Attending the HGV2014 conference — Not a sinking feeling

It was my first visit to the Emerald Isle, and most definitely worth it. However short, my three-day permanence in Belfast to attend The 15th International Meeting on Human Genome Variation and Complex Genome Analysis (HGV2014) was fruitful and thoroughly enjoyable.

I flew from London Gatwick to Belfast International Airport on an early morning flight on September 17th, never a good start of the day; not for me at least, given my long-term experience with low-cost flights that leave REALLY early in the morning bound for my native Sicily from London, the city I have emigrated to, and where Nature Protocols is based. You’ve got to suffer to go Sicily, it seems — and be mildly uncomfortable to go to Ireland.

Well, the discomfort did not last long. As soon as I climbed onto a cab just outside Belfast Airport, the friendly cabbie exclaimed “Culloden? It’s the best hotel in Northern Ireland, sir. Five stars, you know?” Culloden Estate and Spa was to double as the venue of the conference and my hotel accommodation. That review by an undoubtedly knowledgeable local sure felt nice to hear. Time to relax on the back seat of the car and take in the scenery on that drizzly morning as the cab drove for about 35 minutes through the Irish countryside, skirting downtown Belfast, before arriving at Holywood, the Belfast suburb where Culloden Estate and Spa is located. The castle-resembling hotel is immersed in quite spectacular greenery, both within and without the hotel grounds, and the hilly locale overlooks Belfast Lough, the deep inlet of the Irish Sea at the end of which is located Belfast, with its harbor. When the cab pulls over by the main entrance of the Culloden, a man in impeccable grey tail-coat and top hat opens the car door for me with a smile. Wow, that’s not the kind of treatment the average former chemistry researcher is accustomed to…

Time to check in, undo my suitcase in my room, and walk to the event room to register for the conference. There, I am handed my name badge “Baldo Lucchese – McMillan Publishers” by Sinead Lawlor, the über-efficient overseer of the conference organization. She explains to me that the conference will be attended by 126 participants, ten of them exhibitors (I think I am included among those ten), and there will be 54 research poster presentations. A pretty manageable conference size-wise from my standpoint. Among the participants are also three researchers who have written protocols for our journal, Dr. Ido Amit of the Weizmann Institute, the corresponding author of High-throughput chromatin immunoprecipitation for genome-wide mapping of in vivo protein-DNA interaction and epigenetic states, and doctors Yves Moreau and Thierry Voet of the University of Leuven, co-authors of Microarray analysis of copy number variation in single cells. Always nice to find one’s footing in that sense at a conference.

Time to dive into the talks. We are welcomed to HGV2014 by this year’s conference organizer, Dr. Mark Lawler of Queen’s University Belfast, who has also been instrumental to my attendance. As he will explain to me the next day, he moved from Dublin’s Trinity College — Mark is actually a native Dubliner (don’t they say “Dub”?) — to Belfast’s Queen’s University barely a year and a half ago, so it has been quite the struggle for him to organize HGV2014 in his new hometown. You surely wouldn’t have guessed from the results, however. I couldn’t be happier with my accommodation, the conference will progress smoothly and the talks will prove excellent.

HGV2014 is articulated into seven sessions distributed over three days, which will include a total of 36 talks and three round-table discussions. Of the 33 speakers from academia and the three from industry, the vast majority hail from either the British Isles or the US, although there are also a total of eight speakers from Canada, Italy, Belgium, Germany, and Israel.

The first of talk of the first session, “Interpreting the Human Variome”, is given by Dr. Pui-Yan Kwok of the University of California San Francisco, a member of the organizing committee and a veteran organizer of this conference. The talk, “Structural Variation in the Human Genome”, seizes my technique-oriented, Nature-Protocolly (warped?) mind. One of Dr. Kwok’s group research focuses is the study of structural variations of the human genome, variations like gene deletions, insertions, duplications, inversions and translocations. Many of these features are associated with particular diseases and disease phenotypes, but they are often hard to identify and define using whole-genome analyses, which rely on the mapping on short DNA reads ‘chopped’ from the full length genomic DNA and matched to a reference genome. This approach can provide ambiguous information when it comes to identifying, for instance, the number of times a gene is repeated in sequence along a stretch of genomic DNA, a rather important parameter to measure in some research and clinical contexts.

Dr. Kwok’s laboratory, in collaboration with BioNano Genomics Inc., a company based in San Diego, California, has developed “Genome Mapping in Nanochannels”, a technique that is particularly suited to identifying structural variations in the genome. In this approach, instead of being reduced in size, long stretches of human DNA are first nicked at several positions on only one DNA strand using a nicking endonuclease expected to nick about 12 times every 100,000 bases. These nicks are then repaired using fluorescent nucleotides and the full-length DNA molecules are then uncoiled and forced to pass through narrow parallel nanochannels, in which only one DNA molecule can fit at a time. The DNA molecules are then photographed as they move along the nanochannels. Given that the nucleotide sequence recognized by the nicking enzyme is known, that a reference genome exists, and that one obtains clear images of where the fluorescent nucleotides have been inserted along the genomic DNA stretch, a researcher can now, for instance, literally count on a photograph the number of repeats of a particular gene. Using this approach, Pui-Yan Kwok’s research group believe to have closed some of the sequence gaps in the human genome, its ‘grey areas’ of ambiguous sequence assignment that are probably due to the presence of structural variations.

