Salivary Biomarkers for Early Detection of Breast Cancer

Institution: University of California, Los Angeles
Investigator(s): Lei  Zhang , Ph.D. -
Award Cycle: 2010 (Cycle 16) Grant #: 16IB-0004 Award: $150,000
Award Type: IDEA
Research Priorities
Innovative Treatments>Biomarkers and novel screening approaches: unmasking the hidden signs



Initial Award Abstract (2010)

This is a translational research project towards a non-invasive early detection of breast cancer. The high burden of breast cancer in women worldwide underscores the unmet potential of biomarker for early detection. Early detection is the key to positive, long-lasting outcomes, thus reducing the suffering and cost to society associated with the disease. There is a pressing need for novel strategies and new sources for discovering breast cancer biomarkers. A sensitive assay to identify biomarkers that can accurately diagnose the onset of breast cancer using non-invasively collected clinical specimens is ideal for early detection.

Our long-term goal is to develop a novel early detection tool for breast cancer. We hypothesize that there are clinically discriminatory biomarkers in saliva for the early detection of breast cancer. Using a developed salivary transcriptomic approach, we have discovered and pre-validated eight mRNA biomarkers in saliva for detecting breast cancer. The objective of this proposal, which is the essential step toward attaining our long-range goal, is to evaluate the discovered salivary mRNA biomarkers for detecting early breast cancer by focusing on pre-invasive lesions (stage 0, non-invasive breast cancers, such as DCIS.) and early stage (stage I and IIA) breast cancer, in a new patient population. The study design will adhere to the PRoBE principle (prospective specimen collection and retrospective blinded evaluation) set forth recently by the NCI Early Detection Research Network (EDRN). Our strategy for this study is: 1) use PRoBE design to collect new samples that includes DCIS, Stage I and IIA IDC; and 2) evaluate current biomarkers by RT-qPCR using the new cohort. A “Go or No-go” decision will be made by evaluating the biomarkers’ performance.

The outcome of this project will validate salivary transcriptomic biomarkers for the non-invasive early detection of breast cancer, leading to a single institution validation, followed by PRoBE-designed multi-institution validation and FDA level pivotal clinical validation. We are optimistic that the salivary transcriptomic biomarkers will meet the rigor of these evaluations and emerge as highly specific and sensitive tools for the early detection of breast cancer in saliva, a non-invasive biofluid.




Final Report (2012)

The objective of this project was to evaluate the performance and translational utilities of the recently discovered salivary transcriptomic biomarkers for detecting early stage breast cancer by focusing on pre-invasive lesions (stage 0, non-invasive breast cancers, such as DCIS) and early stage (stage I and IIA) breast cancer, in a new patient cohort.

We first discovered seven new gene biomarker candidates using systems biological approaches. Together with eight gene biomarkers discovered previously (Zhang et aI., 2010), these biomarker candidates were tested using 57 new cases and 67 healthy controls using R T -qPCR. Seven genes were validated, including CSTA, IGF2BP1, MDM4, S100A8, CDC14B, PDCD1LG2, and MBD1, which yield ROC-plot AUC values between 0.57 and 0.75. Using logistic regression, the accuracy, sensitivity and specificity of 4-validated-biomarker combination (CSTA, MDM4, CDC14B, and PDCD1LG2) on the new sample set (n =124) were 82%, 92.9% and 78.6%, respectively. The validated salivary rnRNA biomarkers possess the discriminatory power for the early detection of breast cancer, poising them for the initiation of a multi-center validation in a definitive clinical context.

Additionally, we added a new project aims to explore the potential of next-generation sequencing (NGS) technologies for new biomarker discovery in saliva samples. Recent development of NGS technology has enabled us to sequence dozens of genes in various patient samples using a simple workflow. We tested the Ion PGM platform at the UCLA Clinical Microarray Core, of which the gene expression levels can be examined with short turnaround time and low cost, starting with a single-tube reaction and 10 ng of RNA. This could be an ideal solution and hold a great potential• for diagnostic applications. In principle, this NGS-based test could potentially be developed for any cancer patient, and used not only for early detection of recurrent cancer in the same patient in the future, but also to monitor an individual's response to different therapeutic options. This method represents an important foray into offering truly personalized cancer diagnostics.




Symposium Abstract (2010)

Lei Zhang, Hua Xiao, Scott Karlan, Michael Zhou, Jenny Gross, David Elashoff, David Akin, Xinmin Yan, David Chia, Beth Karlan, and David T. Wong

1 School of Dentistry and Dental Research Institute, University of California-Los Angeles, Los Angeles, California
2 Cedars-Sinai Medical Center, Women's Cancer Research Institute, Los Angeles, California, 90048-1865, USA
3 Saul & Joyce Brandman Breast Center, Cedars-Sinai Medical Center, Los Angeles, California, 90048-1865, USA
4 Department of Biostatistics, School of Public Health, University of California-Los Angeles, Los Angeles, California
5 Jonsson Comprehensive Cancer Center, University of California-Los Angeles, Los Angeles, California
6 Division of Hematology & Oncology, David Geffen School of medicine, University of California-Los Angeles, Los Angeles, California
7 Division of Head and Neck Surgery/Otolaryngology, David Geffen School of Medicine, University of California-Los Angeles, Los Angeles, California
8 Department of Pathology and Laboratory Medicine, Jonsson Comprehensive Cancer Center, University of California-Los Angeles, Los Angeles, California
9 Henry Samueli School of Engineering and Applied Science, University of California-Los Angeles, Los Angeles, California

A sensitive assay to identify biomarkers using non-invasively collected clinical specimens is ideal for breast cancer detection. In this study, a case-control discovery and independent biomarker pre-validations were conducted to evaluate the performance and translational utilities of salivary transcriptomic and proteomic biomarkers for breast cancer detection. Salivary transcriptomes and proteomes of 10 breast cancer patients and 10 matched controls were profiled using Affymetrix HG-U133-Plus-2.0 Array and two-dimensional difference gel electrophoresis (2D-DIGE), respectively. Preclinical validations were performed to evaluate the discovered biomarkers in an independent sample cohort of 30 breast cancer patients and 63 controls using RT-qPCR (transcriptomic biomarkers) and quantitative protein immunoblot (proteomic biomarkers). Transcriptomic and proteomic profiling revealed significant variations in salivary molecular biomarkers between breast cancer patients and matched controls. Eight mRNA biomarkers and one protein biomarker, which were not affected by the confounding factors, were pre-validated, yielding an accuracy of 92% (83% sensitive, 97% specific) on the preclinical validation sample set. These findings support that transcriptomic and proteomic signatures in saliva can serve as biomarkers for the non-invasive detection of breast cancer.



Discovery and preclinical validation of salivary transcriptomic and proteomic biomarkers for the non-invasive detection of breast cancer.
Periodical:PLOS One
Index Medicus: PLOS One
Authors: Zhang L, Xiao H, Karlan S, Zhou H, and Wong D
Yr: 2010 Vol: 5 Nbr: 12 Abs: Pg:e15573