1. Saliva Diagnostics and Cancer Centre, JBR Society
Saliva Diagnostics:Cancer
JBR Saliva Diagnostic Technologies: The next frontier in cancer diagnostics?
Balwant Rai
Saliva Diagnostics and Cancer Centre, JBR Society

2. Introduction
Cancer cases are still diagnosed at the advanced stage Survival for certain cancers –improved-quite low New diagnostic tool -potentially improve cancer diagnosis
Wilkinson.2014

3. Introduction
Human saliva-mirrors our body’s health Role- monitoring oral and systemic health Hormones, antimicrobial constituents, proteins, enzymes, antibodies, cytokines, ions and minerals Energetic and future area in diagnosis & prognosis-physio-biochemical alterations
Pfaffe et al. 2011; Rai et al. 2011

4. Saliva
Whole saliva-3 pairs of major salivary glands 60%-submandibular gland 30%-parotid gland 5%-sublingual gland 7%-minor salivary gland
Rai. 2017

5. 0.5-organic and inorganic
Composition and Functions
Anti-bacterial
Viscoelasticity
Buffering
Digestion
Anti-viral
Saliva composition
99.5%-water
0.5-organic and inorganic
pH:
Mineralization
Anti-fungal
Lubrication
Tissue coating
Rai and Kaur 2017

6. Methods of Collection of Saliva
Whole Saliva Catheterization Parotid Saliva Submandibular (Submaxillary) Saliva
Kaur et al. 2011; Rai et al. 2010

7. Standardization of Collection
Resting secretions- at least 5 minutes Stimulated saliva-Consistent gustatory stimulant Flow rate-Timed, focus on specific gland Time of day-Circadian rhythm, time

8. Saliva as a Mirror of the Body
Tissue fluid levels of natural substances Therapeutic Emotional status Hormonal status Immunological status Neurological status Nutritional and metabolic influences
Rai et al. 2009

9. Saliva as a Mirror of the Body
Sjögren’s syndrome- lymphoepithelial lesions Cystic fibrosis Hormonal dysfunction- diabetes, pancreatitis, adrenal-cortex disease, thyroid disease, acromegaly, menopause Hypertension Obesity and hyperlipidemia Alcoholic cirrhosis Malnutrition Neurologic diseases- Parkinson’s disease, Bell’s and cerebral Psychogenic diseases Oral diseases Oncology
Rai et al. 2009; Wong et al.2008

10. Design parameters for monitoring Device
Real-time Automatic Non-invasive Multiple biomarkers Very small samples High sensitivity and specificity Low in mass, small in volume Fluctuations in Different parameters No waste artifacts and noninfectious Heavy data storage capabilities and reusable Not time consuming to use and little or no skill required for use
Rai and Kaur. 2012

11. The panel of 40 Saliva Based Biomarkers for Cancer
JBR SaliCan chipTM
The panel of 40 Saliva Based Biomarkers for Cancer
Rai and Kaur. 2012

12. Patients Selection

13. Rai et al.2007 Salivary vitamins E and C in oral cancer
Lipid peroxidation may be involved in cancer-free radicals Salivary vitamins E and C: 50 oral cancer and 24 healthy persons Significantly lower levels of vitamins E and C were observed in oral cancer patients Antioxidant nutrients may be utilized to a greater extent in oral cancer patients to counteract free radical-mediated cell disturbances
Rai et al.2007

14. Tongue cancer-most common and fatal types of cancers in the world Abnormalities in purine metabolism- many human tumors Fifty patients with squamous cell carcinoma of the tongue and 30 normal subjects Salivary adenosine deaminase (ADA) activity- assessed
Adenosine deaminase in saliva as a diagnostic marker of squamous cell carcinoma of tongue
Rai et al. 2011

15. Statistically significant differences between test and control groups were observed in salivary ADA (P < 0.001). Salivary ADA levels significantly increased as the disease stage progressed from stage I to stage III of squamous cell carcinoma of the tongue in both genders (P < 0.001) Salivary ADA might be used as a diagnostic tool for early detection of squamous cell carcinoma of tongue.
Adenosine deaminase in saliva as a diagnostic marker of squamous cell carcinoma of tongue
Rai et al. 2011

16. Rai and Kaur. 2013 Salivary Biomarkers: Breast cancer patients
Angiogenic and tumor markers in saliva could be complementary to the current methods used for breast cancer diagnosis Materials and Methods: 20 healthy individuals and 30 breast cancer patients
Rai and Kaur. 2013

17. Rai et al.2017 Salivary Biomarkers: Breast cancer patients
The inclusion criteria: Not pregnant or lactating Not any active oro-dental disease Not prior-carcinoma  Diagnosis of breast cancer Control subjects-healthy volunteers Saliva samples
Rai et al.2017