The following Session II, “Tracing and Targeting the Tractable Cancer Genome”, was in principle a bit too focused on clinical research from the standpoint of Nature Protocols remit; nevertheless, it did rouse my curiosity. The lecture by Dr. Lillian Siu of the University of Toronto was particularly thought-provoking from my layman’s point of view. Lillian Siu’s research focuses on the study of inter-patient and intra-tumor genomic heterogeneity in the context of cancer. It has been known for years that cancer patients with specific genetic signatures tend to respond (or not respond) to specific drug treatments in a somewhat predictable fashion. Such genetic heterogeneity is the basis of the so-called personalized medicine approach to treatment. Dr. Siu sketched for the audience the clinical trial designs that take into account genetic heterogeneity. At one end is the ‘umbrella trial’, in which all participants share the same tumor type (for instance lung cancer), according to histology results, and they are assigned to different drug treatments based on their relevant genetic aberrations. At the other end is the ‘basket trial’, in which patients with different cancers based on histology are pooled together into treatment groups based only on their genetic aberrations, irrespective of histological evidence. Finally, she described the N-of-1 trial design, which consists of a comparison of the results of two different treatments applied to the same patient: in this trial design, time-to-progression in a patient on a genotype-matched treatment is compared to the same parameter in the same patient when he/she was on a prior, and presumably non-genotype-matched, treatment.

At the end of this talk, a calculatedly provocative question from a member of the audience rang: “So, are randomized, controlled trials a thing of the past?” Well, given my background as former associate editor at Nature Reviews Nephrology and Nature Reviews Gastroenterology and Hepatology, this question made me sit up: aren’t they the gold standard for medical trials? Lillian Siu clearly did not want to be drawn into sweeping generalizations, but she explained that, in a genotype-matched drug clinical trial, tumor responses may be sufficiently significant and objective for conditional drug approval of the drug to be granted in some cases. In this context, she thought that, prospectively, randomized trials are likely to become less and less prevalent, given how exhausting they can be for research resources as well as for patients. It sure was a treat to witness this exchange…

The next day’s morning session was entitled “Phenomes, Genomes and Archaeomes: Solving the Conundrums”. I have had a small obsession for studies on ancient DNA genomics for a while, so it came to no surprise to me that I found the talk “Ancient Population Genomics: Do It All or Not At All” by Dr. Dan Bradley of Trinity College Dublin, particularly exciting. As Dan Bradley reported, a key difficulty when working with ancient DNA is that when collecting DNA from a typical ancient human bone sample, only about 1% of it is DNA from the original owner of the bone, the rest is leftover from various contaminations. He and his research group have used bioinformatics tools that enable researchers to sift through the sequencing data from ancient DNA that does not need to be particularly pure to infer which sequences pertain the individual to whom the biological sample belonged and which are, instead, due to contaminants.

Dan Bradley reported results from his group and collaborator’s research that help shed some light onto the population and cultural changes that Europe underwent between the Mesolithic through to the Iron age, via the Neolithic and Bronze age. One of the overarching questions his group has been trying to address is whether age-defining technological/cultural innovations were exchanged ‘osmotically’ between populations or whether such dramatic changes, for instance from a hunter-gatherer culture to an agricultural one, are associated with new human populations displacing the previously resident ones. Although their data on this subject appears to be a bit of a mixed bag, evidence seems to indicate that in most cases an age-defining technological or cultural shift also coincides with a population replacing or pushing aside another. Well, it does make sense in a sad sort of way.

Session IV “Answering the Global Genomics Challenge — Time to Step up to the Plate”, late morning on September 18th included talks that focused on ongoing international efforts to sharing genetic data across research centers and across countries to share evidence about genetic mutations and how they may be associated with the onset of, or the predisposition to, disease. Dr. Anthony Brookes of the University of Leicester, among other projects, recounted the design and purpose of Cafe Variome project. This project in particular stuck to my mind because the previous day I had had the chance to talk about it with Dr. Owen Lancaster from Brookes’ lab, who was presenting a poster describing Cafe Variome.

As Owen also clarified for me, Cafe Variome tries to shift the basic nature of the problem of accessing data on mutations and phenotypes that, despite their high potential usefulness, are currently not being openly shared due to legal, ethical or competitive reasons. Cafe Variome tries to clear this roadblock by providing a platform whereby the existence rather than the substance of the data is made accessible to subscribers. Once these discovery ‘hits’ are achieved, the platform facilitates exchange of the data in the form the most suits the relationship between data requestor and data owner: data summaries, data displays, data links, data owner contact details, or a form for data automated data request and subsequent provision.

Presently Cafe Variome, which is hosted by the University of Leicester, supports networks of rare disease diagnostic laboratories or research consortia that share an interest in certain causative genes or diseases, know and trust each other to different degrees, and wish to have a full picture of what records and information exists across the collaborating network. Six such federations are currently trialing the platform, and, Owen explained, one of them will be shortly opening up their content also for public discovery. Well, fascinating stuff for someone like myself who is a bit obsessed with data sharing and privacy, particularly when it comes to genetic make-up. Sooner or later I should write a post on that too…

Session V, “Improving our Health: Time to Get Personal”, was particularly engaging. In the first talk, “Personalised Cancer Medicine: Are We There Yet?”, Mark Lawler described the efforts and purpose of the European Alliance for Personalised Medicine, of which he is a member, an international association that aims to promote personalized medicine and its delivery in the health systems of European countries and to develop patient-centered European translational research platforms. It was clear how much enthusiasm and personal stake Mark has in promoting personalized medicine in the context of cancer treatment, but from the last talk of the session it was also clear, at least to me, that he does not take himself overly seriously, and he is quite open to discussion.

