diagnosis
play

Diagnosis Greg Wolgamot, MD PhD Workup of leukemia: 1.Blood - PowerPoint PPT Presentation

Diagnosis Greg Wolgamot, MD PhD Workup of leukemia: 1.Blood 2.Lymph node 3.Marrow 4.Prognostic indicators Result Name Result Abnl Normal Range Units WBC 14.7 H 4.0-11.0


  1. Diagnosis Greg Wolgamot, MD PhD

  2. Workup of leukemia: 1.Blood 2.Lymph node 3.Marrow 4.Prognostic indicators

  3. Result Name Result Abnl Normal Range Units WBC 14.7 H 4.0-11.0 K/mm3 RBC 4.51 4.31-5.77 M/mm3 HGB 12.9 L 13.2-17.5 g/dL HCT 38.7 L 38.9-49.9 % MCV 85.8 80.0-100.0 fL MCH 28.6 27.8-33.8 pg MCHC 33.3 31.5-36.5 g/dL RDW 14.0 11.5-14.2 % PLATELET COUNT 126 L 150-400 K/mm3 NEUTRO% 23.6 % LYMPH.% 69.8 % MONO% 5.1 % EOS% 0.6 % BASO% 0.4 % IMM GRAN% 0.5 0-0.5 % Includes myelocytes, metamyelocytes and promyelocytes. NEUTRO# 3.5 1.5-8.0 K/mm3 LYMPH# 10.3 H 1.0-3.5 K/mm3 MONO# 0.7 0.2-1.0 K/mm3 EOS# 0.1 0-0.5 K/mm3 BASO# 0.1 0-0.2 K/mm3 Smudge cells present

  4. PATHOLOGIST INTERPRETATION: Lymphocytosis suggestive of chronic lymphocytic leukemia Flow cytometry would be contributory for further workup John W. Hoyt, MD Pathologist

  5. The Power of Flow Cytometry • Single cell analysis • Multiparametric • Rapid • Quantitative • Flexible

  6. Flourochrome-tagged antibodies flourochrome

  7. Flow Cytometry

  8. Flow Cell

  9. Flow Cytometry

  10. Flow Cytometry

  11. CD45/SS Borowitz et al (1993) AJCP 100:534-40. Steltzer et al (1993) Ann NY Acad Sci 667:265-280

  12. Normal Granulocytic Maturation Wood and Borowitz (2006) Henry ’ s Laboratory Methods

  13. Normal Granulocytic Maturation Wood (2004) Methods Cell Biology 75:559-576

  14. Normal B cell Maturation Wood and Borowitz (2006) Henry ’ s Laboratory Medicine

  15. Abnormal population identification • Normal – Antigens expressed in consistent and reproducible patterns with maturation • Neoplastic – Increased or decreased normal antigens – Asynchronous maturational expression – Aberrant antigen expression – Homogeneous expression

  16. Case 1: Reactive

  17. Case 3: CLL

  18. Lymphoma WOLGAMOT WOLGAMOT

  19. Further Workup 1. Cervical lymph node 2. Bone marrow 3. Prognostic indicators

  20. Case 3 Core biopsy of cervical lymph node

  21. Case 3: CLL Mutated CLL = good prognosis Unmutated CLL = poor prognosis

  22. Case 3: CLL FINAL DIAGNOSIS: Cervical lymph nodes, core biopsy: Chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL). Camilla T. Allen, MD Pathologist

  23. Case 2: Mantle cell lymphoma

  24. Bone marrow aspirate

  25. Case 3: CLL

  26. Result Name Result Abnl Normal Range Units WBC 14.7 H 4.0-11.0 K/mm3 RBC 4.51 4.31-5.77 M/mm3 HGB 12.9 L 13.2-17.5 g/dL HCT 38.7 L 38.9-49.9 % MCV 85.8 80.0-100.0 fL MCH 28.6 27.8-33.8 pg MCHC 33.3 31.5-36.5 g/dL RDW 14.0 11.5-14.2 % PLATELET COUNT 126 L 150-400 K/mm3 NEUTRO% 23.6 % LYMPH.% 69.8 % MONO% 5.1 % EOS% 0.6 % BASO% 0.4 % IMM GRAN% 0.5 0-0.5 % Includes myelocytes, metamyelocytes and promyelocytes. NEUTRO# 3.5 1.5-8.0 K/mm3 LYMPH# 10.3 H 1.0-3.5 K/mm3 MONO# 0.7 0.2-1.0 K/mm3 EOS# 0.1 0-0.5 K/mm3 BASO# 0.1 0-0.2 K/mm3 Smudge cells present

  27. Prognostic Indicators CLL Abnormality Prognosis Very High risk High risk low risk Very low risk same as 10 year survival ------> 29% 37% 57% controls del 17p13.1 (p53) DCI FISH poor; chemo resistance; consider BMT X p53 mutation poor; chemo resistance X BIRC3 very poor; chemo resistance, mutually exclusive to p53 X ZAP70 expression poor X CD38 expression poor X NOTCH associated with Richter's X intermediate risk. Bulky nodes, faster growth, unmutated IgH, requires del 11q22.3 DCI FISH alkylating drugs (cytoxan, bendamustine) X SF3B1 poor. Resistance to fludarabine X trisomy 12 DCI FISH good; low risk. If no NOTCH mutation, low risk X, if NOTCH X, if no NOTCH IgH mutation present good (unmutated is poor) X normal cytogenetics good X del 13q14.3/13q34 DCI FISH good, if isolated X 13p if isolated 13p, good; very low risk X, if isolated t(14;19) ? t(2;14) ?

