Myelodysplasia Acute Myeloid Leukemia Chronic Myelogenous Leukemia Non Hodgkin Lymphoma Chronic Lymphocytic Leukemia Plasma Cell (Multiple) Myeloma
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1 Myelodysplasia Acute Myeloid Leukemia Chronic Myelogenous Leukemia Non Hodgkin Lymphoma Chronic Lymphocytic Leukemia Plasma Cell (Multiple) Myeloma Hodgkin Lymphoma
2 Overview Case Pathophysiology Diagnosis Prognosis Epidemology/Statistics Clinical Treatment
3 Pathology Clinical Patrick J. Buckley, MD, PhD Professor of Pathology Duke University Medical Center Louis F. Diehl, MD Professor of Medicine Duke University Medical Center
4 Hematopoietic Neoplasms Introduction
5 Normal Hematopoiesis
6 Leukemia Monoclonal neoplasm of bone marrow cells typically characterized by a proliferation of immature cells (blasts) in bone marrow/blood Major types are lymphoid and nonlymphoid (the latter commonly called myeloid leukemia) Divided into acute (develops over weeks to months) and chronic (develops over months to years)
7 Lymphoma Monoclonal neoplasm of lymphocytes These lymphocyte proliferations usually form tumors (-omas) in lymph nodes and/or in extranodal lymphoid tissues Major categories are T-cell lymphomas and B-cell lymphomas
8 Tools
9 Tools of the Trade Techniques used for the diagnosis of hematopoietic and lymphoid disorders
10 Morphology
11 Flow Cytometry Fluorochrome-labeled antibodies to cell surface antigens activated by lasers
12 Flow Cytometry Example: Acute Leukemia Normal Marrow Acute Leukemia Blast Region
13 Immunohistochemistry B-cell antigens Antibodies to: T-cell antigens Nuclear antigens CD20 Cyclin D1 CD5
14 Cytochemistry Detection of substances (usually enzymes) associated with certain cell types Used to determine cell lineage in acute leukemia (myeloperoxidase/monocyte esterase) but sometimes for other purposes e.g., tartrate resistent acid phosphatase in hairy cell leukemia
15 Cytochemistry Monocyte esterase in acute monocytic leukemia Myeloperoxidase in AML
16 Cytogenetics Standard karyotype t(8;21) 21q22 Cytogenetics and molecular genetics: some chromosomal changes detected by standard cytogenetics (karyotype analysis) and FISH are of diagnostic and prognostic importance 8q22 FISH
17 Molecular Techniques B- and T-cell clonality studies by pcr to help diagnose lymphoma Detecting translocations (e.g., bcr-abl in CML) and mutations by pcr Sequencing genes (e.g., IgH in CLL) Gene expression studies.and more to come
18 Molecular Techniques Clonality For example, determining that all the B cells in a tissue have the same IgH gene rearrangement allows a diagnosis of lymphoma (as opposed to a polyclonal = reactive process) Rearranged IgH Gene (From Michael Datto, MD, PhD, with permission) V H V H D H J H J H C u C Molecular tests for clonality Capillary electrophoresis of pcr products
19 Disease 1 Myelodysplasia
20 Case 1 Patient 1: 68 year old man seen for pancytopenia picked up during an annual physical examination. One year ago his WBC 4,100, Hct 40.4% and platelet count 145,000. His MCV was 101. He is well. Physical examination: LN: not enlarged ABD: no organomegaly LAB: Hct: 38.2% WBC: 3985 Platelet: 104,000 MCV: 102.3
21 Case 1 Patient 1: 68 year old man seen for pancytopenia picked up during an annual physical examination. One year ago his WBC 4,100, Hct 40.4% and platelet count 145,000. His MCV was 101. He is well. Physical examination: LN: not enlarged ABD: no organomegaly LAB: Hct: 38.2% WBC: 3985 Platelet: 104,000 MCV: 102.3
22 Case 1 History: 68 year old man seen for pancytopenia picked up during an annual physical examination. One year ago his WBC 4,100, Hct 40.4% and platelet count 145,000. His MCV was 101. He is well. Five years ago he had a medical evaluation with WBC 5,300 cells/mm3, Hct 45.5%, platelet count 278,000 cells/mm3 and MCV 92. Physical examination: LN: not enlarged ABD: no organomegaly LAB: Hct: 38.2% WBC: 3985 Platelet: 104,000 MCV: Social History: worked in the manufacture of rubber products from 1953 to 1965.