The last ‘proper’ talk of this session, aside from being quite entertaining, was rather unusual in that it was given by Professor Timothy Caulfield, a researcher in the area of health law, biotechnology policy and bioethics. The title of talk, “Marketing the Myth of Personalized Prevention in the Age of Genomics”, already conveys a lot of the tone of this presentation. Timothy Caulfield is a bit of a skeptic of personalized medicine, and he started his lecture by thanking Mark Lawler for inviting him to talk at the conference despite his stance and, presumably, he implied, as a way to kick-start a lively discussion. Caulfield does not believe that the prevention of common diseases is likely to be affected by personalized medicine. Progress in genetic research, he argues, has been hailed as heralding a ‘revolution’ for decades now. But this revolution has yet to materialize. He is also rather offended by what he calls ‘science-ploitation’, via which for-profit companies, even those commercializing beauty creams, for instance, claim that their products can be targeted and tailor-made to suit the personal needs of customers based on their genetic make-up. A number of sleek commercial ads with portentous ‘scientific’ claims succeeded each other on the screen at various times as Caulfield gave his presentation.

The core of his objecting to the promotion of, for instance, widespread genetic testing in the healthy population in the wake of the personalized medicine ‘revolution’ lies in the research that points to the fact being made aware of a predisposition to a particular condition or disease does not change the behavior of individuals to any significant or measurable level. Focus on genetic testing and personalized approach when it comes to public health policies, he argues, shifts responsibility from the political and social level to the individual, when it is proven that only ‘general’ policy decisions can spur changes in social attitudes and, ultimately, change the behavior of individuals. To drive home the point about science-ploitation of the concept of personalized medicine, Caulfield recounted how he had had his genome sequenced by an American commercial company. Caulfield shared with the audience some of the results he had obtained, which meant that he had a slightly elevated predisposition, compared to the average, to a few medical conditions. The ‘bespoke’ advice that he received from the commercial company was to exercise, drink in moderation and not smoke… So much for the personalized prevention regime; point made, I’d say. Science-ploitation… That’s a word that I’ll end up using a lot, I’m guessing, sorry Professor Caulfield.

This session was capped by a round table discussion entitled “Prime Time for Personalised Medicine”, which had a TV talk-show format, in which the chair, conference co-organizer Dr. Stephen Chanock of the National Cancer Institute, Maryland, moderated the discussion of a five-membered panel that included also Timothy Caulfield, Mark Lawler, and Lillian Siu. Of course different points of view and opinions were shared during the discussion, but the panel ultimately agreed that the sensationalization and mis-representation by the general interest and popular press and by commercial companies of the results and implications of genetic research are a significant problem for the scientific community. However understandable sometimes, these tendencies could ultimately harm genetic and genomic research itself. A tentative solution proposed by several of the panelists and a few members of the audience was for scientists to become more actively involved in the public discourse about genomic and genetic research, so as to offer additional means for the public to correctly frame this discussion and interpret the significance of research findings. Proposals were made that scientists should try, for instance, to make themselves more savvy and effective in the use of social media like Twitter, although some confusion immediately ensued this proposal as some panelists and members of the audience couldn’t initially agree on the character limit of twitter feeds. Nevertheless, it was an encouraging start all in all, given that the matter was eventually correctly settled: that limit is 140.

An excellent conclusion of the day’s lectures, before the gala dinner of the conference, which took place at the Titanic Signature Building, an attraction that opened to the public in 2012 and is located in the section of Belfast harbor where the RMS Titanic was built. This building, which is exactly as tall as the Titanic’s highest tip, hosts a multi-storey, multimedia museum themed on the building and ultimate sinking of the Titanic. Us conference participants were taken on a guided tour of the museum that chronologically followed the Titanic (mis)adventure and that culminated in a viewing of the striking underwater footage of the wreck of the Titanic shot from a deep-sea submersible in the 2004 expedition undertaken by marine explorer Robert Ballard together with the National Oceanic and Atmospheric Administration (NOAA). Naturally, a very fancy and very satisfying dinner followed, made particularly enjoyable by light conversations with my table mates, researchers from Spain, the Philippines via California, Britain, and Sweden.

The last day of the conference was dedicated to sessions VI, “Understanding the Evolving Genome”, and VII, “Next-Gen ‘Omics and the Actioanable Genome”. It was also the day of two Sponsor Presentations, one of which was by BioNano Genomics Inc.’s Dr. Han Cao, “Towards the True Contiguity Resolving the “Dark Matter” of Genome”, which covered also the technique already introduced on the first day of the conference by Dr. Pui-Yan Kwok for Genome Mapping in Nanochannels, and that BioNano is currently applying also to the study of non-human genomes. As mentioned, pretty fascinating stuff from the point of view of a technique-biased scientific reporter.

Talking about techniques, the last day of HGV2014 was also dedicated to giving awards to the best posters. In advance of my attendance of the conference, Nature Protocols had proposed to award a one-year online personal subscription to the journal to the presenter of the poster describing the most interesting technique, and Mark Lawler had been immediately enthusiastic and supportive of the idea. As I walked around speaking with poster presenters, I also got to chat with Nature Protocols authors Ido Amit and Thierry Voet. They were possibly being nice, but they both told me that they enjoyed their experience as authors of our journal, and they considered Nature Protocols a very useful resource. It may not be statistically significant with just two subjects, but they both ended up putting more work than they had anticipated into their Protocols, but they felt it was entirely worth it. A quiet sigh of relief on my part welcomed the conclusion of their reflection. Surprisingly, both of my interlocutors, to whom I spoke separately, by the way, also felt that it would be of great help if Protocol authors could submit video tutorials alongside their manuscripts. Fact is, however, that authors can already do so, and a minority of the Protocols we publish do come with supplementary information in the form of videos. Clearly, we must do more to make our authors more aware of this possibility.

