Chapter 04: Leaving MD Anderson for Industry: Research into Hybrid Proteins with Tyrosine Kinase Activity

Title

Chapter 04: Leaving MD Anderson for Industry: Research into Hybrid Proteins with Tyrosine Kinase Activity

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Dr. Arlinghaus explains that in 1983 he resigned from MD Anderson and took a job as Vaccine Development Director, Johnson and Johnson Biotechnology Center, in San Diego, California (1/1983-1/1986).They agreed to give him money to develop his CML research.He was able to secure his own laboratory space as a Visiting Investigator (Member) at the Scripps Clinic & Research Foundation in La Jolla (1/1983-1/1986).Dr. Arlinghaus describes his roles and notes that he arrived at Johnson and Johnson with an NCA grant awarded shortly before his departure from MD Anderson.Dr. Arlinghaus next describes the work he performed for Johnson and Johnson developing synthetic peptide vaccines.He describes the molecular chemistry that makes peptide vaccines impossible.He then outlines his next steps in studying the structure of the abnormal ABL protein in leukemia viruses.He describes the discovery of the "ABL-fused" gene in CML, adding a new dimension to his research and resulting in a 1995 paper.

Identifier

ArlinghausR_01_20140321_C04

Publication Date

3-21-2014

Publisher

The Making Cancer History® Voices Oral History Collection, The University of Texas MD Anderson Cancer Center

City

Houston, Texas

Topics Covered

The Interview Subject's Story - The Researcher; The Researcher; Evolution of Career; Professional Path; Overview; Discovery and Success; Definitions, Explanations, Translations; The Professional at Work; Understanding Cancer, the History of Science, Cancer Research; Finance, Entrepreneur, Biotechnology

Disciplines

History of Science, Technology, and Medicine | Oncology | Oral History

Transcript

Ralph B. Arlinghaus, PhD:

Well, because — because I looked like I was going to fail at MD Anderson, I took a job with Johnson & Johnson …

Ralph B. Arlinghaus, PhD:

… and helped start up a company in San Diego.

Tacey Ann Rosolowski, PhD:

Okay. This was in 1983.

Ralph B. Arlinghaus, PhD:

Yes.

Tacey Ann Rosolowski, PhD:

1983. Okay. Because, yeah, because I — I had noticed that you were away from the institution for three years.

Ralph B. Arlinghaus, PhD:

I went there because Johnson & Johnson agreed not only to — to have them — me help them develop their company, but allowed — but give me money to support my leukemia research.

Tacey Ann Rosolowski, PhD:

Now, can I ask you just a kind of a …

Ralph B. Arlinghaus, PhD:

So, in other words, I don’t take the job in La Jolla, in California, unless Johnson & Johnson agrees that they’re going to give me money to develop their company but also give me money to develop my research on chronic myeloid leukemia.

Tacey Ann Rosolowski, PhD:

Was this a, you know, kind of a leave of absence from MD Anderson?

Ralph B. Arlinghaus, PhD:

No, I resigned.

Tacey Ann Rosolowski, PhD:

You resigned. Okay.

Ralph B. Arlinghaus, PhD:

I was never coming back because, you know, I — I had — I had to move on to try to find out how to get — start working on chronic myeloid leukemia.

Tacey Ann Rosolowski, PhD:

What was it that — what made you make the decision to resign?

Ralph B. Arlinghaus, PhD:

I wasn’t getting money from NCI to fund my leukemia research so I said, well, I’ve got to get it from somewhere. So, I convinced J&J — they wanted me to develop their company. I said, well, I’ll come to California but only if you promise to give me money to support my research and allow me to attract trainees and pay them so that I can do that leukemia research at Johnson & in La Jolla.

Tacey Ann Rosolowski, PhD:

So, tell me about — first of all, what — what were you doing for them. Because I had down Vaccine Development Director.

Ralph B. Arlinghaus, PhD:

Well, they — there was a — a guy at Scripps Clinic named Richard Lerner.

Tacey Ann Rosolowski, PhD:

And then, you were a visiting investigator at Scripps Clinic and Research …

Ralph B. Arlinghaus, PhD:

I also had a …

Tacey Ann Rosolowski, PhD:

… Foundation.

Ralph B. Arlinghaus, PhD:

… NCI grant …

Tacey Ann Rosolowski, PhD:

Oh, okay.

Ralph B. Arlinghaus, PhD:

… so I worked …

Tacey Ann Rosolowski, PhD:

Mhmm.

Ralph B. Arlinghaus, PhD:

… I worked for Johnson & Johnson at their building in La Jolla. It was a rental facility.

Tacey Ann Rosolowski, PhD:

Mhmm.

Ralph B. Arlinghaus, PhD:

Then I worked at Scripps on my grant, working on one of these rapidly acting cancer genes called Moss ().

