from John
Besides the Eureka moment of making an important or, at least, useful discovery the most satisfying experience I had in research was receiving a fundable rating on my federal grant applications from other scientists. In this process, notable scientists would sit in committees in Washington DC, called study sections, and discuss the merits of hundreds of grant applications and then vote on them resulting in scores that were above or below the fundable level. On a few occasions I was invited to become an ad hoc member of such committees because of the nature of my expertise. The next most satisfying experience was having a research paper accepted for publication; this also happened as a result of peer review; and I published about 60 research papers during my career – the more prestigious the journal, the greater the feeling of accomplishment. The next most satisfying event was seeing others pick up on my findings and extend them to something more valuable. This form of satisfaction usually comes several years later. I checked the citations of my papers recently and found that some of my papers are still being cited as references some 15 to 30 years after the papers were published.
One protein that I discovered and named plastin continues to be investigated as a biomarker and possible drug target for diagnosing and treating various forms of cancer. In 1978 I performed an experiment that led to the discovery of plastin and another interesting protein. This led to NIH funding of my research program for 13 continuous years after I left NIH several years later. In 1985, I submitted a paper to Cancer Research and the reviews came back positive, but one reviewer suggested that I give the protein that was the subject of the paper a name. I was tickled by the idea of actually naming a human protein (and its gene).
I had a concept for the function of the protein because of its normal expression in all white blood cells and its mysterious appearance in tumor cells from solid tissues (carcinomas and sarcomas). 
I thought that it may make adherent cells that are sort of flat behave more like white blood cells that are sort of round and able to migrate throughout the body through the circulatory system and interstitial space.
In cancer biology the migration of a carcinoma cell to other tissues – like breast cancer spreading to the lymph nodes – is called “metastasis”. It was my suspicion that this protein gave adherent cells more plasticity in their shape and motility so I named the protein “plastin”. I got the idea for the name from the movie The Graduate which was a big hit in 1967.
Remember that scene…
When we cloned the gene for plastin, we discovered in the process that there were two slightly different plastins, one found only in white blood cells (L-plastin) and one found only in solid tissues (T-plastin). Today both isoforms of plastin are being investigated for their roles in tissue adhesion, cell motility, cell architecture, their respective roles in different forms of cancer. I was really surprised when one researcher on the West Coast discovered that T-plastin was inappropriately expressed in Sezary lymphomas which derive from T-lymphocytes – a white blood cell involved in immune and inflammatory responses due to infections and tissue damage. Sezary cell lymphoma is often lethal. This lymphoma is distinct from other forms of lymphoma and leukemia because it invades the skin and behaves sort of like a solid tumor on the surface of the skin. As the result of this finding there is now an approach to making drugs that could conceivably lead to a treatment for Sezary lymphoma by targeting T-plastin. This new technology which was first published in 2001 is called “interfering RNA” or siRNA for “short interfering RNA”. It would be hard to fit an explanation of how siRNA works as a drug into this story so I won’t try.
In April I began covering a major lawsuit at a website called RNAiLitigation . The lawsuit was filed in June 2009 by the prestigious German university system known as the Max-Planck Institute against MIT’s prestigious Whitehead Institute and the University of Massachusetts. This litigation is a fight over who has the right to use and sell the siRNA invention previously assumed to be owned by Max-Planck. I had to read carefully the seminal papers and patents on this invention to prepare myself for telling the story behind the lawsuit and covering the testimony and developments in the upcoming trial. When I read the seminal paper by Thomas Tuschl on his discovery of siRNA and how it works, I found that the paper was co-authored by Klaus Weber a highly revered long-time professor at Max-Planck in Goettingen. I was somewhat pleased to see that Klaus was still around and doing great things in science, especially because we did something great together back in 1980.