Going back to the posters, maybe unsurprisingly, Nature Protocols, in its yours truly incarnation, decided to award the prize for the poster describing the most interesting technique to Dr. Angel Mak, a post-doc in Puy-Yan Kwok’s University of California San Francisco laboratory, for the poster entitled “De novo genome assembly and structural variations detection by genome mapping in nanochannel arrays”. Of course, the technique described in the poster is none other than Genome Mapping in Nanochannels, already described in Dr. Kwok’s lecture two days earlier. My sincere congratulations on your research work Angel! But I had a wonderful time looking up all the posters and talking to several of the presenters. Very exciting work being presented by each poster, and thank you to all the researchers I spoke to, none of whom let transpire any disappointment at the dumbness of some of my questions.

It was Friday after 4 pm by the time I announced the winner of Nature Protocols’s prize, and it was time for me to catch a cab to go to the airport. I had to give a miss to the Irish Night — Whiskey Tasting Experience and Culture Night Performance Including Broadway Acclaimed Actor Eilin O Dea, organized for 5.30 pm that evening. A London night was beckoning for me; not that I had anything special planned, in fact I had nothing at all planned, but even with my relatively early flight from Belfast, I did not reach home till 11 pm that night. Sadly, I’m too old for anything more lively than that. And hey, now I’ve got an excuse to go back to Belfast and complete the Northern Irish experience…

Thanks a lot Mark and thanks a lot HGV2014 participants for the excellent time all around!

Nature Protocols authors receive the Nobel Prize in Physiology or Medicine

Congratulations to Nature Protocols authors James E. Rothman and Thomas C. Südhof, who alongside Randy W. Schekman have been awarded this year’s Nobel Prize in Physiology or Medicine “for their discoveries of machinery regulating vesicle traffic, a major transport system in our cells”, according to yesterday’s press release by The Nobel Assembly at Karolinska Institutet.

This year’s award acknowledges the fundamental contribution that research by Randy W. Schekman, James E. Rothman and Thomas C. Südhof has provided to the elucidation of the system that precisely controls the vesicle-based transport and delivery of cellular cargo within cells and from the cells to the outside environment. This traffic control is essential for the correct functioning of cells, and defects to it are known to occur, for instance, in diabetes and in a number of neurological and immunological disorders.

In detail, Schekman’s work has been instrumental in identifying the three classes of genes that control the cell’s transport system; Rothman, the author of two Nature Protocols articles, shed light on the mechanism by which the formation of specific protein complexes allows vesicles to dock and fuse with the correct target membranes; and, through his work of nerve cells, Südhof, the author of one Nature Protocols article, enabled the identification of the calcium ion–regulated molecular machinery that directs proteins to bind vesicles to the outer membrane of nerve cells, as a preliminary step to the release of neurotransmitters.

Dr. Südhof’s contribution to Nature Protocols is not specific to the research just mentioned, but it does provide detailed instructions on how to perform high-throughput gene expression profiling in individual neuronal cells using quantitative PCR. The approach covered by this protocol enables the investigation of hundreds of transcripts from a single neuronal cell, and it can be used to characterize, for instance, lineage-specific, reprogrammed neuronal cells.

One of the main areas of Dr. Rothman’s lab research is the protein superfamily known as SNAREs, which are present in yeast and mammalian cells. The key job of these proteins is to mediate fusion between vesicles and the cell membrane or the membrane of a cellular compartment. SNAREs are often subdivided into two categories, vesicle or v-SNAREs, which are embedded on the surface of vesicles, and target or t-SNARES, which are located on the surface of target membranes. Interaction between v-SNAREs and their t-SNARE counterparts directs the fusion of vesicles to target compartments through the formation of a three-protein complex formed by cognate SNAREs anchored on ‘opposing’ membranes.

The structure of a v-SNARE nanodisc

The structure of a v-SNARE nanodisc. The green ribbons represent copies of a v-SNARE.

Dr Rothman has co-authored two protocols published in our journal, both useful for the study of SNARE proteins. A 2012 paper describes the preparation of fluorescently labeled v-SNARE liposomes and t-SNARE–reconstituted planar, supported bilayers that can be used to monitor docking and fusion events by conventional far-field epifluorescence or total internal reflection fluorescence microscopy. One year later, almost to the day, we published a second protocol co-authored by Dr. Rothman. In this article the authors detail the preparation of nanodiscs of fixed (small) size that contain fluorescent lipids and copies of a v-SNARE. Upon fusion of one such nanodisc with a liposome containing the cognate t-SNAREs, the occurrence of a fusion event can be quantitatively monitored following the increase in fluorescence caused by the dispersal of the initially quenched fluorescent lipids from the nanodisc into the non-fluorescent, and much larger, interior of the liposome.

The fusion process between a v-SNARE nanodisc (bottom) and a t-SNARE liposome (top)

The fusion process between a v-SNARE nanodisc (bottom) and a t-SNARE liposome (top)

 

Well, our heartfelt congratulations to Randy Schekman, James Rothman and Thomas Südhof for their achievement. And please forgive my lack of modesty when I point out that it also feels good to have it confirmed yet again that Nature Protocols authors are among the very best in their fields.

My weekend at ESBOC

My three days at the European Symposium on Biological and Organic Chemistry (ESBOC) did not start in the most auspicious way. Or, again, maybe they did. Well, Friday the 17th, the day of my departure, is a day of bad luck in Italy, much like Friday 13th is in many other countries (it may not sound like it given my name, but I am indeed Italian), and my stopover at Birmingham New Street station to catch my connection to Newtown (Powys), Wales, certainly got my heart pumping.