Tacey Ann Rosolowski, PhD:

Oh, okay.

Ralph B. Arlinghaus, PhD:

Because I didn’t know about the gene that caused CML.

Tacey Ann Rosolowski, PhD:

Mhmm.

Ralph B. Arlinghaus, PhD:

But — so I had a grant that I brought with me from MD Anderson. I did get funded but I got promised to have much more resources from J&J in addition to this grant from the National Cancer Institute, and that allowed me to hire people at J&J to help me investigate chronic myeloid leukemia.

Tacey Ann Rosolowski, PhD:

Mhmm.

Ralph B. Arlinghaus, PhD:

And there were physicians here – one guy named ____ who was working on chronic myeloid leukemia and he agreed to send me cell lines — cell lines is too strong — cells from chronic myeloid leukemia patients, packaged them up in dry ice, ___ frozen send them to La Jolla. He and some of his people came to La Jolla. I trained them how to do some of the assays so that they could do it here. But it was all J&J …

Tacey Ann Rosolowski, PhD:

All J&J money. Okay. Why did Johnson & Johnson agree to do that, do you think?

Ralph B. Arlinghaus, PhD:

Because they wanted to hire me to develop their company on synthetic peptide vaccines.

Tacey Ann Rosolowski, PhD:

Okay. So …

Ralph B. Arlinghaus, PhD:

And I was — my CV looked the best and they decided that I could — I could make Johnson & Johnson a lot of money to a peptide vaccine.

Tacey Ann Rosolowski, PhD:

What were these va — peptide vaccines supposed to do?

Ralph B. Arlinghaus, PhD:

Well, I’ll give you an example. Hepatitis B, serious human virus, causes hepatitis, which is liver disease and leads to liver cancer.

Tacey Ann Rosolowski, PhD:

And – I’m sorry, what - hepatitis C?

Ralph B. Arlinghaus, PhD:

Hepatitis B.

Tacey Ann Rosolowski, PhD:

Hepatitis B. Okay.

Ralph B. Arlinghaus, PhD:

Dick Lerner at Scripps got a — a bunch of money from J&J on this peptide vaccine concept.

Tacey Ann Rosolowski, PhD:

Mhmm.

Ralph B. Arlinghaus, PhD:

So, it was Lerner who convinced J&J to hire me because he knew about my work with Gag-Pol and all the proteins encoded for by leukemia. He knew about that exquisite work – exquisite may be too strong - …

Tacey Ann Rosolowski, PhD:

Sounded pretty exquisite to me.

Ralph B. Arlinghaus, PhD:

It — it was. And he knew — he knew me. He knew I was productive. It’s like Joe Schaeffer, when I got hired at MD Anderson, he told Felix Haas this guy, Arlinghaus, is going to make things happen. And I did. And when I went to California to work for J&J, Dick Lerner, who knew my papers, convinced them, 1) to hire me to come to California; 2) to work on his peptide vaccine. So, we hired seven or eight Ph.D. scientists to work on these peptide vaccine, for hepatitis, for influenza. None of that worked out. It was all failure. Not because of what I did; because there was a flaw that Lerner didn’t recognize. And, how am I going to explain that flaw to you?

Tacey Ann Rosolowski, PhD:

That’s going to be your challenge.

Ralph B. Arlinghaus, PhD:

When you inject a fragment of a protein called a peptide. Proteins – let’s say, hemoglobin in your cells, your liver cell and mine – they have a 12,000 molecular weight protein. There’s an alpha chain and a beta chain. They come together to form hemoglobin. That 12,000 molecular weight protein is a protein that has 120 amino acids. Lerner thought you could make a 10-amino acid fragment of globin, inject it into mice, rabbits, humans, and make a antibody that would neutralize hemoglobin.

Tacey Ann Rosolowski, PhD:

Right.

Ralph B. Arlinghaus, PhD:

Although he didn’t talk about hemoglobin. He was doing it with viral proteins. But, the point is he — he had this concept that if you had a small chunk of viral protein and you made it chemically in a lab, that if you immunize an animal, that small chunk of protein would make an antibody which would then recognize proteins made or secreted by the virus …

Tacey Ann Rosolowski, PhD:

Right.

Ralph B. Arlinghaus, PhD:

.. and it would be useful to neutralize the virus. But, it turns out what Lerner failed to recognize, I didn’t know, I learned it for him and told him about it, is that these peptide vaccines — I can’t use more words you don’t know of – had limited — if it’s a 10-amino acid peptide compared to a 120-amino acid protein, the number of sites that an antibody would tightly bind to would be three or four for the 10-amino acid peptide. But it would be ten times that for the whole protein. And, it turns out that the affinity of antibodies for their proteins requires multiple binding sites in the target protein. And peptides didn’t identify those other – let’s say, three – 25 different binding sites. They only identified a few so your antibodies only detected a small portion of the viral protein.

