There is an urgent need for new blood biomarkers for epithelial ovarian cancer (EOC), which is the most lethal gynecological cancer in the United States. CA125 is the most commonly used FDA approved biomarker for EOC, but it is not approved for early diagnosis due to inadequate sensitivity and specificity. Furthermore, although CA125 is commonly used to assist in clinical management of the disease after diagnosis, it cannot be used for the substantial portion of EOC patients where CA125 is not substantially elevated at time of diagnosis, which includes the clear cell and mucinous subtypes. Our long-term goal is to identify novel EOC blood-based and tumor tissue biomarkers and conduct laboratory-scale validation to determine which biomarkers can improve early diagnosis and/or clinical management of this disease. Our overall working hypothesis is that the most specific EOC blood biomarkers will be proteins shed by the ovarian tumor into the blood and that are at very low abundance levels in the blood of cancer-free individuals. During the past grant year, we achieved all goals of the original proposal. Specifically, we identified 29 low-abundance, novel biomarkers using a novel in-depth proteome analysis method to identify human proteins in serum or plasma of xenografted mice containing human tumors. These proteins were then shown to be elevated in blood from EOC patients compared with non-cancer controls using a label-free multiplexed multiple reaction monitoring (MRM) mass spectrometry (MS) assay. We have now narrowed this list to 12 high priority biomarkers using biological and functional criteria Our specific goal for the next grant period is to conduct laboratory-scale validation of these 12 biomarkers in several larger patient cohorts to identify the best biomarkers for early diagnosis of EOC and for improved clinical management of this disease. We will also conduct preliminary evaluations of the capacities of these biomarkers to distinguish among major EOC subtypes, as these are genetically different diseases. To determine whether some biomarkers could complement CA125, we will conduct a focused evaluation of secretomes of clear cell and mucinous ovarian tumors. We will also separately evaluate the predictive value of our biomarkers in the subset of patient plasma samples that exhibit low CA125 levels in our new larger patient cohorts. The two aims that will be pursued over a 5- year period are: 1) Develop tissue and high-throughput preclinical plasma assays for our 12 novel EOC biomarkers, and 2) Validate sensitivity and specificity of our 12 novel EOC biomarker levels in plasma or serum from multiple independent patient cohorts. The biomarkers will be quantified in patient and control plasma or serum using a novel ImmunoMRM method that multiplexes pull-down of targeted intact proteins by commercially-available antibodies using stable isotope-labeled intact proteins as internal standards. It is expected that the subset of our 12 biomarkers that are shown to be useful for early diagnosis and/or clinical management of EOC will advance to larger-scale multi-center, preclinical testing in CLIA certified laboratories in the future.

Public Health Relevance

New blood biomarkers for early detection of epithelial ovarian cancer and to assist in clinical management of the disease are urgently needed because ovarian cancer is a major cause of death from cancer in women. We recently identified a substantial number of very promising new ovarian cancer biomarkers that now need to be tested in multiple larger groups of ovarian cancer patients to determine which biomarkers have the highest potential to improve patient health and survival.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA131582-05A1
Application #
8759302
Study Section
Cancer Biomarkers Study Section (CBSS)
Program Officer
Patriotis, Christos F
Project Start
2007-12-01
Project End
2019-06-30
Budget Start
2014-08-11
Budget End
2015-06-30
Support Year
5
Fiscal Year
2014
Total Cost
$440,659
Indirect Cost
$202,465
Name
Wistar Institute
Department
Type
DUNS #
075524595
City
Philadelphia
State
PA
Country
United States
Zip Code
19104
Beer, Lynn A; Wang, Huan; Tang, Hsin-Yao et al. (2013) Identification of multiple novel protein biomarkers shed by human serous ovarian tumors into the blood of immunocompromised mice and verified in patient sera. PLoS One 8:e60129
Tang, Hsin-Yao; Beer, Lynn A; Tanyi, Janos L et al. (2013) Protein isoform-specific validation defines multiple chloride intracellular channel and tropomyosin isoforms as serological biomarkers of ovarian cancer. J Proteomics 89:165-78
Tang, Hsin-Yao; Beer, Lynn A; Chang-Wong, Tony et al. (2012) A xenograft mouse model coupled with in-depth plasma proteome analysis facilitates identification of novel serum biomarkers for human ovarian cancer. J Proteome Res 11:678-91
Cao, Zhijun; Tang, Hsin-Yao; Wang, Huan et al. (2012) Systematic comparison of fractionation methods for in-depth analysis of plasma proteomes. J Proteome Res 11:3090-100
Tang, Hsin-Yao; Beer, Lynn A; Barnhart, Kurt T et al. (2011) Rapid verification of candidate serological biomarkers using gel-based, label-free multiple reaction monitoring. J Proteome Res 10:4005-17
Beer, Lynn A; Tang, Hsin-Yao; Barnhart, Kurt T et al. (2011) Plasma biomarker discovery using 3D protein profiling coupled with label-free quantitation. Methods Mol Biol 728:3-27
Wang, Huan; Tang, Hsin-Yao; Tan, Glenn C et al. (2011) Data analysis strategy for maximizing high-confidence protein identifications in complex proteomes such as human tumor secretomes and human serum. J Proteome Res 10:4993-5005
Tang, Hsin-Yao; Beer, Lynn A; Speicher, David W (2011) In-depth analysis of a plasma or serum proteome using a 4D protein profiling method. Methods Mol Biol 728:47-67