I knew of Klaus Weber and his wife Mary Osborne long before I ever conceived of contacting him in 1979. Weber and Osborne were famous for their work at Harvard which I won’t go into here. I used a technique they developed in the late 1960s when I was a grad student working on my Ph.D. In the mid 1970s, Klaus received a professorship at Max-Planck and developed his well-known research program investigating the nature of mammalian cellular architecture and motility. His lab had published the amino acid sequences of muscle actins from various species down to single cell eukaryotes (cells with nuclei). In 1978 on that day when I discovered plastin, I also discovered the first mutation event in a human actin called beta-actin that along with gamma-actin was the most abundant architectural protein of all non-muscle cell types. I wanted to determine the sequence of this mutant actin to prove that a mutation had occurred. This required expertise that was out of the scope of my own area of research and I was advised by a specialist in this field who work downstairs from me at NIH. He suggested that the only way to get this done was to find a laboratory that specialized in sequencing actins. There was only one in the world, Klaus Weber’s lab at the Max-Planck.
Back in 1979 the only way to communicate what I had found effectively was to write Klaus a letter. Two weeks later I received a letter back saying that he and his post-doc Joel Vandekerckhove would be willing to sequence my mutant actin and he told me how I should prepare the cell extract containing the mutant actin so that they could purify it and sequence it. The speed of Klaus’ response told me that he was seriously interested. Years later after more experiences like this, I had learned many times over that when you collaborate with sharp people, extraordinary things happen. In December of 1980 we published the complete amino acid sequences of the normal and mutant human beta-actins and the other gamma-actin in the top biomedical journal Cell. The mutant beta actin had incurred a single amino acid exchange that allowed us to predict the actual mutation in the gene sequence.
In the summer of 1981 before I moved to Palo Alto CA, Klaus called me up and proposed that we meet for the first time at the Shoreham Hotel just north of Rock Creek Park in DC just down the road from NIH in Bethesda. So we got together one morning in the lobby of the hotel. I had made a lot of monoclonal antibodies against human architectural proteins (this architectural structure is called the “cytoskeleton”) as a by-product of something else I was doing. I had little use for these antibodies and shipped them off to Goettingen since Klaus was well known for using immune antisera to visualize the cellular cytoskeleton. The method for making epitope-specific monoclonal antibodies was new and this had not been tried in Klaus’ lab. It turned out that he wanted to show me some pictures of cells stained with antibodies that I had sent him. I think that he was also trying to find out what I intended doing with them which was a proper thing to do. The photos were very impressive and suggested that these antibodies could be used to map the cytoarchitecture of all cell types. He asked me vaguely “What are you going to do next?” I told him that my only interest was in cloning the human actin genes to reassure him that he could move forward with the monoclonal antibodies. That was our last communication until April 2010.
As I mentioned above, Klaus was a co-author on the seminal paper describing the invention of siRNA in 2001 – still today the most exciting biomedical discovery since the mid-1980s. I had notified the Technology Transfer Office for the Max-Planck in Munich about our coverage of the litigation they were involved in. I had mentioned Klaus Weber’s involvement in the discovery of siRNA at the website.
In April I received an email from “Mary Osborne”. It turned out to be Klaus using his wife’s email much in the same way I use Becki’s email. I had said at the website that “It’s is a small world” and he said it that “indeed it was a small world” and explained to me his simple contribution to Tuschl’s discovery of siRNA. He also said that he remembered our exchanges. Needless to say, I was thrilled to hear from him this one last time.
I went back to that seminal paper to look more closely at his ‘small’ contribution from his own description. When I looked at the particular experiment that was used to demonstrate that Tuschl’s invention worked, I realized a lasting connection. When we met at the Shoreham Hotel nineteen years earlier, one of the antibodies that I had provided to him stained the nuclear envelope, a protein called “lamin”. I think before showing me the pictures, Klaus had already realized the power of mono-specific monoclonal antibodies. His lab proceeded to make them and use them extensively in his research. Although not the very same antibody that I had prepared, his anti-lamin antibody was the one that was used elegantly to demonstrate shutdown of lamin expression with siRNA targeting the lamin messenger RNA, e.g. gene silencing. This entire story is emblematic of how discovery science works.
In the ensuing nine years that seminal siRNA paper has received close to 6000 citations from other research papers, and siRNA-based drugs are now progressing through the early stages of clinical development.