To make a long story short, my apparently comfortable fifteen-minute leeway between my arrival from London Euston and my departure to (seemingly unknown in Birmingham) Newtown got eaten away by a mostly fruitless search for ‘my’ platform in a station, which, I later found out, is undergoing significant redevelopment — and is not very passenger-friendly. After a few minutes’ queue to reach the information desk, I got sent by the guy behind it to “platform 5b”, when I probably had something like seven minutes left to spare. After about five minutes of frantic soul-searching while pacing on a platform that just didn’t seem right, and that was at once platform 5, platform 5a and platform 5b, I grabbed my trusty trolley by the neck, ran up the stairs to the main concourse, then to the closest display panel, and finally down the stairs to platform 6b, as a train moved slowly past it. Was it my train? Was it pulling in? Was it pulling out? Well, suffice to say that, much to my relief, I made it to Newtown on time to catch the minibus meant to take all ESBOC participants arriving by train to the conference venue at Gregynog, a relatively isolated location about 8 km of narrow, windy road away from the town.

View of Gregynog. Courtesy of Dr. James Redman.

View of Gregynog. Courtesy of Dr. James Redman.

Gregynog… It is actually a beautiful country house surrounded by 750 acres of (beautiful, it behooves to stress) gardens and woods sitting right in the middle of Wales. It was donated to the University of Wales in 1960 by its owner, the late Miss Margaret Davies, as an arts center. As I started looking for my room, a friendly and slightly surprised member of staff exclaimed, “Room G2? It’s one of our two best rooms sir!” Wow, time to let my hair down, if I hadn’t just had it cut rather short, that is. The room was actually very nice and comfy, and I had something like half an hour to unpack before the conference reception and presentation.

This year’s annual conference was the 47th of the series, as this symposium was first organized here at Gregynog in 1967 by Professor Cedric Hassall of University College, Swansea. At the time, it was called Gregynog Natural Products Symposium, a name changed to European Symposium In Bio-Organic Chemistry in 1971, to become the present-day European Symposium on Biological and Organic Chemistry only in 2011. This year’s chair, Professor Kai Johnsson of EPFL Lausanne, Switzerland, could actually not be present, as his wife is expecting a baby imminently, so the role of hosts was entrusted to the capable hands of the Chairman of the Steering Committee, Professor Tim Bugg of the University of Warwick, and of the Treasurer and Secretary, Dr. James Redman of Cardiff University.

 

Cover of the program of the 1967 Gregynog Natural Products Symposium.Courtesy of Dr. James Redman.

Cover of the program of the 1967 Gregynog Natural Products Symposium.Courtesy of Dr. James Redman.

The title of this year’s symposium was “Chemical Probes for Cellular Processes”, a subject that elicited an excellent response of corporate and non-corporate sponsors and, according to Professor Bugg, the most international attendance of the symposium to date. This year’s ESBOC had 60 participants from eight different countries affiliated to both academic institutions and industry. The program included 18 lectures by invited speakers, ten of them plenary and two ‘named’, and it also included a 12-strong poster session. Among the scheduled speakers were Nature Protocols’s own authors Ben Cravatt of the Scripps Research Institute, Edward Tate of Imperial College London, and Carsten Schultz of EMBL Heidelberg. All the talks, which included lectures by ‘industry’ speakers Dr. Ivan Correa of New England Biolabs and Dr. Keith Wood of Promega, were up my alley, so to speak, as they were generally given by chemists who applied their skills to attempt to answer biological questions, exactly my research background.

Participants of the 1969 Gregynog Natural Products Symposium. Cedric Hassall is the second person on the left seated. Courtesy of Professor Timothy Bugg.

Participants of the 1969 Gregynog Natural Products Symposium. Cedric Hassall is the second person on the left seated. Courtesy of Professor Timothy Bugg.

It wasn’t just that I understood the talks in their entirety, which is undoubtedly a big plus, but I actually really enjoyed them. The symposium was small enough that no talks needed to be scheduled at the same time, so I was able to attend all of them. The lectures spanned subjects like applying bioluminescence to the study of intracellular interactions or the imaging of fatty acids, activity-based protein profiling, phage selection of peptides with different bio-relevant properties, the synthesis of riboswitches to control gene expression, the expansion of the SNAP-tag tool box, and fluorescence-based approaches to investigate nucleic acid structure. Talking about activity-based protein profiling, unfortunately Professor Ben Cravatt, who was supposed to give the first day’s Royal Society of Chemistry Bio-Organic Group Lecture entitled “Activity-based proteomics: applications for enzyme and inhibitor discovery”, could not be present, due to a family commitment, but he was replaced by post-doc Ken Hsu from his lab, who, despite the short notice and an 8-hour jetlag — Scripps is in San Diego, California — gave a wonderful talk on the use of activity-based protein profiling on serine hydrolases for the discovery of bioactive lipid networks by chemoproteomics and metabolomics.

The next day’s ‘marquee’ talk, the Cedric Hassall Lecture (“In vivo selective optical imaging of tiny tumors to rationally designed activatable fuorescence probes”), was to be given instead by Professor Yasutero Urano, of the University of Tokyo. Except that Yasutero was not the speaker’s first name. As he initiated his talk, Professor Urano pointed out that his name is actually YasuterU, and went on to speculate that the tendency of his European colleagues to spell it with an “O” must be due to an understandable mental association between himself and Elvis CostellO. Well, to be honest, I did not see much of a physical resemblance between Professor Urano and Elvis Costello, not least because the former is obviously much younger. The association was made clearer, however, by the thunderous applause that greeted the end of Urano’s lecture. A really impressive talk, even for an outsider like myself.