23 Case 1 Date WBC Hct Platelet MCV 5 years ago , year ago , Now ,
24 Myelodysplasia: Pathophysiology Normal Apoptosis Apoptosis Programmed cell death
25 Myelodysplasia: Pathophysiology Apoptosis Defective: Occurs Early Apoptosis Programmed cell death
26 Myelodysplasia: Pathophysiology Apoptosis Defective: Occurs Early Apoptosis Programmed cell death No Mature Cells
27 Definition - Standard Myelodysplastic syndromes (MDS) are clonal hematopoietic stem cell disorders characterized by ineffective hematopoiesis and peripheral cytopenias. Clonal: All abnormal cells come from a single, common, precursor cell Ineffective hematopoiesis: Immature cells do not develop effective develop into mature cells or the mature cells die early, resulting in too few effective peripheral blood cells
28 Definition - Explanatory The developing hematopoietic cell dies before it reaches maturity.
29 Myelodysplasia: Pathophysiology Genetic Defect #1: Apoptosis Defect Hit #1 Dysplastic Increased Apoptosis Myelodysplasia Normal stem cell Normal cells Anemia Leukopenia Thrombocytopenia Rosenfeld,List. Leukemia 2000 Jan;14(1):2-8
30 Myelodysplasia: Pathophysiology #2: Apoptosis/Maturation Defect Leukemia Hit #1 Hit #2 Dysplastic Increased Apoptosis Myelodysplasia Normal stem cell Normal cells Anemia Leukopenia Thrombocytopenia Rosenfeld,List. Leukemia 2000 Jan;14(1):2-8
31 Myelodysplasia Diagnosis Morphology Cytogenetics
32 Myelodysplasia Diagnosis Pathology
33 Myelodysplasia Morphology
34 Myelodysplastic Syndrome Anemia Thrombocytopenia Hypercellular bone marrow with in increased cell turn-over and death (apoptosis) = ineffective hematopoiesis Leukopenia
35 Dysplastic Erythroid Precursors Normal Erythroid Precursors
36 Dysplastic Erythroid Precursors: Ringed Sideroblasts Abnormal deposition of iron in mitochondria
37 Dysplastic Granulocytes Decreased number of hypogranular, hypolobated granulocytes in blood Normal granulocyte precursors In bone marrow
38 Dysplastic Megakaryocytes Small, dysplastic pawn ball-like megakaryocytes Normal megakaryocytes
39 Myelodysplasia: Diagnosis Cytogenetic Karyotype Normal 53% Abnormal 47% Complex 11% Single/double 36% Single abnormalities Del (5q) 6% -7/del (7q) 4% -7 3% del(7q) 1% +8 6% -Y 2% Sole, Haematologica 90: , 2005
40 Myelodysplasia Classification Disease RA with ringed sideroblasts Refractory anemia (RA) RA with Excess Blasts (RAEB) RA in Transformation (RAEB-T) No affected cell lines Cell ularity No lines with dysplasia Percent blasts Blood Percent blasts BM Median Survival Years 1 N or I 1 < 1% <5% N or I 1 < 1% <5% 4.2 > 2 I 3 < 5% 5-20% 1.5 > 2 I 3 > 5% 21-30%.6
41 Myelodysplasia: Classification Classification and Pathophysiology Leukemia Hit #1 Hit #2 Dysplastic Increased Apoptosis Myelodysplasia Normal stem cell Normal cells Anemia Leukopenia Thrombocytopenia Rosenfeld,List. Leukemia 2000 Jan;14(1):2-8
42 Classification & Prognosis Classification Cell lines affected Blasts in bone marrow Prognosis Blasts Cytogenetics Cytopenias
43 Myelodysplasia:Prognosis Overall Surivival Blood, Vol. 89 No. 6 (March 15), 1997: pp
44 Myelodysplasia:Prognosis Overall Surivival RARS Median Survival: 4 years 10 Year Survival: 30% Blood, Vol. 89 No. 6 (March 15), 1997: pp
45 Myelodysplasia: Statistics Incidence 1/10,000 people per year 7% patients treated with alkylator chemotherapy incidence peaks at 7 years Increasing
46 Myelodysplasia: Statistics Epidemology: Environmental Agriculture chemicals Radiation Solvent Family history Smoking Strom, Leukemia 19:1912, 2005
47 Myelodysplasia: Statistics Epidemology: Chemotherapy Alkylator therapy Topoisomerease inhibitors
48 Myelodysplasia Clinical Features Characteristic Value Age 69+/- 10 Sex (M/F) 1/1 Disease duration (mo) 9.3 +/- 9.8 Hb (g/dl) / ANC x 10(9)/L 2.2 +/-1.8 Platelet count x 10(9)/L 213 +/- 168 Hellstrom-Lindberg, Blood 92 (July) 1998:68-75.
