We were mainly focused on getting tissue. It seemed inevitable that HIPAA and U of M administrators would refuse our team access to Andrei's biopsy tissue. While distracted in this way, it's clear that we underestimated the magnitude of the difficulties we would face in actually culturing the tumor. The sample is finally obtained, cut into 18 little pieces and sent off to 3 separate institutions.
Once the cells were in hand, the polycellular, polymorphic nature of this tissue became clear. Cells were growing, but we could not be sure what they were. We were incessantly talking about how to obtain a pure culture of the tumor cells - how to get the normal fibroblasts and other contaminating cells separated from the grossly neoplastic ones. At the same time, the cells were showing an uncanny interdependency. The cells we thought were tumor cells (nicknamed 'puffer cells' for their fat little bodies and few, short, prickly projections) seemed to only thrive and cluster on top of the clearly normal fibroblasts. Immediately we started calling the fibrobalsts "nurse cells" and the puffer cells stem cells.
Lo and behold - the first sets of immunocytochemistry results came in, and one of the positive antigens is CD34 - a stem cell marker. Are these tumors really a rapidly expanding and proliferating population of stem cells? Then a real home run marker turns up positive - EGFR2 - the target of Herceptin. Again - it's a bit more clear why these cells seem to love fibroblasts. They must be getting needed growth factors from them, and luckily, one that they need is also one we can block. We have something to really bring back to the Oncology clinic and begin to target more effective therapy. Meanwhile, the cultures are slowly subdivided, and the nude mice just seem to be living their normal, hairless lives. Still, we lack the uniform, controllable wells of tumor cells that we need in order to test antimitotics and other treatments against. Also, we confirm Andrei's serum Thromboxane B2 level is again elevated about 10-fold over the population norm. This needs to be followed up as a possible diagnostic marker...
Frustration...At our request, the U of M Pathology lab re-tested for the EGFR2 (Her2-neu) antigen and came back with a negative result. We repeat our tests, not just on Andrei's tissue but on an array of control positive and negatives - and again confirm that Andrei's cells express an abundance of the Her2-neu protein. At the same time - the swarms of cancer cells in our cultures seem to be dwindling away, and the normal fibroblasts seem to be overtaking the cultures. We need to resolve the Her2-neu conflict - but how? The idea of using the monoclonal antibody in Herceptin as the actual immunocytochemical staining antibody would clear things up permanently - but we can't seem to get our hands on a bottle of Herceptin. Even a used up, spent dose vial from someone's therapy would have plenty of antibody for a stain. Can't someone in Oncology just retrieve one for us out of the trash? An intrigueing result is that our nude mice, who show no overt signs of a tumor burden, are showing elevated serum TXB2 concentrations. So, not only did Andrei show 10X elevated TXB2 a year before his radiologic diagnosis - the mice are also showing this when we can't see any tumor in them anywhere. We need to write up a short manuscript and publish this - we also need to get an archived sample of Andrei's serum from his remission period.
It has been a good period for Andrei, and we're all encouraged by this. He is back to work for many hours each week, and seems strong and unchanged from his normal, humble self. However, it has been a slow period for our research. We want to specifically thank Dr. Bloom and the clinical diagnostics lab at CLARiENT for his careful re-analysis of the H2-neu and his conference call to explain the diagnosis. Sadly for our team, we have had to give up on what we hoped would be a relatively effective and side-effect free biological treatment, as the amount of H2-neu making it to the surface of the SNUC cells is not sufficient to warrant any therapy. But on the positive side, H2-neu is a marker associated with poor prognosis in breast cancers. Maybe Andrei's currently promising response can be partly attributed to the lack of this mitogenic receptor. Meanwhile, the cell culture remains mired in the same formidable problem that it faced from the outset - how to get a homogeneous, purely neoplastic cell line out of the mixed cellular population of the biopsy tissue.
The cell population that we eventually screened out is certainly abnormal - it has multiple chromosomal abnormalities, and stains positively for CD34, intracellular H2neu, COX-1 and Thromboxane Synthase. The cells grow with agonizing slowness - to double the population takes 3-4 weeks instead of a more normal 3-4 days. They seem absolutely starved for an essential growth factor - or senescent - or both. Yet they are immortal - they don't die, they just drag along. Because of this, we are never able to obtain enough of them to kill them in any organized, scientifically controlled manner. Andrei is left to the treatment regimes familiar to the oncologists, and in the end, these fail him.
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| This page last changed on 22-Aug-2006 15:08 by snucnet. |