Tacey Ann Rosolowski, PhD:

That’s a great explanation …

Ralph B. Arlinghaus, PhD:

And it didn’t bind very tightly …

Tacey Ann Rosolowski, PhD:

Yeah.

Ralph B. Arlinghaus, PhD:

… so it was — what we call low affinity. So it turned out there were no peptide vaccines, we didn’t make one. No one else made one. Other people tried. That’s failed technology because of not understanding that you couldn’t use short peptides as antigens to — to – what shall I call it – to — to – looking for a word – you couldn’t use short peptides that may represent 5% of the amino acids of the total protein to accomplish, to allow the immune system to decorate all the sites. It would only to decorate those sites that were in the short peptides. And you needed all the sites to be decorated by antibodies.

Tacey Ann Rosolowski, PhD:

Makes sense. Good explanation. Thank you.

Ralph B. Arlinghaus, PhD:

And we didn’t know it back then.

Tacey Ann Rosolowski, PhD:

Yeah, you didn’t know it. But it seems like you had to go thro — go through that process …

Ralph B. Arlinghaus, PhD:

That’s right.

Tacey Ann Rosolowski, PhD:

… to figure it out.

Ralph B. Arlinghaus, PhD:

That’s right. Because what we got were antibodies that recognized the peptide, but antibodies that only poorly recognized the viral protein.

Tacey Ann Rosolowski, PhD:

1 So, that’s what you were doing for Johnson & Johnson …

Ralph B. Arlinghaus, PhD:

Johnson & Johnson.

Tacey Ann Rosolowski, PhD:

… so what were you doing in your CML …

Ralph B. Arlinghaus, PhD:

Well, when — when I moved to Johnson & Johnson, a paper by now a colleague called John Groffen – G-r-o-f-f-e-n, John Groffen and his wife, Nora Heisterkamp, published a paper that said that – thinking of the words – that — they were studying the structure of this abnormal ABL protein present in cells like Rauscher leukemia virus and — and Moloney leukemia virus, and now in patients with CML. What they found was that there was a gene called ABL, this genome, reading this mini review about that’s fused. Remember CML is a disease where you’ve got part of the ABL gene fused to part of the BCR gene. And so, Groffen found out that — that hybrid chromosome produced a hybrid messenger RNA made up of sequences from BCR and ABL gene, and that — and then to produce a protein that was made up of ABL sequences and BCR sequences. So, it was a BCR-ABL protein and that protein was made in every CML patient that he examined. So, I wrote ____ and asked him to send me cell lines or cells from patients with CML to see if I could detect that abnormal ABL protein using sophisticated peptide technology that I learned about from Lerner, make short peptides, make antibodies, and those antibodies would see that short peptide and would see it to some degree in the protein itself. So — so, I used that technology to identify and publish a paper that in CML cells, there was a protein that had sequences from the ABL protein and sequences from the BCR protein, leading to the conclusion that it was a BCR-ABL protein. And more than that, identify that BCR-ABL protein as a tyrosine kinase that had activity in and of itself, that was always active in itself. So that finding was made possible by Groffen and his wife, Nora Heisterkamp.

Tacey Ann Rosolowski, PhD:

And her name, Hiserkamp?

Ralph B. Arlinghaus, PhD:

Heisterkamp

Tacey Ann Rosolowski, PhD:

Heister — H-e-i- …

Ralph B. Arlinghaus, PhD:

H-e-i-s-t-e-r-k-a-m-p.

Tacey Ann Rosolowski, PhD:

… k-a-m-p. Okay, thank you.

Ralph B. Arlinghaus, PhD:

Nora Heisterkamp.

Tacey Ann Rosolowski, PhD:

Nora. Great. Alright, thank you.

Ralph B. Arlinghaus, PhD:

So, in any event, all the science we all do in our field, what — whatever we’re working on, it’s all built on discoveries from other people.

Tacey Ann Rosolowski, PhD:

Sure. Now, this — I mean, leading up to the tyrosine kinase activity, because that, as I learned from background research for other folks that I’ve — I’ve prepared to interview. I mean, that — that’s a word that’s come up in terms of signaling pathways and crosstalk ….

Ralph B. Arlinghaus, PhD:

That’s right.

Tacey Ann Rosolowski, PhD:

… and all the receptor sites.

Ralph B. Arlinghaus, PhD:

That’s right.

Tacey Ann Rosolowski, PhD:

So, is that adding a new dimension to your work at this point, or …

Ralph B. Arlinghaus, PhD:

Yes.

Tacey Ann Rosolowski, PhD:

… were you ___

Ralph B. Arlinghaus, PhD:

Yes.