18. Salivary Biomarkers: Breast cancer patients
Salivary biomarkers: Carcinoembryonic antigen (CE) (ng/ml) Vascular endothelial growth factor (VF) (ng/ml ) Epidermal growth factor (EF)  (ng/ml) SaliBrTM
Rai et al.2017

19. Rai et al.2017 Salivary Biomarkers: Breast cancer patients
Salivary biomarkers levels
Rai et al.2017

20. ROC curve analysis on the logistic regression models.
Salivary Biomarkers: Breast cancer patients
ROC curve analysis on the logistic regression models.
AUC (%)
Sensitivity (%)
Specificity (%) CE 62 56 60 VF 45 52 EF 51 43 50
SaliBr 89 81 78
Rai et al.2017

21. Rai et al.2017 Salivary Biomarkers: Breast cancer patients
Saliva is a novel-Breast cancer Combination: Salivary biomarkers marker, standard screening physical exam and mammogram
Rai et al.2017

22. Xiao et al. 2012 Saliva Biomarkers:Lung Cancer Patients
Lung cancer-asymptomatic-early stages Early detection-reduce lung cancer burden Saliva - attractive diagnostic fluid Profiling of salivary biomarkers-disease progression
Saliva Biomarkers:Lung Cancer Patients
Xiao et al. 2012

23. 30 lung cancer samples-diagnosed as lung cancer by using biopsy 30 healthy control samples Saliva samples ELISA
Saliva Biomarkers:Lung Cancer Patients

24. Saliva Biomarkers:Lung Cancer Patients
Demographics
Saliva Biomarkers:Lung Cancer Patients
Cancer
Control
Age (years)
61.3(11.4)
62.04(10.5)
Smoke History 26 25
Lung Cancer 30
Sex (M:F)
14:16
15:15

25. Rai et al. 2017 Saliva Biomarkers:Lung Cancer Patients
HP, ANXA1, AZGP1, Interleukin 1 receptor antagonist, S100 calcium binding protein A8, human cal protectin and SaliLc Salivary biomarkers panel (HP, human calprotectin and SaliLc)
Saliva Biomarkers:Lung Cancer Patients
Rai et al. 2017

26. Performance of different biomarker combination
Saliva Biomarkers:Lung Cancer Patients
Sensitivity%
Specificity%
AUC
SaliLc 82 73
0.92 HP 62 60
0.89
Human calprotectin 78 75
HP, human calprotectin and SaliLc 89 87
0.93
Rai et al. 2017

27. Rai et al. 2017 Saliva Biomarkers:Lung Cancer Patients
Salivary biomarkers-develop lung cancer The discriminatory power-lung cancer clinical screening and detection
Saliva Biomarkers:Lung Cancer Patients
Rai et al. 2017

28. Subbannayya et al. 2015 Saliva Biomarkers: Gastric Cancer Patients
Gastric cancer-third most lethal cancer in the world Endoscopy method-benign and malignant diseases Only few studies on salivary biomarkers
Saliva Biomarkers: Gastric
Cancer Patients
Subbannayya et al. 2015

29. Rai et al. 2015 Saliva Biomarkers: Gastric Cancer Patients
30 Gastric cancer and 20 healthy Saliva sampling Salivary biomarkers and microbiome
Saliva Biomarkers: Gastric
Cancer Patients
Rai et al. 2015

30. Rai et al. 2017 Saliva Biomarkers: Gastric Cancer Patients
miR-10b, miR-144, miR-21 and miR-451 Capnocytophaga gingivalis, Prevotella melaninogenica, Streptococcus mitis, Leptotrichia and Porphyromonas 
Saliva Biomarkers: Gastric
Cancer Patients
Rai et al. 2017

31. Rai et al. 2017 Saliva Biomarkers: Gastric Cancer Patients
Salivary biomarkers, multiple salivary bacteria, metabolite profiles in Gastric Cancer
Saliva Biomarkers: Gastric
Cancer Patients
Rai et al. 2017

32. Xie et al. 2013 Salivary Biomarkers: Pancreatic Cancer Patients
Pancreatic cancer (PC)-fourth-cancer death MicroRNAs-PC Saliva-source of miRNAs Differences in salivary microRNA profiles-precancerous lesions, inflammatory disease and PC
Salivary Biomarkers: Pancreatic Cancer Patients
Xie et al. 2013

