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DISCUSSION
Dr Marybeth Hughes (Bethesda, MD): What I was wondering was, you said at the very end that the KCC4 was associated with decreased functional status. In my experience, they tend to be about 50- 50 functional, and I noticed that 17 of your 26 were overamplified in KCC4. Can you tell me the breakdown of functional versus nonfunctional in your sample population?
Dr Taylor C. Brown: We had 32 total samples. We were unable to do the copy number assay in all the samples, but of those 32 samples, 11 were nonfunctional.
Dr Michael J. Demeure (Phoenix, AZ): I have a couple of disclosures. I am the Executive Vice Pres- ident and General Manager of Ashion Analytics, and I am a consultant for Tekmira Pharmaceuti- cals. Your work is very nice. I like the fact that you showed that overexpression correlates with amplification. Did you correlate with IGF1, which is a regulatory gene, and we heard IGF2 is impli- cated but there’s some crosstalk. In other tumors, KCC4 has been correlated with poor survival. I think it’s been looked at in breast and cervical can- cer and the like. Can you comment on the clinical outcome as well?
Dr Taylor C. Brown: Yes, KCC4 overexpression has been associated with poor clinical outcomes in breast, ovarian, and cervical cancer. Those all have been recently demonstrated. With respect to correlation with IGF levels, we did measure IGF levels, mRNA levels, but we found overexpression in essentially all samples, so it was hard to correlate overexpression of all samples with some samples that are decreased or increased for KCC4.
Dr Geeta Lal (Iowa City, IA): No disclosures. I just have a couple of follow-up questions. I realize they’re not a huge number of patients, but was there correlation with greater stage, vascular inva- sion, any other clinical features? Second, I guess it’s hard to do that in the primary patient samples, but have you looked at cell lines. Does it function the same way in adrenal tumors as it does in other tumors? Does it affect invasion? Does it affect migration?
Dr Taylor C. Brown: We did look at clinical char- acteristics. The only association we found was with nonfunctional tumors, which makes some sense, because channel dysregulation has been shown to
play a prominent role in adrenal tumors. It did not correlate, however, with stage or survival, I think in part because we had so few samples, and we didn’t have enough. In terms of functional studies in cell lines, we have performed those studies. We have some preliminary results that do show, nonpeer- reviewed studies that do show that KCC overexpression in ACC cell line SW13 does promote cell migration and invasion.
Dr Douglas Evans (Milwaukee, WI): I just wanted to follow up on that last question. Because the histopathology is complicated in this disease, are you sure that the patients without overexpres- sion actually had adrenocortical carcinoma? Did you look at their clinical outcome?
Dr Taylor C. Brown: Yes. All these samples, before they are tested, are reviewed by a dedi- cated endocrine pathologist at our hospital. As far as we can tell, on review of the medical record, these patients all suffered from adrenocortical malignancy.
Dr Douglas Evans: I think it’s a problem, then, if you have patients who died of ACC and they didn’t
have overexpression, why would you pursue this pathway?
Dr Taylor C. Brown: ACC is an incredibly het- erogeneous tumor. We know that from our own next-generation sequencing results. One tumor can be very different from another tumor. By no means do I think this is a uniform mechanism that plays out in every tumor, but I think it’s impor- tant to understand any potential molecular targets that do occur in ACC.
As we all know, the treatment modalities are limited at this point. Targeted therapies to date have had some efficacy, but it has been limited.
Dr Emad Kandil (New Orleans, LA): Did you also look at epidermal growth factor in relation to the high copy numbers of KCC4? Also, did you examine any epithelial or mesenchymal markers to prove the role of KCC4 in a terminal progression?
Dr Taylor C. Brown: No, we did not look at epidermal growth factor. We’ve thought about it, but we haven’t done it. We have looked at some cadherins, but we are struggling with those studies.
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