Phone talk with Raymond L. White

    (Howard Cann et al., 16 sept. 2010 at Génopole d'Evry, proof read by R L White, jan. 2012)


    DR

    See also : S. M. Prescott, J. M. Lalouel, M. Leppert, 'From Linkage Maps to Quantitative Trait Loci: The History and Science of the Utah Genetic Reference Project' (Ann. Rev. Genom. Human Genet. 2008.9.347-358) and D. Botstein, R. L. White, M. Skolnick, R. W. Davies, 'Construction of et Genetic Linkage Map in Using Restriction Fragment Length Polymorphisms' (Am J Hum Genet, 32.314-331, 1980).


    H. C. : This program - the idea of Jim Watson and people at Cold Spring Harbour I was told - has resulted in a number of countries participating in trying to get the history down and known about the Human Genome Project. There was a committee set up in each of these countries and certain in France. The Chairman of the committee in France is Jean Weissenbach. I am a member of this committee. I think we would like to start pretty much at the beginning, in the early 1980s. Ray, you were, as I understand it, very interested in doing gene mapping with Restriction Fragment Length Polymorphisms (RFLP). This was at the heart of a paper by David Botstein, yourself, Ron Davis, and Mark Skolnick.  CEPH had just gotten funds, Jean Dausset had gotten funds and there was also a decision to get interested in gene mapping, the genome, in fact. One important question that I have for you relates to your first meeting with Jean Dausset. What happened at your first meeting with him and Daniel Cohen? This, I imagine, was well before that famous meeting in 1984, in Paris, in October.

    R. W. : There were two meetings, three if you include the phone call. We were setting up a Howard Hughes Medical Institute (HHMI) small meeting in Miami at the HHMI headquarters to talk about the new genetics. Anyway, prior to the meeting I got a phone call from a guy with a strong French accent, who described himself as Jean Dausset, Nobel Prize winner.  He wanted to be invited to the meeting. That sounded fine to me.  I could not quite get what he was talking about, but it still seemed like a fine idea. He came to that meeting and made a presentation about unity, human leukocyte antigen (HLA) and such. That was all fine.  He did not technically meet with me until a meeting that occurred a little after that, at a subsequent larger, more general meeting in Miami. At that meeting, I met Jean-Marc Lalouel and Daniel Cohen. After I made my presentation, Lalouel came up and said, ‘We must talk.’ He then went on to explain to me that he had actually solved all of the problems of analysis of human genetic data with his new programme. This was in a taxi cab as we rode around Miami. That sounded really interesting to me and proved to be enormously helpful, because we were then able to recruit Lalouel to Utah, where he set up the analysis section of our mapping efforts. All of that was direct or indirect.  There was another meeting in Paris after that.

    H. C. : There was a meeting in Paris in 1984, October.

    R. W. : I do not know if there was a meeting, exactly, but I did visit Paris. It was in Jean Dausset’s offices. I was talking to people. Jean Dausset was there, as was J. M. Lalouel and D. Cohen. Regarding the 1984 one that you are talking about, remind me of exactly what that was.

    H. C. : That was when you came to Paris, but there were others with you. David Botstein was there, Helen Donis-Keller, Ken Kidd, Dick Getty were there.

    R. W. : Those were all after those first meetings.

    H. C. : That was after. Apparently Ray, from what I understand, somehow you were involved. This was when Lalouel was in Paris and working with Daniel Cohen and Dausset to get CEPH into the act.

    R. W. : Right.

    H. C. : They realised that they had families and at the Miami meeting, they learned about your families, the Utah families. Somehow, you, Dausset, Cohen and Lalouel got together and decided to put the families together, into a project. This is what I understand. Does that sound familiar to you at all ?

    R. W. : It does indeed. I willingly agreed that what they were setting up would offer a good way to distribute the genetic family resources that we had accumulated in Utah.

    H. C. : You thought their work was a good idea. Did you ever question it before ?

    R. W. : No. I never questioned it before, nor after, but a Utah colleague made the public assertion that it was one of  the biggest public-relations ‘faux pas’ in scientific history.  However, I pointed out that we had limited resources that we could apply to distributing families, which Dausset wanted to do, or we could use those resources to try to find cancer genes. I chose to look for cancer genes, which I still believe was the right choice for my research group in Utah.

    H. C. : However, that was later.

    R. W. : But, that was already clearly a major goal of our mapping effort.

    H. C. : Did you have the impression that Dausset was the brain behind this. Let me put this bluntly : was Cohen the brain behind this, or Lalouel, from the French side?

    R. W. - Well, I would say that the germination came from Dausset. The plan was very analogous to what he had done with HLA, to encourage investigators to form a collaborative group, sharing the basic materials. That was likely. I do not know if he ever said that, but it was transparent that that was the roadmap for what he had in mind.