  28. Cytogenetics & Molecular Studies 1. Cytogenetics 2. FISH (flourescence in situ hybridization)

  29. Case 1: Would be normal

  30. Case 2: Mantle cell lymphoma

  31. Case 3: CLL

  32. Prognostic Indicators CLL Abnormality Prognosis Very High risk High risk low risk Very low risk same as 10 year survival ------> 29% 37% 57% controls del 17p13.1 (p53) DCI FISH poor; chemo resistance; consider BMT X p53 mutation poor; chemo resistance X BIRC3 very poor; chemo resistance, mutually exclusive to p53 X ZAP70 expression poor X CD38 expression poor X NOTCH associated with Richter's X intermediate risk. Bulky nodes, faster growth, unmutated IgH, requires del 11q22.3 DCI FISH alkylating drugs (cytoxan, bendamustine) X SF3B1 poor. Resistance to fludarabine X trisomy 12 DCI FISH good; low risk. If no NOTCH mutation, low risk X, if NOTCH X, if no NOTCH IgH mutation present good (unmutated is poor) X normal cytogenetics good X del 13q14.3/13q34 DCI FISH good, if isolated X 13p if isolated 13p, good; very low risk X, if isolated t(14;19) ? t(2;14) ?

  33. In Situ Hybridization (ISH) Key features: 1) Probe a specific sequence of DNA or RNA 2) Visualize ‘ in situ ’ – within the context of tissue 3) Can be performed on interphase cells 4) Can be performed on non-living/fixed cells

  34. In Situ Hybridization (ISH) 3) Detection method Peroxidase or FITC phosphatase (radiolabel) (fluorochrome: FISH) (enzyme: CISH) S35 S35 S35 * * * . . . . . . . . 2) Probe (DNA) A A C C T A G T T C T G T G G C A G T A G T C A A T A G A A A G T A G T . . . . . . . . 1) Target (DNA or RNA)

  35. ISH Detection Methods: Fluorochrome-labeled Chromogenic (FISH) (CISH)

  36. Uses of In Situ Hybridization  Chromosome enumeration  Locus-specific copy number alterations  Translocation detection t(11;14) translocation by single-fusion FISH  Detection of specific transcripts (RNA) Trisomy 21 HER2 amplification CISH for kappa and by FISH by FISH lambda light chain mRNA  Detection of foreign DNA/RNA Detection of EBV RNA by CISH

  37. Case 2: Mantle cell lymphoma

  38. Fusion Probes vs. Break-Apart Probes for Translocation Detection  Target one locus  Target two loci  Useful when there are  Identifies both partners numerous potential involved in a translocation partner genes

  39. Case 2: Mantle cell lymphoma Fusion probes

  40. Case 3: CLL

  41. Prognostic Indicators CLL Abnormality Prognosis Very High risk High risk low risk Very low risk same as 10 year survival ------> 29% 37% 57% controls del 17p13.1 (p53) DCI FISH poor; chemo resistance; consider BMT X p53 mutation poor; chemo resistance X BIRC3 very poor; chemo resistance, mutually exclusive to p53 X ZAP70 expression poor X CD38 expression poor X NOTCH associated with Richter's X intermediate risk. Bulky nodes, faster growth, unmutated IgH, requires del 11q22.3 DCI FISH alkylating drugs (cytoxan, bendamustine) X SF3B1 poor. Resistance to fludarabine X trisomy 12 DCI FISH good; low risk. If no NOTCH mutation, low risk X, if NOTCH X, if no NOTCH IgH mutation present good (unmutated is poor) X normal cytogenetics good X del 13q14.3/13q34 DCI FISH good, if isolated X 13p if isolated 13p, good; very low risk X, if isolated t(14;19) ? t(2;14) ?

  42. Question 2: FISH: 1. Include both osteichthyes and chondrichthyes. 2. Is a molecular technique that can detect specific sequences of DNA. 3. Allows classification of leukemias based on the proteins the cells express.

  43. Lymphoma WOLGAMOT WOLGAMOT

  44. Lymphoma WOLGAMOT WOLGAMOT

  45. Special thanks to: Ryan Fortna , MD PhD NW Pathology Brent Wood, MD PhD U of Washington Cancer Program at SJH

Recommend


More recommend