49 Myelodysplasia: Treatment Supportive care Growth factors Transfusions Red blood cells platelets Infection treatment Drug therapy
50 Myelodysplasia: Treatment Azacitadine: Overall Survival Silverman (CALGB) J Clin Oncol 20: , 2002
51 Myelodysplasia: Treatment Azacitadine: Overall Survival Silverman (CALGB) J Clin Oncol 20: , 2002
52 MDS Treated with Azacitidine Quality of Life # Fatigue Dyspnea Social Function Overall QOL Azacitidine Observation Silverman (CALGB) Proceedings ASCO, 1998, Abstract #53 Silverman (CALGB) J Clin Oncol 20: , 2002
53 Myelodysplasia: Treatment Lenalidomide (Revlimid) 43 patients Anemia Transfusion dependent Symptomatic Erythropoietin resistant or not eligible Lenalidomide dosing 25 mg daily PO 10 mg daily PO 10 mg daily for 21/28 days PO Response 24 (56%) responded 20 independence from transfusion 1 Hb increase of 2 g 3 had 50% decrease in transfusion Interstitial deletion involving chromosome 5q % response Adverse events Neutropenia 65% Thrombocytopenia 74% List, N Engl J Med 352: , 2005
54 Myelodysplasia: Treatment Lenalidomide (Revlimid) List, N Engl J Med 352: , 2005
55 Myelodysplasia: Treatment Lenalidomide (Revlimid) List, N Engl J Med 352: , 2005
56 Myelodysplasia: Treatment Lenalidomide (Revlimid) List, N Engl J Med 352: , 2005
57 Myelodysplasia: Treatment Stem Cell Transplant Treatment related mortality: 33% Disease related mortality: 33% Event free survival: 33% Blood, Vol. 95 No. 4 (February 15), 2000: pp
58 Myelodysplasia Summary Developing hematopoietic cells die early No mature cells Slowly the normal cells go away Tendency toward leukemia Survival Long if early and favorable Short if late and unfavorable
59 Disease 2 Acute Myeloid Leukemia
60 Case 2 64 year old man, previous healthy, referred for pancytopenia. He notes some fatigue, progressive dyspnea and easy bruising on the trunk as well as the back and legs. PMH: non contributory PE: normal except for multiple ecchymosis LAB: WBC 2,300 Neut 32% Lymph 45% Mono 15% Atypical lymphs 8% Hct 25% Platelet 45,000
61 Case 2 64 year old man, previous healthy, referred for pancytopenia. He notes some fatigue, progressive dyspnea and easy bruising on the trunk as well as the back and legs. PMH: non contributory PE: normal except for multiple ecchymosis LAB: WBC 2,300 Neut 32% Lymph 45% Mono 15% Atypical lymphs 8% Hct 25% Platelet 45,000
62 Myeloid Cell Development
63 Acute Myeloid Leukemia: Pathophysiology Proceed Through Development Cycle
64 Acute Myeloid Leukemia: Pathophysiology Feedback Control
65 Acute Myeloid Leukemia: Pathophysiology Proceed Through Development Cycle Broken X X
66 Acute Myeloid Leukemia: Pathophysiology Proceed Through Development Cycle Defective X
67 Definition - Explanatory Overproduction of an immature cell that does not develop into a normal mature cell
68 Acute Myelogenous Leukemia: Diagnosis Diagnosis Morphology Immunophenotype Cytogenetics
69 Acute Myelogenous Leukemia Pathology
70 Acute Myeloid Leukemia (AML) Classification and Morphology
71 AML The World Health Organization (WHO) classifies AML according to the presence of certain recurrent genetic abnormalities and whether they arose in the setting of MDS or after chemotherapy All other AML is classified by morphology and stage of differentiation based on the French- American-British (FAB) system