    H. C. : However, do you think the application of that to linkage mapping related to Dausset ?  Do you think that was Jean-Marc ?  Do you think that was Daniel ?  Do you know ?

    R. W. : None of the above.  The linkage mapping was the reason we collected the families in the first place; we wanted to make a map of the genetic markers. We had that clearly in mind at the beginning. That is one of the reasons why I think Dausset’s plan fitted in very well with us.  We were interested in seeing a global map constructed. I do not believe that Dausset had that in mind at that early point – his perspective seemed more abstract. I do not know what he had in mind with these families. However, that pretty quickly became the motivation for collection of the CEPH families.

    H. C. : Okay, but I do not think I closed the question well. From the French side, where did the interest in linkage mapping with those families come ? Did it come from you ?

    R. W. : When I arrived at the Miami meeting, where I first met Lalouel and Cohen, I already knew linkage mapping was the purpose for the families. I thought the French families had perhaps been collected for some other purpose, but I don’t remember discussing that specifically.

    H. C. : OK. Let us go on to the meeting with Dausset in his office in Paris. Were Cohen and Lalouel with him ?

    R. W. Lalouel, on the other hand, was very sharp, very quick and impressed me a great deal. Cohen was also there. I remember that in particular, because he volunteered a lesson on heteroduplex heterozygotes and how that could affect recombination. Since that was what I had done my PhD thesis on, I was able to follow the lecture easily.   

    H. C. Was that after the Miami meeting or before ?

    R. W. : Miami was the meeting where I first met Cohen and Lalouel. This was after.

    H. C. : It was my impression that it was a pretty enthusiastic meeting. People were very interested; they participated well and there was some solidarity. I do not know how to say that.

    R. W. : Absolutely. It was a bunch of people who were going to get a great Christmas analyzing the family collection.

    H. C. : Naturally, they all signed up. Dausset had a great sense of genius, to create an international collaboration out of what was in fact a key party collaboration.

    R. W. : I agree. I think he was very sharp in doing that.

    H. C. : That is why we are talking to you. I should tell you, we are going to contact Jean-Marc too. From the French side, was it Dausset ? Was it Dausset and Cohen, or Dausset, Cohen and Lalouel ? Did Dausset realise the importance of families for linkage, or was that Cohen, or was that Lalouel ?  Can you tell us?

    R. W. : I think, regarding families for linkage, that the first recollection I have was in the taxi ride with Lalouel and Cohen. Lalouel must already have been thinking about families and genetic linkage for the future or he would not have developed his computer programme for linkage analysis.  Whether he has intending to link markers to diseases directly or start first with a map of the markers is not clear to me.

    H. C. : Did you start doing work on human linkage maps before you came to Utah ? I thought you came to Utah because of it.

    R. W. : The mapping with three-generation families came to me following a small Cold Spring Harbour meeting. It had occurred to me that just as with any other mapping expedition, if you selected families with the right characteristics, it would make mapping of the markers much easier. Three generation families, consisting of grandparents, parents and as many siblings as possible. That was a clear, distinct idea by the time I got back to Utah from the Cold Spring Harbour meeting, and I set about, with Mark Leppert in Utah, to begin ascertaining those families.. The work before Utah was that with with Botstein, developing the idea of creating a large set of Restriction Length Polymorphism (RFLP) markers. Such markers could be used to map a disease. However, the general idea of creating a map of the markers is in the original Botstein, White, Skolnick, and Davis paper in the American Journal of Human Genetics and the idea of making a map of the markers through linkage was intrinsic to our interest in developing the markers described in the paper

    H. C. : Do you have any idea of the date of that ?

    R. W. : The paper was published in May of 1980, and was titled “Construction of a Genetic Linkage Map in Man Using Restriction Fragment Length Polymorphisms”.

    H. C. : After the October (1984) meeting (in Paris), things got started. I know we got cell lines from you and from a lady named Lesley, remember her?

    R. W. : Lesley Jerominski, yes. She was a master tech, who did the work of creating the cell lines.

    H. C. : CEPH got started making DNA and then distributing it. Things got started. Lalouel set up the database. He distributed the linkage programme with Mark Lathrop, who you know well. I am sure.

    R. W. : Yes, indeed.

    H. C. : I always had the impression that you were part of the real backbone of that project. You were very supportive of the project; you certainly participated to the hilt in that project.

    R. W. : That is my recollection too.

    H. C. :  Up to that time, was there a point early on when you started to ask questions about that collaboration? Was it really the thing for you to do ? Was it really the best approach ? This was the collaboration of Utah families, French families and Venezuelan families.

    R. W. : I do not remember having any doubts about it. Howard, that is a strange question. Why do you ask that ?

    H. C. : I am thinking about the Donis-Keller paper.  You were pretty angry about it, I can tell you. I remember you got me on the phone and wrung me out.