Professor Urano had described his laboratory’s work on synthesizing and testing probes that become fluorescent only when taken up by tumor cells, a strategy that would prospectively enhance surgeons’ ability to literally see cancerous tissues when operating. Urano’s research group had used two main approaches to achieve their goal. In one, pH-sensitive fluorescent probes are fused to an antibody specific to membrane receptors that are highly expressed on the surface of tumor cells. Upon endocytosis, the low-pH environment of the cell lysosome causes activation of the probe, which thus becomes fluorescent. In the second, the probe is fluorescently inactive as long as it remains bound to an appended peptide group, which can however be cleaved by GGT, a peptidase highly expressed on the surface of tumor cells. Upon cleavage of the peptide bond, the newly fluorescent and highly hydrophobic probe concentrates first on the cell membrane, and, eventually, inside the cell, to highlight that cell’s unhealthy state. What did I say? Really impressive.

As ESBOC drew towards its end, I even found out how Gregynog is pronounced correctly. Two competing schools of thought seemed to take shape on the matter at the conference. According to participants with Welsh connections, the pronunciation in southern Wales would be something like Greg-EYE-nog, whereas people in northern Wales may actually pronounce it Greg-E-nog. Unfortunately Gregynog is almost in the dead center of Wales… Mystery solved on Saturday night, however, when we ESBOC participants were entertained by the charming music of Welsh harp player and folk singer Siân James, clearly a reliable source on all things Welsh. To my untrained ear, she pronounced the locale “Greg-e-nog”, making a slight clicking sound with her tongue when she uttered the second and third g’s of the word.

What can I say? My experience at ESBOC was excellent all around. The place was fantastic, the talks outstanding, the participants friendly and chatty, and my ‘contacts’, Professor Bugg and Dr. Redman, always helpful. Even the food was terrific. Word on the street was that Gregynog had recently hired a new German chef, and everyone was raving about him. Ok, I am Italian, but I have to admit that he really knew his stuff. On Sunday the weather too smiled on us, with a gloriously sunny day; too bad that we were about to leave and that strolling around the grounds had been scheduled for the day before. Oh well, not bad for a conference two train and one minibus ride away that started on a Friday the seventeenth…

Well, the last bit of important information concerns next year’s ESBOC at Gregynog: ESBOC 2014 will take place on 16–18 May, and it will be organized by next year’s scientific chair, Dr. José Luis Mascareñas of the Universidade de Santiago de Compostela, in Spain. The title of that conference will be “Biological Applications of Supramolecular Chemistry”. Make time to go, if you can.

 

 

 

At MASAMB XXIII

So I’m back… Well, I have actually been back for a little while already, as this year’s Mathematical and Statistical Aspects in Molecular Biology (MASAMB) meeting was held last April 11th and 12th.

Mission accomplished, it is fair to say. The idea was for me to acquaint myself face-to-face with research areas — bioinformatics and statistical genetics — which Nature Protocols has always covered, and in which it hopes to be even more active in the near future. For sure, over a day and a half I had the opportunity to attend a total of 25 presentations divided into five sessions: statistical bioinformatics, computational cell biology, next-generation sequencing, systems biology, and evolution. I will not claim to have understood everything I heard, but all presentations were stimulating and thought-provoking. Given my background in ‘wet’ chemistry and biochemistry, I guess it was no surprise that I found it easier to follow the biological side of the talks than the parts that dealt with mathematical and computational models, but I am now much more familiar with the landscape in this research area.

My attendance of MASAMB, as I discovered on site, was primarily made possible by the engaging and amiable host of this year’s MASAMB, Dr. Michael Stumpf of Imperial College, via the intercession of my email contact, Dr. Louise English. During one of the poster sessions, Michael explained to me that one of the main goals of MASAMB is to provide PhD students and post-docs with a workshop-like venue in which to present their own research in an environment that is more ‘intimate’ and less intimidating than the average meeting. Students and post-docs are those most encouraged to present their work, Michael added, and most of the conference speakers belong to this category.

Besides the scientific quality of the presentations, I was quite impressed with how well they were timed and organized. Each talk was to last 20 minutes, including eventual questions from the audience, and not a single presentation out of 25 went long, while the switch between speakers always ran smoothly and quickly. I found this to be quite the impressive feat, both in terms of the technical prowess of the meeting organizers and of the scholarliness of the young — and one would assume generally relatively inexperienced — speakers.

This year’s MASAMB conference attendance was about 110, toward the high-end of the average, but it still felt quite ‘cozy’ for participants and reporters alike (well, for the only reporter, me). Speakers were mostly from UK institutions (e.g., Imperial College, UCL, University of Glasgow, University of Warwick, University of Bristol, Cancer Research UK, Brunel University, University of Manchester, University of Sheffield and the University of Reading), but there was also a significant representation of researchers from other European countries, including Cyprus (University of Cyprus), Germany (EMBL Heidelberg), Austria (Center for Integrative Bioinformatics and the University of Veterinary Medicine), Switzerland (ETH-Zürich), Poland (Polish Academy of Sciences), and Finland (Aalto University).

