LIQUID BIOPSY in tumor diagnosis of cancer

Liquid biopsy non invasive method of tumor diagnosis

1. ROLE OF LIQUID BIOPSY IN TUMOUR DIAGNOSIS PRESENTOR : DR RAHILA MUSTHAFA KC MODERATOR: DR KRISHNARAJ UPADHYAY

2. LIQUID BIOPSY – LATE BLOOMER Liquid biopsy was proposed on 1989 Commercially available : last 6-7yrs Advancement in IT technology – Genomics,PCR,NGS

3. What is a Liquid Biopsy ?? FLUID BIOPSY /FLUID PHASE BIOPSY

4. Liquid biopsy is a minimally invasive test for assessing cancer genetic status, that is present in the plasma component of blood .

5. Why Liquid Biopsies ??

6. FEATURES OF LIQUID BIOPSY

7. DISADVANTAGES OF LIQUID BIOPSY Detection of genetic component is difficult Quantity can be very less (0.01%) Availability of tumour components depends on various factors. Cancer stage,tumor burden etc Emotional stress detecting a cancer that cannot be located or may be slow growing .

8. IS LIQUID BIOPSY SPELLING THE END OF DAYS FOR TRADITIONAL BIOPSY?

9. Formalin-fixation and paraffin-embedding can alter the DNA

10. WHAT TO LOOK FOR IN LIQUID BIOPSY?

11. CIRCULATING TUMOUR CELLS (CTC)

12. Short half life. Low in number - early stage tumors . No definite morphology –varies with cancer type & stage Cluster with leucocytes , fibroblasts,platelets Circulating tumour cells (CTC)-1869

13. FREQUENCY OF CTC

14. TYPES OF CTC Traditional CTC : Proper cancer morphology Express cytokeratin epithelial origin Absence of CD45 ,not of hematopoietic origin Cytokeratin negative CTC ( cancer stem cells which undergoing EMT- most resistant and most prone to metastasis.

15. TYPES OF CTC Apoptotic CTC Traditional CTC Undergoing apoptosis Small CTC CK positive &CD45 negative Size & shape similar to lymphocytes CTC clusters

16. CIRCULATING TUMOR DNA ( CtDNA )

17. HOW TO DIFFERENTIATE NORMAL CIRCULATING DNA VS CtDNA ?

18. Tumor DNA source - majorly from apoptotic and necrotic tumor cells. Under normal physiologic circumstances (10ng-100ng/ml), apoptotic and necrotic remains are cleared by infiltrating phagocytes (Half life-16min to 2.5h). Circulating tumour DNA ( ctDNA )-1948

19. FREQUENCY OF CtDNA

20. SAMPLES FOR EXOSOMES DETECTION BLOOD (PLASMA >SERUM) SALIVA

21. EXOSOMES &MICROVESICLE

22. EXOSOMES OF TUMOUR CELLS

23. SAMPLES FOR EXOSOMES DETECTION BLOOD URINE SALIVA

24. FREQUENCY OF EXOSOMES

25. TECHNIQUES TO ISOLATE & DETECT TUMOR BIOMARKERS

26. CATEGORIZATION OF LIQUID BIOPSY

27. CATEGORIZATION OF LIQUID BIOPSY MATERIAL TYPE ANALYSIS TYPE Analysis of CTC Analysis of CtDNA Analysis of exosomal DNA Analysis of miRNA Mutation analysis EGFR BRAF KRAS KIT Structural change analysis BCR-ABL

28. TECHNIQUES TO ISOLATE CTC

29. FDA APPROVED- breast,prostate ,CRC

30. CLINICAL IMPLICATIONS OF CTC

31. TECHNIQUES TO ISOLATE AND DETECT CtDNA

32. Circulating Tumor DNA ( ctDNA )

33. CLINICAL IMPLICATIONS OF CtDNA

34. ISOLATION & DETECTION OF EXOSOMES

35. EXOSOMES Harbor RNA with tumor -specific mutations Detect gene amplifications and mutations. Potential to detect very early cancer stages

36. CLINICAL IMPLICATION OF EXOSOMES

37. WHO IS BETTER AMONG 3 BIOMARKERS?

38. FUTURE PROSPECTS OF LIQUID BIOPSY

39. PLATELET LIQUID BIOPSY TUMOUR EDUCATED PLATELETS-TEPS

40. TEPS : Platelet RNA undergo changes when in contact with tumour cells. Platelet RNA profile is affected

41. ADVANTAGES OF TEPS Easy to concentrate Second most common cells after Rbcs . Small quantity of blood is needed( fringer prick-a drop of blood)- sufficient to extract mRNA in TEPS. 95% sensitivity and specificity

42. BIOBANKING PRESERVATION Preservation of biologic materials is currently possible CtDNA and Exosomes Large scale trials Shortly provide evidence based elements

43. CURRENT &FUTURE CHALLENGES OF LIQUID BIOPSY

44. TO SUMMARISE;;;;;;;;;;

45. LIQUID BIOPSY PROMISING TECHNIQUE FOR FUTURE PRECISION MEDICINE

46. REFERENCES Raffaele Palmirotta . Liquid biopsy of cancer: a multimodal diagnostic tool in clinical oncology. Ther Adv Med Oncol 2018, Vol. 10: 1–24 . Savonarola A, Palmirotta R, Guadagni F and Silvestris F. Pharmacogenetics and pharmacogenomics : role of mutational analysis in anti-cancer targeted therapy. Pharmacogenomics J 2012; 12: 277–286. Wang J, Chang S, Li G and Sun Y. Application of liquid biopsy in precision medicine: opportunities and challenges. Front Med 2017; 11: 522–527.

47. Perakis S and Speicher MR. Emerging concepts in liquid biopsies. BMC Med 2017; 15: 75. Siravegna G, Marsoni S, Siena S and Bardelli A. Integrating liquid biopsies into the management of cancer. Nat Rev Clin Oncol 2017; 14: 531–548. Crowley E, Di Nicolantonio F, Loupakis F and Bardelli A. Liquid biopsy: monitoring cancergenetics in the blood. Nat Rev Clin Oncol 2013; 10: 472–484. Buder A, Tomuta C and Filipits M. The potential of liquid biopsies. Curr Opin Oncol 2016; 28: 130–134. Zhang W, Xia W, Lv Z, Ni C, Xin Y and Yang L. Liquid Biopsy for Cancer: Circulating Tumor Cells, Circulating Free DNA or Exosomes ? Cell Physiol Biochem2017; 41: 755–768. Ashworth T. A case of cancer in which cells similar to those in the tumors were seen in the blood after death. Med J Aust 1869; 14: 146–147.