PRMRP 2012 Investigator Vignette

Title: Nanoscale Proteomic Analysis of Oncoproteins in Hematopoietic Cancers

Investigator: Dean Felsher, MD, PhD, Stanford University

So the objective of our DoD grant proposal was to develop a new technology that would allow us in real-time to measure the therapeutic response of drugs for cancer. And, what we specifically wanted to do, was be able to measure proteins and changes in proteins that were the targets of specific therapeutics.

So, my research program is devoted to figuring out how oncogenes cause cancer and whether or not shutting down oncogenes could be used to treat cancer. And what we reasoned is that, there will be some cases where if you turn off that oncogene, the cancer will completely go away. And the idea that this could happen is coined "oncogene addiction." This has been referred to as an Achilles heel of cancer.

But in order to do that and make therapies, you need to be able to have drugs that target oncogenes, and be able to measure whether or not that particular therapy is actually able to turn off that oncogene. And you'd like to be able to do this in many different points during patients' care. So we developed a new technology called the nanoscale immuno-assay for exactly this purpose. And the technology allows you to use very, very small amounts of material-25 cells, 4 nanoliters of material-then that material is homogenized and passed through a capillary tube, and the different proteins including the oncoproteins, are now separated based on their charge. And then using antibodies, we can actually detect very specific proteins we're interested in, like the MIC oncoprotein and the BCL2 oncoprotein, and the technology is highly sensitive and very, very quantitative.

And so our reasoning was, with this super powerful technology, we should be able to then measure in a patient, before and after they get a specific therapy, a response in their proteins, a phosphorylation response. The tumor would have a specific pattern, and in response to therapy that pattern would change for a particular oncoprotein that will tell you whether or not the drug was actually working in the patient.

We had a clinical study where we were giving a drug called Atorvastatin to patients with lymphoma. And we put a needle into their lymphoma before they received this drug, and we put a needle into the lymphoma 8 days after they had received this drug, and we measured two oncoproteins, STAT5 and STAT3. Using this technology, we can measure a phosphorylation of STAT5 and STAT3 very quantitatively, exactly where expected, and in response to this drug you can see that now you no longer detect the phosphorylated versions of these proteins.

Now this technology is very sensitive at detecting patterns that distinguish also cancer cells from normal cells. And this shows an example in a solid tumor, where we measured an oncoprotein called ERK. And you can see that there's these distinct phosphorylated forms of ERK. They're not very abundant but they're present. But if you look in the adjacent normal tissue, you do not see any of these phosphorylated forms. So this potentially could be used as a way of telling whether or not a tumor mass is a cancer versus just normal tissue. And this shows the quantification of a specific phosphorylated form of ERK, very high in tumor, extremely low in non-tumor.

So, this nanoscale technology allows us to predict therapies that are useful to treat cancer; it can also be used to distinguish normal cells from cancer cells, and it allows you in real-time to measure how therapies are working, and this enables you to detect very quickly after giving a therapeutic whether or not a drug is having its intended benefit. We're now applying this technology in many other clinical circumstances. We initially proposed in our work for the DoD to look at hematologic cancers, but we're now looking at many different solid tumors.

What's also exciting is the work we've done has been so interesting to other colleagues at Stanford, they're using this same technology to measure therapeutic response not just in cancer, but also in many other diseases, such as heart disease and diabetes and infectious disease. All of this was stimulated from one grant that came from the DoD.