If I were a young scientist being trained in computational biology or bioinformatics in a European institution, I would try my best to attend MASAMB, and apply to either give an oral presentation or present a poster on my own research. Talking about posters, the poster count for this year’s conference was 35, which, according to Michael Stumpf’s concluding remarks, included “not a single stinker!” The high quality of the pool made it all the more impressive for the winners of this year’s best poster competition: Ann Babtie, Dominic Smith and Phoebe Jones from Imperial College for “A moment expansion approach for stochastic simulation and inference” and Agnes Jonas from the University of Veterinary Medicine of Vienna for “Modelling allele frequency trajectories of experimental evolution with fruit flies”. On presenting the awards, Stumpf remarked how pleasantly impressed he was that three of the four recipients were women, even though the vast majority of entrants were men. Well, you heard it, if you’re not just a young scientist, but also a woman, it’s time to do your best to attend the upcoming MASAMBs and extend the flattering gender-slanted tradition.

Well, attending MASAMB XXIII was an excellent experience, which would not have been nearly as pleasant or useful if it weren’t for the current and former members of the Stumpf laboratory. Aside from Michael himself, I had the opportunity to meet and converse (about science and everything else) with research fellows Tina Toni and Sarah Filippi, post-doc Juliane Liepe, graduate students Ann Babtie, Adam McLean and Delphine Rolando, and former Stumpf group member (now a post-doc at the University of Cyprus) Kamil Erguler. I sincerely thank them for their invaluable help and for making me feel very welcome. I feel Nature Protocols and I have now an ‘in’ into the world of bioinformatics research, and I look forward to receiving news and feedback on how Nature Protocols can be of more help to researchers in this field — right Juliane?

One last bit of information: the general preference is for the yearly MASAMB meeting to take place in the UK for two consecutive years, then somewhere else in Europe, before returning to these shores. Tentatively, MASAMB 2014 will take place at the University of Sheffield, a venue that was preferred by this year’s participants to the other two candidates, Warsaw and UCL. Stay tuned…

Nature Protocols at the 23rd MASAMB annual workshop

With the advent of technologies that have made large-scale genome sequencing projects possible and since a stream of high-throughput biological assays have been developed to collect, for instance, data on proteomics, metabolomics and gene expression, the need for ever more advanced, powerful and sophisticated bioinformatics tools has become an increasingly stringent fixture of today’s molecular biology research.

The Mathematical and Statistical Aspects in Molecular Biology (MASAMB) meeting has brought together every year since 1989 a relatively small number of researchers (usually around 100) from the fields of mathematics, statistics, computer science, bioinformatics, and biology to discuss and exchange ideas on methodological and applied research in an “intimate” workshop setting. This year’s MASAMB will take place on April 11th and 12th at Imperial College London, with the main topics of discussion being statistical bioinformatics, computational cell biology, next-generation sequencing, systems biology, and evolution.

Since their birth in 2006, both Nature Protocols and the Protocol Exchange (which was originally called the Protocols Network) have published a steady flow of protocols that can be categorized under the general term “computational biology”. Unsurprisingly, some of these protocols have proven very popular with our readership, with a number of these articles ranking among the journals’ most cited papers.

As we anticipate the number of published protocols in this field to increase further — as well as the interest in them from our readers — for the first time this year, we have decided to attend as a journal MASAMB, sending along one of our associate editors, well, sending along myself, Baldo Lucchese. I will be covering the workshop for Nature Protocols attending as many talks as possible and speaking to as many researchers and, hopefully, readers as humanly feasible. The objective will of course be to gain the wherewithal to make Nature Protocols a more helpful source for researchers in bioinformatics and statistical genetics and for readers with an interest in these fields. It will also be an excellent opportunity for me to meet in person a few of our authors. I certainly look forward to that too.

If you too are going to be at the conference, please do come by to say hi.

Stay tuned for the next blog post on this issue after I come back from MASAMB XXIII.

Following science’s lead to reflect on the ethics of mitochondrial transfer

When I first mentioned to some colleagues that I was thinking of writing this post in the journal blog, a few quizzical expressions surfaced on the faces of Nature Protocols’ editors. After all, the ethical and philosophical implications of the protocols we publish aren’t the usual remit of Nature Protocols. Yet, when I found out that a method for mitochondrial DNA ‘transplantation’ introduced by Oregon Health & Science University’s Shoukhrat Mitalipov and co-authors is now technically almost ready for the fertility clinic1, my mind started to wander into all kinds of questions and considerations that I thought I’d like to share with my colleagues and the readers of this blog.

In 2010, Nature Protocols published “Chromosome transfer in mature oocytes2, by Tachibana, Sparman and Mitalipov, an article that details the transfer of chromosomes from the mature oocyte of a rhesus monkey to the enucleated egg from a different rhesus monkey, so that the resulting oocyte has the nuclear DNA of a female primate and the mitochondrial DNA of another. This protocol was based on the ground-breaking 2009 Nature paper “Mitochondrial gene replacement in primate offspring and embryonic stem cells3, which also reported that healthy monkeys had been born as a result of the procedure. These were monkeys that displayed no detectable presence of mitochondrial DNA of their biological mothers, but only that of the oocyte cytoplasm donor.

Mitochondria are often — and possibly too simplistically — called the cell’s power plants, because it’s in these cytoplasmic organelles that most of the ATP, the organism’s energy currency, is produced. Mitochondria have their own DNA (two to ten copies per organelle), which, as opposed to nuclear DNA, is passed en bloc from mother to offspring, without any paternal contribution.

Because of their role in ATP synthesis, mitochondria are exposed to a high concentration of free oxygen radicals, which, in conjunction with a lack of histones and limited mitochondrial DNA repair mechanisms, possibly explains why mitochondrial DNA mutations occur at a tenfold-plus rate compared with nuclear DNA mutations3. In humans, serious and often fatal disorders caused by mitochondrial DNA mutations affect 1 in ~4,000 children. Although current treatments alleviate symptoms and slow disease progression, no cures are available for these mitochondrial diseases.