    R. W. : She used our data inappropriately, in my view, to make her map. She was a difficult person.

    H. C. : That is a good way to put it. Afterwards, after that paper appeared, did you still feel that the CEPH collaboration was the way to go ?

    R. W. : Yes. Just because one person or group abuses it, is not a good reason to shut it down.

    H. C. : Then the other thing we are interested in is that NIH finally got very interested in the linkage with CEPH, the collaboration in the linkage map. Do you remember that ?  It was about the time Watson became the Tsar of mapping at NIH.

    R. W. : There was something about that. I remember he was pissed off at me because I was not doing things directly or quickly enough. I know (Watson) questioned me closely on whether we should be doing chromosome-by-chromosome maps, or whether we should try to do the whole thing globally. I was still in the mode of being concerned that we would have enough markers etc. I was doing chromosome-specific maps from cell hybrids, from DNA from cell hybrids. I told him that I thought chromosome by chromosome was the way to go, but in retrospect, I think that was a wrong decision on my part. It would have been better to just keep making lots of markers and let the chromosome match emerge from the linkage data itself.

    J. F. Prudhomme : After the CEPH meeting, how did you decide the budget ? Did you decide that each country had to pay for the collection of their own families ? What happened ? That would seem a reasonable plan, that if somebody wanted to submit families, they would have to collect them themselves. I am not sure if the families had already been collected by that point and it was more of a matter of transferring them from Utah to Paris.

    H. C. : I will try to answer this question. I think that each participant in the collaboration had their own budget and that budget came from various sources. CEPH had the budget from Madame Anavi.  Ray can tell you that Utah is better than anywhere else. I imagine you were NIH-funded, Ray, but I do not know. Did you have the feeling that NIH supported the idea of the collaboration pretty well ? We have the idea that NIH felt that maybe they should take over the map, which may be because we are in France. Do you remember anything like that ?

    R. W. : I do not think I paid too much attention to that. As I recall, our NIH support was primarily for marker development.  Maybe you could include mapping in the proposal. We did get a good chunk of NIH money for marker development.   I believe we supported the collection of the families and creation of the cell lines with HHMI support.

    J. F. P. : At this Paris meeting, you talked about families, who had probes (sondes) and markers.

    R. W. : Yes. There are two parts to this. One is families and the other is identifying genetic variance to use as markers.  There was more participation in the marker identification. That is certainly what the major part of the large collaboration was, not it was family collection. The great majority of families were collected on Utah and there were some French families.  Most of the CEPH participants, however, were involved in marker development and the mapping of those markers. That was the beauty of the collaboration; they could map them using the CEPH resource, but they were supposed to report the data back so that other people could use them as well.

    J. F. P. : You sent your results to Jean-Marc Lalouel.

    R. W. : We sent our family DNAs to the CEPH in Paris. Jean-Marc Lalouel had been recruited at some point to come to Utah. The results he worked on were primarily developed when he was in Utah,although he may have started before he left Paris.

    H. C. : I think that is correct. Jean-Marc set up the database and the rules for using them and it is conceivable that in early 1984 and 1985, he may have touched some of the data that was coming in.  I think he was well-established in Utah when the database really took off…

    J. F. P. : Who developed the batch program ?

    R. W. : It was Jean-Marc and Mark Lathrop together. My impression was that Jean-Marc was the senior person and Lathrop was his junior fellow. Lathrop certainly did a lot of the pick-and-shovel work and Jean-Marc had a major part in the overall leadership. Both were highly expert. Jean-Marc Lalouel,   was trained in deep statistics, genetic linkage and that sort of thing.  He spent 10 years in Hawaii with Newton Morton

    H. C. : The other thing is that anyone in the collaboration who wanted to analyse the data could analyse the data. That was the understanding. There was not only Lathrop and Jean-Marc, but also a guy called Phil Green, who worked with Helen Donis-Keller. He did the analysis.  Now, other people did analysis too.  The idea was to put the data in a database and then make that available to the members of the collaboration…

    J F P. : It was really to share both families and markers.

    R. W. : That is right.

    J F P. : After 1984, You decided about meetings; you decided that you had to meet each year. What happened regarding that?

    R. W. : Yes, we agreed to have future meetings.

    H. C. : We did have future meetings. You may remember that you were on our advisory committee for the collaboration. It was the advisory committee, together with Dausset, Cohen and myself, who decided when to have the meetings. We had several meetings.

    R. W. : Yes.

    H. C. : They were not always in Paris.  At the time of the human gene-mapping organisation, we also had meetings. The group grew from 13 collaborators to a final figure of over 100 in the 1990s. In fact, we never set a limit on it; we never set a date, but little by little, as the linkage map took form and became known, it was used to make the physical map. There was less interest in using the families; I would think there was more interest in using the markers that were linked to detect disease genes.

