Defining Nanoproteomic Profiles of Hypoxia Signaling to Diagnose Clear Cell Renal Cell Carcinoma
Abstract/Contents
- Abstract
- The loss of function of the von-Hippel-Lindau (VHL) tumor suppressor gene leads to the development of clear cell Renal Cell Carcinoma (ccRCC) through activation of hypoxia signaling. ERK 1/2 is an important protein downstream of many hypoxia pathways, known to play a critical role in ccRCC disease progression. ERK activation occurs through phosphorylation. Previous studies have shown that total ERK 1/2 and phosphorylated ERK1/2 expression levels are higher in RCC cells compared to normal cells. However, ERK1/2 has two phosphorylation sites, and standard methods cannot distinguish between its different phosphorylated states. We hypothesized that ccRCC cells might exhibit selectivity for specific phosphorylated isoforms of ERK (such as mono-phospho-ERK1, denoted as pERK1). Further, we hypothesized that the relative percentage of ERK1 and ERK2 phospho-isoforms (pERK1, ppERK1, pERK2, ppERK2) can distinguish ccRCC tumor cells from adjacent normal renal parenchyma. Using a high-throughput nanoscale immunoassay (NIA), we measured the percent phosphorylation of ERK 1/2 and its respective isoforms within tumor and adjacent normal tissue in 20 ccRCC patients. For the first time, we showed that there was a significant difference in the percentage phosphorylation of each of the ERK 1/2 phospho-isoforms between ccRCC tumor and normal renal tissue. Through paired analysis, we found that the percentage phosphorylation of both mono-phosphorylated ERK1 and dual-phosphorylated ERK1 were higher in tumor tissue versus normal tissue (p<0.002 and p<0.00004, respectively). Similar results were obtained for mono-phosphorylated ERK2 and dual-phosphorylated ERK2 (p<0.01, for both). Thus, these results demonstrate that quantifying percent phosphorylation of key signaling molecules might be useful in defining signatures of ccRCC.
Description
Type of resource | text |
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Date created | June 15, 2014 |
Creators/Contributors
Author | Praharaj, Dave | |
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Degree granting institution | Stanford University, Department of Biology, 2014 | |
Primary advisor | Felsher, Dean | |
Advisor | Gozani, Or |
Subjects
Subject | clear cell Renal Cell Carcinoma |
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Subject | Nanoscale immunoassay |
Subject | Hypoxia signaling |
Subject | ERK 1/2 |
Genre | Thesis |
Bibliographic information
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- User agrees that, where applicable, content will not be used to identify or to otherwise infringe the privacy or confidentiality rights of individuals. Content distributed via the Stanford Digital Repository may be subject to additional license and use restrictions applied by the depositor.
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- This work is licensed under a Creative Commons Attribution Share Alike 3.0 Unported license (CC BY-SA).
Preferred citation
- Preferred Citation
- Praharaj, Dave and Felsher, Dean and Gozani, Or. (2014). Defining Nanoproteomic Profiles of Hypoxia Signaling to Diagnose Clear Cell Renal Cell Carcinoma. Stanford Digital Repository. Available at: http://purl.stanford.edu/bw243zz1996
Collection
Undergraduate Theses, Department of Biology, 2013-2014
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- Contact
- praharaj@stanford.edu
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