72 AML Without Maturation (for comparison with other subtypes) (FAB AML M1)
73 M1 Bone Marrow Biopsy: Increased Blasts
74 M1 Myeloperoxidase Stain M1 Blood (Blasts)
75 Blast region AML M1: Flow Cytometry
76 AML with Recurrent Genetic Abnormalities
77 AML with t(8;21)(q22;q22) (Core binding factor alpha/eto) 10% of AML with Maturation (FAB AML M2)
78 M2: Bone Marrow Biopsy
79 M2 Bone Marrow Aspirate
80 AML M2: Flow Cytometry Blasts and more mature granulocytes appear to merge CD19+/CD56+ phenotype suggests t(8;21)
81 AML with inv(16)(p13.q22) or t(16;16)(p13.1;q22) (Core binding factor beta/myh11) Acute Myelomonocytic Leukemia with Eosinophils (FAB AML M4 eos )
82 M4 eos Bone Marrow Biopsy
83 Eosinophil Monocyte differentiation Abnormal granules M4 eos : Bone marrow aspirate Blasts Eosinophil
84 AML with t(15;17)(q22;q12) (PML/RARA) Acute Promyelocytic Leukemia (FAB AML M3)
85 APL: Bone Marrow Biopsy (All Promyelocytes)
86 Normal Promyelocytes APL Blood (Abnormal Promyelocytes) APL: Bone Marrow
87 Blast region empty APL: Flow Cytometry
88 Acute Monocytic Leukemia (associated with translocations of 11q23 in some cases, especially children) (FAB AML M5)
89 AML M5: Bone Marrow Biopsy
90 Bone Marrow AM M5 Blood AML M5
91 AML M5 Cytochemical Stain: Monocyte Esterase (ANAE)
92 AML M5: Flow Cytometry Blast area Monocyte area CD4 and CD14 identify monocytes
93 Acute Myelogenous Leukemia: Diagnosis Cytogenetics t(8;21) Normal Chromosomes An exchange of a piece of the long arm of chromosome 8 (ETO gene at 8q22) and the long arm of chromosome 21 (AML1 gene at 21q22). The critical product resulting is bringing the AML1 gene over to chromosome 8. This deregulates the gene product which functions in controlling maturation. Hence, the cells do not mature.
94 Acute Myelogenous Leukemia: Diagnosis Cytogenetics Abnormaity Frequency CR Deaths in Remission Relapse 5 year OS 5 year T(15;17) 12 87% 13% 37% 63% T(8;21) 7 98% 15% 29% 69% Inv(16) 3 88% 9% 42% 61% No abn 42 88% 15% 53% 42% % 12% 44% 48% 11q % 9% 46% 45% % 11% 50% 47% Del(7q) 1 75% 19% 59% 23% Del(9q) 1 100% 9% 39% 60% Complex 5 67% 12% 68% 21% % 8% 80% 10% Del(5q) 1 57% 14% 85% 11% % 12% 90% 4% Grimwade, Blood 92:2322, 1998
95 Acute Myelogenous Leukemia: Classification French American British (FAB) Classification FAB Type Name Comment M0 Undifferentiated Primitive cell; no differentiation Some differentiation to granulocytic M1 Myeloblast series Myeloblast with More differentiation to granulocytic M2 maturation series M3 Promyelocytic Differentiation to progranulocyte series M4 Myelomonocytic Myeloid and monocytoid series M5 Monocytic All monocytic series M6 Erythroleukemia Differentiation to erythroid series M7 Megakaryocytic Differentiation to megakaryocytic series
96 Myeloid Cell Development versus Classification FAB Type Name Comment M0 Undifferentiated Primitive cell; no differentiation Some differentiation to granulocytic M1 Myeloblast series Myeloblast with More differentiation to granulocytic M2 maturation series M3 Promyelocytic Differentiation to progranulocyte series M4 Myelomonocytic Myeloid and monocytoid series M5 Monocytic All monocytic series M6 Erythroleukemia Differentiation to erythroid series M7 Megakaryocytic Differentiation to megakaryocytic series
