALTEX.

This article, A Roadmap for the Development of Alternative Non-Animal Methods for Systemic Toxicity Testing, is available free for you as a PDF from the ALTEX or the CAT [sp] Website but also at various booths here at the SOT [sp]. We have print copies of ALTEX available if you're interested in this report.

And as a next step, we will, on the 20th and 21st of March, in a multi-stakeholder event, present this roadmap in Brussels in Europe, in Belgium. We'll present these concepts in order to discuss them and to hopefully see that this is impacting on some of the funding initiatives of the European Commission over the next few years in order to make some of these things happen.

Another initiative I think which is the sparring partner of all of these developments is evidence-based toxicology. Some of you might have witnessed last year's SOT, which gave us the kickoff of the evidence-based toxicology collaboration, a collaboration which tries to pursue the quality assurance of evidence-based medicine in toxicology.

This was based on the paper in 2006 and an original conference in 2007 in Como, Italy. And our steering group here, once again, from last year's SOT kicked off a group of people now interested in moving this concept further in toxicology.

Marty Stevens, who is with us here, is the Director of the Secretariat, which we have--which we are holding at CAT in Europe.

The first forum of this event--of this stakeholder group was held--hosted by EPA on the 24th and 25th of January. And the proceeding are in making.

The European branch will be kicked off on the 17th of Juen in Stockholm as a satellite to Eurotox. So, we will make this a trans-Atlantic event as well.

So, taken together, you have seen there's technological developments taking place at the moment, moving us towards mapping the human toxome, try to define molecular pathways. At the same time, there is some type of political programs trying to acquire resources for making this program a real at least trans-Atlantic type of effort.

And we are thinking already now while producing the technologies of a sparring partner of quality control which is the evidence-based toxicology movement.

So, I'd like to close with a quote from John Maynard Keynes, who said, "The difficulty lies not in the new ideas but in escaping from the old ones." And I hope I've shown you quite a few new ideas which are in the making. Thanks a lot.