Tacey Ann Rosolowski, PhD:

Okay. So this added an entirely new …

Ralph B. Arlinghaus, PhD:

I knew that the BCR-ABL hybrid protein was a tyrosine protein kinase. That means it — it bound ATP and took the phosphate from ATP and put it on target proteins.

Tacey Ann Rosolowski, PhD:

Now, could you tell me now, given what you’ve learned since then, what is the role of those protein products in that transfer of phosphates, what’s the role of that in cancer?

Ralph B. Arlinghaus, PhD:

Well, I mentioned this two-faced god called Janus. Well, as — as a — as a young assistant professor at MD Anderson, I recognized what was known in the field by a guy named Jim Iley, again standing on the shoulders of — of these great giants. Jim Iley found out, along with many others 25 years ago, that Janus kinase is — plays a very important in blood cells for making more blood cells and making blood cell proteins. So, I reasoned that the BCR-ABL oncoprotein causing leukemia in blood cells — affecting blood cells would probably partner with Janus kinase, and that — that guess in 1995 turns out to be correct in 2011. So, my — my papers have led to this scenario. That the Philadelphia chromosome forms, we don’t know how, by accident, fusing parts of BCR and parts of ABL to give you this hybrid, this has a eternally active tyrosine kinase from the active ABL sequences in the BCR-ABL. And, now we know that that sequence, that kinase, is one of the things it does is activate, turn on Janus kinase 2 in CML cells. And then, we published that Janus kinase 2 activates the RAS and PI 3 kinase pathways in leukemia.

Tacey Ann Rosolowski, PhD:

I’m sorry, P — P …

Ralph B. Arlinghaus, PhD:

RAS, R-A-S…

Tacey Ann Rosolowski, PhD:

Mhmm, and …

Ralph B. Arlinghaus, PhD:

… and PI 3 kinase pathways in leukemia cells. And use — people used to think was BCR-ABL kinase that did that but it was actually Janus kinase that did it because BCR-ABL kinase was necessary to activate Janus kinase so it could do that.

Tacey Ann Rosolowski, PhD:

And so, these are all pathways that are essential.

Ralph B. Arlinghaus, PhD:

Quarter after two. Okay.

Tacey Ann Rosolowski, PhD:

We’re good?

Ralph B. Arlinghaus, PhD:

We’re good.

Tacey Ann Rosolowski, PhD:

Okay.

Ralph B. Arlinghaus, PhD:

So far.

Tacey Ann Rosolowski, PhD:

Okay.

Ralph B. Arlinghaus, PhD:

Forty-five minutes. I’m talking too damn much.

Tacey Ann Rosolowski, PhD:

No, that’s fine. We have another session scheduled so, you know, we’re — we’re good.

Ralph B. Arlinghaus, PhD:

Okay, alright.

Tacey Ann Rosolowski, PhD:

It — it will take whatever time it takes. No problem.

Ralph B. Arlinghaus, PhD:

Yeah.

Tacey Ann Rosolowski, PhD:

So, am I correct in understanding that these — these different functions, you know, one step to the next step, to this next step, to this next step …

Ralph B. Arlinghaus, PhD:

Yes.

Tacey Ann Rosolowski, PhD:

… are all involved in keeping that leukemia cell active …

Ralph B. Arlinghaus, PhD:

Yes.

Tacey Ann Rosolowski, PhD:

… keeping it dividing, keeping it …

Ralph B. Arlinghaus, PhD:

Keeping it alive.

Tacey Ann Rosolowski, PhD:

Keeping it alive. Okay. So, they’re all — it’s basically part of the, you know, life function, if you will, …

Ralph B. Arlinghaus, PhD:

It is.

Tacey Ann Rosolowski, PhD:

… of the cancer cell. So, do you — were you able to discover precisely what roles these serve inside the cancer cell?

Ralph B. Arlinghaus, PhD:

Others have done that. Like the RAS pathway …

Tacey Ann Rosolowski, PhD:

Okay.

Ralph B. Arlinghaus, PhD:

… and what it does in normal cells as well as cancer cells. That was done by others.

Tacey Ann Rosolowski, PhD:

Okay.

Ralph B. Arlinghaus, PhD:

The PI 3 ki — I just learned how CML cells, which were known to have activated RAS pathway, activated PI 3 kinase pathway — I learned how that came about. That’s because BCR-ABL activated JAK 2 …

Tacey Ann Rosolowski, PhD:

Mhmm.

Ralph B. Arlinghaus, PhD:

… and then JAK 2 phosphorylated a site on tyrosine that led to activation of the RAS and PI 3 kinase pathways. That’s what’s in that leukemia paper in 2011.

Tacey Ann Rosolowski, PhD:

Interesting.

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Chapter 04: Leaving MD Anderson for Industry: Research into Hybrid Proteins with Tyrosine Kinase Activity

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