2013hakimiaphd.pdf (6.78 MB)
Plasma Protein Profiling for Bladder Cancer Biomarker Discovery using UPLC-HDMS[superscript E ] Label-Free Quantitation
thesisposted on 2013-10-08, 12:03 authored by Amirmansoor Hakimi
In the UK, bladder cancer is the 4th most common cancer in men and 11th most common in women. In 2010, just over 10,000 new cases were diagnosed and 4,900 deaths were recorded. At their first diagnosis, the majority of bladder cancer patients (75-85%) present with non-muscle invasive disease. In 50-70% of these patients the tumour will recur and in 10-20% of them it will progress to muscle invasive disease. Mass spectrometry based proteomics has been chosen for clinical biomarker discovery due to its ability to perform qualitative and quantitative protein profiling on clinical samples. In total 90 plasma samples were used in this study in two groups of disease and control. An optimised and evaluated UPLC-IMS-DIA-MS[superscript E] label-free quantitation method was used for plasma protein profiling. To our knowledge, this is the first report investigating the biomarkers of bladder cancer incorporating label-free quantitation and UPLC-IMS-DIA-MS[superscript E] methodology. To assess expression level of proteins of samples in different groups a plan consisting of four data processing packages was used. Each of the packages uses different statistical means by which to identify proteins and/or compare expression levels alteration. Optimisation of the methodology helped in the thorough investigation of the plasma proteome with coverage of up to five orders of magnitude of plasma protein concentration dynamic range. In total, 11 proteins were found as possible markers of diagnosis for bladder cancer. Four of these candidates (afamin, alpha 1-B-glycoprotein, apolipoprotein-A1 and haptoglobin) were previously reported to be urinary markers of bladder cancer. CRP was overexpressed when plasma samples from patients with low grade-Ta tumours were compared to every other sample and may be used as a diagnostic marker. Similarly, afamin and haptoglobin were overexpressed in plasma samples from patients with high grade-high stage tumours when compared to samples from patients with high grade-low stage disease.
Supervisor(s)Jones, Don J.L.; Jones, George D.D.
Date of award2013-10-01
Awarding institutionUniversity of Leicester