    R. W. : That is right.

    H. C.  : You detected a very important one, I remember.  It was Neurofibromatosis One (NF-1), was it not ?

    R. W. : Publication of the mapping of the NF-1 gene was published almost at the same time by Seizinger et al in the June 5 Science and by Barker et al in the May 29 Science.

    H. C. : However, you were involved in mapping a number of disease genes, I think, even in mapping them.

    R. W. : Our first gene mapped was a gene for a form of colon cancer, familial polyposis  coli.  That came out on December 4, 1987.

    H. C. : We started out in the collaboration with RFLPs and then all of a sudden, micro-satellites were invented. There was a very important input in the use of micro-satellites for linkage mapping from your lab. Regarding the micro-satellite story, Jim Weber became a collaborator, because he wanted to use the families with micro-satellites that he had developed.  I was under the impression that you were pretty much on the ground floor at your lab in Utah. This was also with Variable Number Tandem Repeats (VNTR), micro-satellites or whatever you want to call them. Do you want to tell us about that ?

    R. W. : I think the VNTRs probably preceded the micro-satellites, but the micro-satellites were seen as a technical advantage, because you could score them by PCR.

    H. C. :  What is your distinction between a VNTR and a micro-satellite?

    R. W. : A micro-satellite has got a repeat length of three or four bases. A VNTR can have a repeat length of eight, 10, 15 pb.  We liked the VNTRs because they also provided multiple alleles and were easier to score than the microsatellites.  However, the ability to score the microsattelites by PCR was the technology that won out in the end.

    H. C. : Do you remember that story about the Venezuelan families of Nancy Wexler ? It came to a head at a Human Genome Meeting (HGM) meeting. Many of the CEPH collaborators were there and you were there. Nancy Wexler made a presentation with the idea of pooling the CEPH Utah families with her Venezuelan families. She proposed to put to a vote of the collaborators who were there and the vote was overwhelmingly against adding the Venezuelan families to CEPH. There were a lot of markers on the CEPH families already and it would just be extra work to put the markers on Venezuelan families. That is what I thought. Did you think that was a good idea ?

    R. W. :  I do not remember that as very significant one way or the other. That was kind of a technical decision.

    H. C. : Then next about CEPH, how did you and Daniel Cohen get along ?

    R. W. : I do not think we had any real issues.

    H. C. : This was throughout the collaboration?

    R. W. : Yes.

    H. C. : The collaboration kept going until maybe 1994. I know that you started using VNTRs in your map, as did Jean Weissenbach here in Paris. After the Weissenbach maps got made, there were three papers on that, with increasing numbers of micro-satellites. Then I remember a collaborative paper with a lot of CEPH collaborators, such as Ken Buetow and Jeff Murray. They were all about the CEPH families; the Weissenbach work was on CEPH families, but it was only on the eight biggest CEPH families. Do you remember, we had this agreement that people who used the CEPH families would do them all ?

    R. W. : Yes, I do remember it.

    H. C. : I cannot remember, but I wonder did that decision of Weissenbach and Cohen to just use the eight biggest families for the micro-satellite maps bother you?

    R. W. : Given the reasons that we had originally made it a requirement that you do all the families, I think I was slightly annoyed, but I did understand why they were doing it that way. They wanted to get a quicker map with a lot more markers and with lower resolution. We are willing to accept the lower resolution. Again, I do not remember being terribly concerned one way or another.

    H. C. :  You did not come to blows with Daniel Cohen about that.

    R. W. : Not that I can recall. I do not think I was at odds with anybody except perhaps Helen Donis-Keller.  But it does drive home the point that there was never a decision to punish anyone who ignored the rules by, for example, tossing them out of the collaboration, or even pointing a finger at them.

    H. C. : Ray, is there anything else you want to tell us ?

    R. W. : I just remember it as a time of very intense and focused effort to do a specific thing, which was to make a map of genetic markers using linkage. This was in order to identify the locations of human-disease genes. It was really the human-disease genes that were the drivers behind all of this.  We were making tools that would enable that.

    H. C. : You are right on that. I think I probably over-emphasised the mapping aspect. There was no question that many people in the collaboration felt very strongly about the disease-gene aspect and I think it was very successful from that viewpoint. A lot of papers came out. But let me ask you, looking back on this, was it the right thing to do ?

    R. W. : It was absolutely the right thing to do. It filled a very important gap. Many people think it was the beginning of implementation of the Human Genome Project.

    H. C. : Do you think it was ?

    R. W. : I do, yes. The principal feature was that it was worth investing in a large-scale project in order to create the tools you need to get at this very complex data, the human genome. The markers and the maps were the start of it. Now we are on complete sequences. I think it is just outstanding; it was very useful to initiate it and I am pleased to have been a major part of the effort.