1. Obtain resected tissue from Breast Cancer Research foundation Obtain 5+ canines (6mos-1yr), mix of Labradors and German shepherds Train canines to indicate for breast cancer VOC’s Identify VOC’s in resected tissue samples Test other types of cancer using chemistry and canines for VOC’s Future research to identify odor signatures of other cancers Detection and Identification of Volatile Organic Compounds in Breast Cancer: Proposal for Research Makensie Kraft Department of Biological Sciences York College of Pennsylvania Introduction 1.Studies have shown that canines are capable of detecting odor signatures specific to a type of cancer in urine 1, breath 2, skin 3, and tissue 4 samples. The odor signatures are composed of volatile organic compounds, mostly benzene and alkane derivatives 5. 2.In 2009, over 200,000 women were diagnosed with breast cancer; over 40,000 women died from breast cancer (CDC). If the odor signature could be identified, it would make progress towards more effective diagnostic methods. 3.Different grades of one cancer type emit the same odor signature 4, detectable by canines, which is independent from other symptoms of infection, such as bleeding, infection, and inflammation 2. 4.In Canada 6, the UK 7, and the US 8, canines are being trained for biomedical purposes. People with specific diseases can adopt medically trained dogs to alert them of abnormalities. Cancer-detection canines are currently used for research and organizations are seeking funds to actively deploy trained canines into hospitals and communities 8. 5.Although previous studies have shown that canines can detect the odor signature, there was little research on the chemistry behind it. Main Objectives 1.Identify the volatile compounds of breast cancer using GC-MS Olfactometry 2.Train canines to detect the identified volatile compounds using sequential phases training methods. 3.Identify an odor signature unique to breast cancer by testing different types and grades of cancerous tissues. Literature Review Canines are capable of detecting multiple odor signatures and will retain this information for long periods of time 9. Canines were trained to detect melanoma in skin samples and were able to identify the exact gauze patch with the cancerous skin underneath, which is evidence for their specificity 3. Canines were trained to detect kidney and bladder cancer in urine samples. The canines could distinguish cancerous samples from others with infection, inflammation, or bleeding 1. Canines were trained to detect different types and grades of ovarian cancer in tissue samples. By using 31 different histopathological types of ovarian cancer, they determined that the common ovarian carcinomas emit the same odor signature, which also differs from cervical, endometrial, and vulvar carcinomas. 4 Volatile organic compounds (VOC) of lung cancer were identified using automated thermal desorption, gas chromatography, and mass spectroscopy 10. Proposed Methods Expected Results 1.Volatile compounds found in cancerous tissues, not in healthy tissues, most likely benzene or alkane derivatives. 2.Canines able to indicate for VOC’s of breast cancer and distinguish them from other odors in a sample. 3.A unique odor signature for breast cancer, found in the most common grades, that differs from other types of cancer. Acknowledgements I would like to thank Dr. Rehnberg and all of the biology faculty that helped evaluate and improve my proposal. I would also like to thank Dr. Fautch for helping my understanding of the chemistry required for this proposal. Literature Cited 1.Willis CM, Church SM, Guest CM, Cook WA, McCarthy Noel, Bransbury AJ, Church MRT and Church JCT. 2004. Olfactory detection of human bladder cancer by dogs: Proof of principle study. British Medical Journal. 329: 712-714 2.McColloch, Michael; Jezierski, Tadeusz et. al. 2006. Diagnostic accuracy of canine scent detection in early and late stage lung and breast cancers. Integrative Cancer Therapies. 5(1) : 30-39. 3.Pickel Duane, Manuncy GP, Walker DB, Hall SB, Walker JC. 2004. Evidence for canine olfactory detection of melanoma. Applied Animal Behaviour Science. 89: 107-116. 4.Horvath et al. 2008. Human Ovarian Carcinomas Detected by Specific Odor. Integrative Cancer Therapies. 7(2): 76-80 5.Shirasu, M and Touhara, K. 2011.The scent of disease: volatile compounds of the human body related to disease and disorder. Journal of Biochemistry. 150(3): 257-266. 6." Trailrunners K9 Training Center and Labrador Retrievers in Canada – Cancer Early Detection." CancerDogs. Trailrunners K9 Training Center, 2011. Web.. 7."Cancer Detection." Medical Detection Dogs. Royal Canin, 2013. Web.. 8."Saving Lives." Dogs Detect Cancer. InSitu, 2012. Web.. 9.Johnston, J. M. 1999. Canine detection capabilities: Operational implications of recent R & D findings. Institute for Biological Detection Systems. Auburn University. 10.Phillips, M. "Detection of Lung Cancer With Volatile Markers in the Breath." Chest 123.6 (2003): 2115-123. Print. 11.Wang, Xiao-Bo et al. 2000. Cell Separation by Dielectrophoretic Field Flow Fractionation. Analytical Chemistry. 72 (4), 832-839 *5 Dielectrophoretic Field-Flow Fractionation (DEP-FFF) has been shown to separate cancerous cells from blood samples as shown here 11. Canines were trained to detect breast and lung cancer in breath samples 2. This study used sequential phase training, shown here, which proved effective. Chemical Analysis GC-MS Olfactometry to identify VOC’s DEP-FFF to isolate cancerous cells GC-MS Olfactometry to compare histopathology Canines Training Phases – positive indication for VOC’s Testing Phases- sensitivity and specificity Repeat testing used to confirm presence or absence of odor signature