At least in principle, a method that enables the complete replacement of mitochondria in the egg or embryo from a woman with known mitochondrial DNA defects with mitochondria from a donor with no such defects could act as an effective, ‘pre-emptive’ treatment of diseases linked to mitochondrial dysfunction. Just recently, Nature published another paper by Mitalipov et al.1, which reports how the approach described in the Nature Protocols 2010 paper2 has been successfully implemented to produce normally fertilized human zygotes that contained mitochondrial DNA only from the donors of oocyte cytoplasm (and not from the nuclei donors). These zygotes were found capable of developing blastocysts and of producing embryonic stem cells, which suggests that, if implanted in the womb, they could develop into healthy babies.

Of course, a breakthrough of this magnitude gives hope that a clinical application may not be too far down the road. This optimism is further encouraged by the significant success in achieving similar results in terms of mitochondria replacement in human embryos4 — albeit via a different approach — by a research group based in Newcastle, UK, and led by Professor Doug Turnbull. Incidentally, Dr Turnbull and co-authors published a method for the transfer of nuclear genome as a promising approach for the prevention of transmission of human mitochondrial DNA disease5 in 2010 in the Protocol Exchange.

The optimism, however, is tempered by substantial legal obstacles to the clinical application of these approaches. For instance, UK law currently forbids the genetic modification of human embryos or human eggs for treatment purposes, which prevents clinical use of both the approach developed by the UK-based group and the approach developed by the US-based one. In the US, the NIH restricts funding for research that destroys human embryos, so Mitalipov’s research group had to conduct its research using money from private donors6.

These legal and procedural hurdles are not mere technicalities, and a number of fundamental ethical and philosophical questions have to be univocally answered before the medical community embarks on the clinical use of pre-natal mitochondrial DNA replacement. Mitochondrial DNA only encodes 37 genes, or about 0.2% of our entire genetic make up. But the question still stands: what will be the exact relationship between a child and the woman to whom that child’s mitochondrial DNA originally ‘belonged’? Surely children who have not received their mitochondrial DNA from their biological mothers will look like their parents, but arguably efficient mitochondrial activity is vastly more important for the biology of a human being than the color of his or her eyes or whether his or her hair is straight or curly.

Furthermore, replacing a mother’s mitochondrial DNA does not make a difference just to her children, but given that it is passed down, more or less intact, from generation to generation along a matrilineal route, mitochondrial replacement may have permanent effects on many generations to come, including any possible unforeseen adverse consequences of the procedure. In order to observe, probe and record any such potential long-term adverse effects, subsequent generations of people who owe their mitochondrial genetic makeup to mitochondrial replacement will most likely have to be enrolled, basically at birth, in long-term, follow-up medical studies for decades to come; but presumably, once the age of consent has been reached, these individuals will have the right to refuse participation in such studies, won’t they? And what about their personal lives? Given the medical implications and the health concerns for their offspring, are the people involved going to be expected to disclose their genetic origin to partners they may want to have children with? A case could easily be made that the latter question applies only to female descendents of the woman who underwent the initial mitochondrial replacement. And shouldn’t the inherently discriminatory nature of this uneven burden be an additional cause for profound and unsettling moral questioning? After all, men are essentially genetic dead-ends when it comes to their mitochondrial DNA.

These are questions that no sensible person would ever volunteer to answer unless they had to, but we find ourselves exactly at that point in time. An answer must be sought and given, as the suffering of many men and women, children and babies may be avoided and their early death averted by the implementation of procedures to replace mitochondrial DNA. The UK government has launched a national, public consultation on mitochondrial replacement, which is to run until Friday December 7th 2012, and which will advise whether change in legislation is appropriate. At the very least, this consultation must serve to stress the fact that in democratic societies, a decision on whether to go ahead, and in which terms, with the clinical application of these techniques, ultimately rests with the will of its members.

I argue that the scientific community at large should feel a particular responsibility to contribute to the moral, ethical, and philosophical discussion that is taking place. Scientists, researchers and science experts in general are among the people who are best equipped to know, understand, and anticipate the wide range of implications and ramifications of applying a technique such as mitochondrial replacement to the treatment of mitochondrial diseases. Ultimately, the Nature Protocols blog is as good an informal venue as any to discuss these matters and to air one’s views on the trove of thorny questions forced open by the publication, among other articles, of protocols in Nature Protocols2 and the Protocol Exchange5

 

 

1.                  Tachibana, M. et al. Towards germline gene therapy of inherited mitochondrial diseases. Nature https://dx.doi.org/10.1038/nature11647 (2012).

2.                  Tachibana, M., Sparman, M. & Mitalipov, S. Chromosome transfer in mature oocytes. Nature Protocols 5, 1138–1147 (2010).

3.                  Tachibana, M. et al. Mitochondrial gene replacement in primate offspring and embryonic stem cells. Nature 461, 367–372 (2009).

4.                  Craven, L. et al. Pronuclear transfer in human embryos to prevent transmission of mitochondrial DNA disease. Nature 465, 82–85 (2010).

5.                  Craven, L., Tuppen, H., Taylor, R., Herbert, M. & Turnbull, D. Pronuclear transfer in abnormal human embryos. Protocol Exchange https://dx.doi.org/10.1038/nprot.2010.54 (2010).

6.                  Cyranoski, D. DNA-swap technology almost ready for fertility clinic. Nature https://dx.doi.org/10.1038/nature.2012.11651 (2012).