When did you first know that you wanted to be a scientist?
I grew up in Australia and you had to choose your area of interest as history/geography/English, or sciences at age 14, and I picked the sciences. As I progressed through high school I thought that the biological and biochemical sciences were what I was most interested in, in a vague high-schoolish way.
What brought you from the University of Melbourne Australia to the then Department of Medical Genetics at the University of Toronto for your postdoctoral work?
My husband Evan Richard Stanley, who was a postdoc in Australia and stayed there a little longer than he might have if he was not waiting for me, went on a world tour, which is what Australian scientists did in those days, to give seminars all over the place. He visited Toronto because he was very interested in colony stimulating factors and James Till and Ernest McCullough were pioneering scientists in that area working at the Ontario Cancer Institute (OCI). There were other possibilities like San Francisco and also Boston, but when we looked at what might be possible for me in those places, I thought that Lou Siminovitch’s new area of somatic cell genetics sounded exciting. And Lou had also been involved in the hematopoietic stem cell work at the OCI, so there was a connection there. So I applied to him.
What was the most important discovery from your postdoctoral work with Lou Siminovitch?
Finding out the molecular basis of the first mutant that I isolated, which was eventually called Lec1. In the beginning it was called L-PHA resistant mutant, but the cells were also resistant to several other lectins, so it got a little complicated. After I left Toronto, I wrote a review where I called all the mutants by a simple Lec name. I worked on that first mutant a little bit with Rudolph Juliano who was at Sick Kids I think at that point, and found out that on SDS gels the mutant protein profile was all shifted. Together with the fact that the cells were cross-resistant to a lot of different lectins, even though I selected them with only L-PHA, and were hypersensitive to yet another lectin, suggested changes that were affecting many proteins, and this really suggested a glycosylation change.
In discussing this with Lou, he realized his swimming companion Harry Schachter, would be a perfect biochemist collaborator. So I collaborated with Harry’s lab at Sick Kids. I went over there, learned their assays and worked with his postdoc Saroja Narasimhan. We published our discoveries in PNAS showing that the enzyme activity missing in Lec1 was a particular N-acetylglucosaminyltransferase, which became GlcNAc-T1. That was the exciting bit. At that point no cloning was being done, as it was 1975. Eventually, when I cloned Mgat1, the name for the gene encoding GlcNAc-T1, we worked out the mutations in a series of Lec1 lines that I had isolated, and found that they all had a mutation in the Mgat1 gene.
What are your most exciting memories from the Department of Medical Genetics?
I must say I was very focused on my own work and a big race to publish! 1975 was a very demanding year! After that, cloning was beginning. There were other postdocs in the lab that got interesting data on new somatic cell mutants. What I really liked about being in Medical Genetics was that it was a very broad department, using genetic and biochemical strategies to understand things. So there were people like Mark Pearson working on muscle who were very mammalian, but there were also people working on phages, bacteria, yeast, and physical chemistry, like Jeremy Carver. Lou’s lab had a lot of connections with scientists at the OCI or Sick Kids, including Victor Ling, Larry Thompson, Manuel Buchwald and Bud Baker. So I had a really good education from many people who thought about science in a very broad way, and that was excellent training for me.
How did your experience in the Department of Medical Genetics influence your career trajectory?
It influenced my career a lot because somatic cell genetics became very topical in those days and people were looking for faculty with that type of expertise. Thus, I was invited to apply to Einstein because the Chair of Cell Biology Matthew Scharff was looking at genetic origins of the generation of antibody diversity using a somatic cell genetic approach, and was interested in having someone with that interest in his Department. Columbia University was also interested, as Larry Chasin, who was a somatic cell geneticist, was in the Biology department. New York was a good place as there were many options for both me and my husband. However, I liked Einstein in that it was very broad in its approach, had a philosophy of sharing equipment, generally being very supportive of new people and getting them going as fast as possible. So I came to Einstein, and so did my husband in the end, and we have both been here ever since.
What is your scientific discovery that you are most proud of?
I was very pleased that my lab was the first to inactivate the protein O-fucosyltransferase that is essential for Notch signaling in the mouse. Another group knocked out the same gene down in Drosophila a few months ahead of us, but we knocked it out in the mouse and showed that if you can’t put this particular fucose onto Notch, the mouse dies at mid-gestation. The phenotype of the mouse is very typical of a globally defective Notch signaling phenotype. My lab was also part of the exciting discovery of the sugar that is added after the fucose. So the fucose goes onto Notch and then after that goes another sugar, transferred by an enzyme called Fringe The Fringe gene was identified inDrosophila and so named because ectopic expression gives extra margins on the wings making a little fringe. The Fringe discovery was by a group of small labs, two Drosophilaand two mammalian. We published an article in Nature, in 2000 that presented a new paradigm for regulation of signaling that controls cell fate. Before that sugars were considered to be good for getting things to the right place, cell recognition or making glycoproteins stable or soluble, but the sugar addition by Fringe really directs Notch signaling at the right place, at the right time. Knocking out the fucose was then the next step and was published in 2003
How would you describe the major focus of your lab to a broad audience?
We are interested in defining, at a very precise level, what specific sugars do, and how they do it, in mammalian glycoprotein signaling receptors. These sugars are added to protein by enzymes and are chemically attached. The enzymes involved in attaching the sugars are often encoded by genes that have been conserved all the way through evolution, indicating conserved functions. There are many more-complicated sugars in bacteria and they do lots of things too, but in mammals we have a defined set of sugars added in different linkages that give rise to a lot of variation in structure. Why do we have all that variation in sugars? If you want to put a sialic acid onto something why not just have one enzyme? We don’t. We have 20. My lab is trying to define the factors that are required for putting important sugars onto Notch. We have two projects on Notch signaling. The first is on Fringe genes in mammals. There are three Fringe genes in mammals, but there is only one in the fly. Why do mammals have three? To find out we are focusing on T and B cell development in mutant mice. T and B cells become the lymphocytes that make antibodies and fight infection. It seems as though we need the three Fringe genes to have optimal T cell development and we want to understand why that is.