97 Acute Myelogenous Leukemia: Prognosis Cytogenetics Abnormaity Frequency CR Deaths in Remission Relapse 5 year OS 5 year T(15;17) 12 87% 13% 37% 63% T(8;21) 7 98% 15% 29% 69% Inv(16) 3 88% 9% 42% 61% No abn 42 88% 15% 53% 42% % 12% 44% 48% 11q % 9% 46% 45% % 11% 50% 47% Del(7q) 1 75% 19% 59% 23% Del(9q) 1 100% 9% 39% 60% Complex 5 67% 12% 68% 21% % 8% 80% 10% Del(5q) 1 57% 14% 85% 11% % 12% 90% 4% Grimwade, Blood 92:2322, 1998
98 Acute Myelogenous Leukemia: Prognosis Cytogenetics Prognostic category Favorable Intermediate Poor Abnormality t(8;21) inv(16) t(15:17) Normal +8-5, -5q -7 11q23 Complex Survival 60% 40% 10%
99 Acute Myelogenous Leukemia: Prognosis Cytogenetics Grimwade, Blood 92:2322, 1998
100 Acute Myelogenous Leukemia: Statistics Statistics Median age: case per 20,000 people at age 60 1% of cancer deaths 2.5 deaths per year per 100,000 people Slight increase in males Slight increase incidence in Eastern European Jews
101 Acute Myelogenous Leukemia: Statistics Incidence Rates per 100,000/year ALL AML SEER Incidence Rates
102 Acute Myelogenous Leukemia Epidemiology/Etiology Ionizing radiation Chemicals Drugs alkylating agents cyclophosphamide chlorambucil melphalan topoisomerase II inhibitors etoposide Environmental factors Genetic abnormalities Down s Syndrome Bloom s Syndrome Fanconi s anemia Viruses? Immunological deficiency -No
103 Acute Myelogenous Leukemia: ClinicalClinical Features I Cell deficiency RBC - pallor, fatigue, dyspnea Thrombocytopenia - petechiae, hematoma, bleeding Neutropenia - sepsis, cellulitis, pneumonia Leukemia cutis 10-20% of leukemias CNS leptomeningeal involvement headache, mental status cranial nerves Hyperleukocytosis Blasts > 100,000 - obstruction to capillaries and small arteries
104 Acute Myelogenous Leukemia: ClinicalClinical Features II DIC any form of leukemia very common with M3 (acute promyelocytic leukemia) Tumor lysis syndrome Contents of cells K + PO 4 uric acid
105 Acute Myelogenous Leukemia: ClinicalClinical Features III Leukocytosis low po(2) low glucose high potassium
106 Acute Myelogenous Leukemia: Treatment Overview Induction 3 days of anthracycline 7 days cytarabine Consolidation/Maintenance several more cycles of intensive chemotherapy - highly variable
107 Acute Myelogenous Leukemia: Treatment Schema Daunorubicin 45 mg/m2 x 3 days + Cytarabine 200 mg/m2 CIVI x 7 days Day 14 BM < 5% blasts and < 15% cellularity > 5% blasts and/or > 15% cellularity CR Daunorubicin 45 mg/m2 x 2 days + Cytarabine 200 mg/m2 CIVI x 5 days Cytarabine 3000 mg/m2 IV BID days 1,3,5 (3-4 courses) CIVI = continuous intravenous BM = bone marrow biopsy Blasts = leukemic cells Mayer (CALGB) N Engl J Med 331:904, 1994
108 Acute Myelogenous Leukemia: Treatment Overall Survival and Disease Free Survival Moore, J. O. et al. Blood 2005;105:
109 Acute Myelogenous Leukemia Summary Cells do not mature and do not die Older individual (can be young) Fatigue Normal physicial exam Pancytopenia Aggressive chemotherapy 30% survival
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