1	Proteomics & Salivary Diagnostics Are We Just Spitting in the Wind?
2	The Reason for the Research THE PUBLIC HEALTH ISSUES
3	THE PUBLIC HEALTH ISSUES No tests for bilateral mastectomies. No tests for BRCA1 & BRCA 2 positive patients. No tests for women under 40. Mammography for women 40-50 have mixed results. Access to care.
4	THE PUBLIC HEALTH ISSUES Compliance Many women obtain breast cancer between yearly mammography visits. Mammography interpretation Legal Risks
5	What is a Biomarker?
6	WHAT SHOULD A TUMOR MARKER BE ABLE TO DO? Marker Usage Screening or early detection. Monitoring the effectiveness of therapy. Detecting tumor recurrence. Differential diagnosis. Prognostic or predictive indicator.
7	Is It Feasible? Is There a Connection Between The Oral Cavity and the Rest of the Body?
8	Histo-Physiologically Similarities Between Tissues
9	Presence of Dental Structures In Remote Tissues
10	Teratoma of the Pituitary Gland
11	SALIVA/BREAST CONNECTION EGF & ITS RECEPTORS Homologous carcinomas of the breast, skin, and salivary glands: A histological immunohistochemical comparison of ductal mammary carcinoma, ductal sweat gland carcinoma, and salivary duct carcinoma.
12	THE CLUE Evidence for the involvement of submandibular gland EGF in mouse mammary tumorgenesis.
13	Why Saliva? Why Not Blood?
14	Why Saliva? Why Not Blood?
15	HAS ANYONE ATTEMPTED TO SEARCH FOR BIOMARKERS IN SALIVA?
16	THE LITERATURE Jenzano JW, et al. J Dent Res 1986: 65(1): 67-70. Jenzano JW, et al. Arch Oral Biol 1988: 33(9): 641-644. Jenzano JW, et al. Arch Oral Biol 1987: 32(10): 757-759. Jenzano JW, et al. Arch Oral Biol1986: 31(9): 627-628.
17	THE LITERATURE Chien DX, et al. Obstet Gynecol 1990 75: 701-4. Navarro MA et al. Breast Can Res Treat 1997: 42: 83-6.
18	Biomarker Development Project Work Plan Hypothesis Discovery phase Find candidate markers Develop classification rules Validation phase Large sample set size Subset of assay conditions Refine classification rules
19	Results
20	Which Breast Malignancy Shall We Target? Invasive Ductal Carcinoma (60% – 80%)* Invasive Lobular Carcinoma Inflammatory Breast Carcinoma Paget’s Carcinoma of the Breast Mucinous, Apocrine, Papillary, Medullary, Cribriform, etc. Turmors of the Male Breast Total: Numerous Pathological types
21	Which Breast Precursor Lesion Shall We Target? Ductal Carcinoma in situ (25% – 30%)* Lobular Carcinoma in situ Microinvasive carcinoma Intraductal Proliferative Lesions Total: 11 Pathological Types of Precursor Lesions
22	Which Benign Breast Shall We Target? Fibroadenomas* Macro-calcifications Micro-calcifications Abcesses Cysts Fibrocystic disease
23	SPECIMEN COLLECTION
24	Why Use Stimulated Whole Saliva? Easy to collect Inexpensive Not subject to circadian rhythm Produces large quantities of saliva Reproducible results
25	SALIVA SPECIMEN STORAGE
26	Salivary Cancer Bank
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29	2D Gel and “Shotgun” Proteome Analyses
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31	Results
32	Protein Overlap
33	Protein Ratios
34	Protein Functions
35	Up Regulated Proteins Less Than p<0.001
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37	STAGING
38	Results We found unique markers to the fibroadenomas The profiling form outside laboratories agreed with our results. Proteins were able to distinguish between node positive and node negative patients. Many of the proteins associated with breast cancer were found in serum, tissues and cell lysates. Same cancer related proteins are present in nipple aspirate fluids. Differences for receptor status.
39	Cancer Genes TB vs. Control
40	Growth & Signaling Pathways
41	Apoptotic Pathway p53
42	POWER BLOTS ANALYSES Over 100 proteins that were over or under expressed. Many were associated with apoptotic and growth factor pathways.
43	OTHER CANCERS
44	HOW DO VERY LARGE, CANCER RELATED PROTEIN ENTER INTO SALIVA?
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48	COLLABORATORS M.D. Anderson Cancer Center Terry Bevers, MD Breast Cancer Oncologist Jennifer Litton, MD Breast Cancer Oncologist Funda Meric Bernstram MD Breast Cancer Oncologist Kevin Coombes, PhD Mathematician & Bioinformatics Adel El-Naggar, Pathologist UT Dental Branch Lenora Bigler PhD Molecular Biology Karen Storthz PhD Molecular Biology William Dubinsky PhD Biochemist & Proteomic Specialist UT Austin John McDevitt PhD Nanotechnology
49	QUESTIONS