We also have a project on spermatogenesis. We showed that if we conditionally knockout MGAT1, the enzyme that makes a whole class of N-glycans, in spermatagonia of male mice all the spermatids fuse and that means you get no sperm, so those males are infertile. This is an interesting enzyme to investigate for design of an inhibitor as a male contraceptive, or even a rodent spermatocide. We are not focusing on that, but it is in the periphery of my mind. The next exciting thing that we discovered is a physoiologic inhibitor of this enzyme expressed in testis. We are trying to understand how the inhibitor influences spermatogenesis. We have the hypothesis that the MGAT1 enzyme and the inhibitor work together to help germ cells interact with Sertoli cells during the generation of sperm. The inhibitor may be modulating N-glycans to affect signaling or cell adhesion, and we are trying to work that out.
How have scientific collaborations shaped your research program?
Very much in that the discovery of GlcNAc-T1 was with Harry Schachter who had the expertise in the biochemistry. The Fringe activity discovery was with three other labs, and that was a great collaboration, very complementary. There have been about 40 – 50 people who have passed through my lab, and of course all that work is a collaboration. Without those collaborations, nothing would have happened. At Einstein we have had a number of people who have contributed and published with us. Basically, I am keen on collaboration. I seek it out. That in-depth expertise can be incredibly helpful. We do things ourselves by learning new techniques and we have excellent cores at Einstein. For example, we can knockout a gene in a mouse at Einstein without collaborating with someone. If it is going to push the work forward and the collaborator is going to have a similar level of motivation, then it is stimulating and fun to collaborate.
What do you think are the key elements of developing a successful research program?
I think including some possibility of serendipity is really important. If you select mutants in a way that is not designed to target a specific pathway but to get a phenotype, you will always get surprises. Now that we can target genes with CRISPR Cas9, you will still get surprising things because phenotypes will often not be what you expected. But finding really novel things I think requires designing experiments that will allow you to uncover something that you could never have imagined. I think building this component into your approach is important.
And then, having solid bread and butter projects for students and postdocs so that they can definitely get papers. Try for the high-flying stuff but make sure that the bread and butter gets published. It is important to keep your eye on publications. It is very hard these days because reviewers ask for a million additional experiments. Young faculty can get caught up in trying to publish in a high profile journal, spend a lot of time, and end up with one paper in 5 years, that may or may not, be in a high profile journal. It is really important to make sure that solid, good papers keep coming. The other stuff is icing on the cake.
Being generous with reagents is very important, as people will really appreciate it and you have nothing to lose and everything to gain. I do not co-author papers when I give reagents unless I make a significant contribution to the experiments, or in interpreting the data. A lot of people expect that if they ask you for a reagent the donor will want to be a co-author, but I always make it clear that it is not a requirement unless my lab is significantly involved.
Never react to reviews the day you receive them. Ever. Read them once. Put them away and then read them again next day.
When writing a serious e-mail, do not put the address in while writing the message, or write the message in a document that can be proofread first and not sent inadvertently.
With students and postdocs it is important to ensure that they get engaged. It is very rewarding to see them get excited with their project, if they are not at the beginning. For beginning faculty, it is important to have high standards for your lab people because if you don’t things can really go down the tubes. You can get a reputation for being strict, but too bad!
What advice would you give to graduate students hoping to pursue academic careers in science?
If you are thinking about an academic career in science you should talk to people who are in it and who enjoy it. There are a lot of problems these days with lack of funding and difficulty in publishing, and there can be a lot of negativity. I think if you like science you should give it the best go that you can and see what happens, because you might discover something unexpected like microRNA or who knows? If you are in it you can, if you are not in it, you can’t! I say go for it, and then you will have a nice PhD and a nice postdoc. My advice would be to be proactive and use the people around you, use the literature, take the ball, run with it, and be the leader of your project. If you take it as a personal mission to do the best with your project, I don’t see how you can go wrong unless you are very unlucky. There is usually something if you make sure that there is some bread and butter there. Even if your PhD is not what you hoped, it will probably still be good, and your postdoc might be brilliant. People mature at all different stages, and there is an element of chance. We have a scientist at Einstein who collaborated with others to discover the Ebola virus receptor, and lo and behold, it was a gene that another faculty member at Einstein had studied extensively and had all the mutant animals and reagents to put it quickly on the map. Neither of them knew that when the project started. The postdocs and students got a Nature paper that they could not have predicted. So that is why I say, hang in there, and as long as you are enjoying it, wait and see what happens.
What do you see as the biggest challenges facing scientists today?
It has never been so difficult to get a grant in my experience. Having a very thick skin is really important. You have to just keep writing grants. A lot of very good science is just not getting funded. I would say that between 25 and 50% of grants probably should be funded, because people have honed their points, have all their controls, made all the mice, and are ready to go. That would be the biggest challenge. It is also the biggest challenge facing institutions as they have relied on a historical model that is likely not going to last much longer. Young faculty usually do very well with getting their grants. It is the mid-level faculty that have a hard time with competitive renewals. We need to double or triple the budget for science so that the enormous possibilities of today can be realized.
The translational emphasis is another issue that is difficult. Clinical research is extremely expensive and usually asks defined questions using known reagents. It is rarely discovery science. It is getting to the moon but it is not finding out if there is another moon that we never knew about. So, I think that is a real problem for basic science. We need more support for basic science.