33. Rai et al. 2017 Salivary Biomarkers: Pancreatic Cancer Patients
Pancreatic cancer (n = 15), pancreatitis (n =10 ) and healthy controls (n = 10)-endoscopic examination. Saliva Candidate miRNAs -q(RT)PCR
Salivary Biomarkers: Pancreatic Cancer Patients
Rai et al. 2017

34. Rai et al. 2017 Salivary Biomarkers: Pancreatic Cancer Patients
Average Cq values, sensitivity and specificity of the candidate microRNAs
Salivary Biomarkers: Pancreatic Cancer Patients
Conrol
Pancreatitis PC
Spécificity
sensitivity
Let-7c
40.3
29.2
21.4 93 90
hsa-miR-210
36.4
30.2
20.4
100 78
hsa-miR-23b
40.4
34.5
25.3 80
Rai et al. 2017

35. Salivary miRNA -biomarkers for distinguishing patients Let-7c hsa-miR-210 hsa-miR-23b
Salivary Biomarkers: Pancreatic Cancer Patients
Rai et al. 2017

36. Turan et al. 2013 Salivary Biomarkers: Prostate Cancer Patients
Prostate cancer-second leading cause of cancer death in men Histopathological analysis (Gleason score) and serum prostate-specific antigen (PSA) PSA-weaknesses-benign prostatic hyperplasia-aggressive prostate cancer Good candidate salivary biomarkers
Turan et al. 2013

37. Rai et al. 2015 Salivary Biomarkers: Prostate Cancer Patients
PC (Prostate Cancer) patients (n = 23) and age-matched healthy controls (n = 23) Saliva sampling Fast ultrahigh performance liquid chromatography-tandem mass spectrometry  Taurocholic acid,Glycocholic acid, 5-hydroxy-L-tryptophan,and chenodeoxycholic acid
Rai et al. 2015

38. Rai et al. 2015 Salivary Biomarkers: Prostate Cancer Patients
PC (Prostate Cancer) patients (n = 23) and age-matched healthy controls (n = 23) Saliva sampling Fast ultrahigh performance liquid chromatography-tandem mass spectrometry  Taurocholic acid,Glycocholic acid, 5-hydroxy-L-tryptophan,chenodeoxycholic acid and SaliPr
Rai et al. 2015

39. Turan et al. 2013 Salivary Biomarkers: Prostate Cancer Patients
ROC analysis revealed to be taurocholic acid,Glycocholic acid, 5-hydroxy-L-tryptophan,chenodeoxycholic acid and SaliPr potent discriminators between PC and control groups. Metabolomics platform could improve the diagnosis of diseases.
Turan et al. 2013

40. American Cancer Society. 2016
Salivary Biomarkers: Ovarian Cancer Patients
Ovarian cancer-lethal gynaecological malignancy Five- and 10-year-survival rates of 45% and 35%, respectively Diagnosed at the advanced stages- massive peritoneal metastasis
American Cancer Society. 2016

41. Rai et al. 2017 Salivary Biomarkers: Ovarian Cancer Patients
15 normal healthy women, 20 benign pelvic masses, and 20 malignant pelvic tumors Saliva sampling ELISA
Rai et al. 2017

42. Rai et al. 2017 Salivary Biomarkers: Ovarian Cancer Patients
Saliva CA 125 value greater than 3000 U/mL as the positivity criteria SaliOv value greater than 40 U/mL as the positivity criteria Sn of the saliva CA 125 and SaliOv were 67 and 90 respectively Sn of the saliva CA 125 and SaliOv were 65 and 89 respectively
Rai et al. 2017

43. Rai et al. 2017 Salivary Biomarkers: Ovarian Cancer Patients
Saliva CA 125 and SaliOv levels may be a new way of screening for malignant ovarian tumors
Rai et al. 2017

44. Saliva Diagnostics Powered NanoTechnologies Proteomics Genomics
Salivary Biomarkers: Ovarian Cancer Patients

45. Saliva
Diagnostics Pharmaco-proteomics Pharmaco-genomics Personalized Individual Medicine
Rai et al. 2017

46. Rai et al. 2017 Future Perspectives
A major limitation in saliva research-specific disease biomarkers-high sensitivity and specificity Standardization No validation Multicentre studies with larger sample sizes
Rai et al. 2017

47. Point of Care

48. Rai B . 2017 Fellowship of Saliva Diagnostics
Fellowship of Saliva Diagnostics: 100 hours Certification in Saliva Diagnostics: 40 hours
Rai B

49. Conclusion
Sialochemistry provides important information Greater acceptance Non-invasive May open door to new
Rai B.2017

50. Acknowledgement
JBR society Saliva diagnostics Group BNS, Punjab World dental Network Open Research Hub Incudent
Rai B.2017

51. Thanks