3/21/16 Objecves Gene+cs of Select • Define tumor suppressor gene and oncogene Lymphoproliferave Neoplasms: • Correlate genotypes Pathophysiology, Prognos+caon, and Therapy with prognosis in lymphoproliferave neoplasms Kris+n Landis-Piwowar PhD, MLS(ASCP)CM • Recognize the targets of lymphoproliferave Clinical Laboratory Hematology, 3/E, McKenzie • Williams neoplasm therapies 2 Leukemia Lymphoma WHO 2008: Mature B-Cell Neoplasms • Chronic lymphocy+c leukemia/small lymphocy+c • Mantle cell lymphoma lymphoma • Diffuse large B-cell lymphoma (DLBCL), NOS • Primarily BM and • Primarily lymph • B-cell prolymphocyc leukemia • T-cell/his+ocyte rich large B-cell lymphoma • Primary DLBCL of the CNS • Splenic marginal zone lymphoma • Primary cutaneous DLBCL, leg type blood involvement node / solid ssue • Hairy cell leukemia • EBV-posi+ve DLBCL of the elderly • Splenic lymphoma/leukemia, unclassifiable • DLBCL associated with chronic inflammaon • Splenic diffuse red pulp small B-cell lymphoma • Lymphomatoid granulomatosis • Hairy cell leukemia variant involvement • Primary medias+nal (thymic) large B-cell lymphoma • Myeloid or • Lymphoplasmacy+c lymphoma • Intravascular large B-cell lymphoma • Waldenström macroglobulinemia • ALK-posi+ve large B-cell lymphoma lymphoid origin • Lymphoid origin • Heavy chain diseases • α Heavy chain disease • Plasmablas+c lymphoma • γ Heavy chain disease • Large B-cell lymphoma arising in HHV8-associated • May secondarily • May secondarily • μ Heavy chain disease mul+centric Castleman disease • Plasma cell myeloma • Primary effusion lymphoma • Solitary plasmacytoma of bone • Burkih lymphoma involve lymph involve BM & • Extraosseous plasmacytoma • B-cell lymphoma, unclassifiable, with features intermediate • Extranodal marginal zone lymphoma of mucosa-associated between diffuse large B-cell lymphoma and Burkih nodes and solid blood; “leukemic lymphoid +ssue (MALT lymphoma) lymphoma • Nodal marginal zone lymphoma • B-cell lymphoma, unclassifiable, with features intermediate ssues phase” • Pediatric nodal marginal zone lymphoma between diffuse large B-cell lymphoma and classical • Follicular lymphoma Hodgkin lymphoma • Pediatric follicular lymphoma • Primary cutaneous follicle centre lymphoma Adapted from: Clinical Laboratory Hematology, 3/E, McKenzie • Williams 4 3 WHO 2008: Mature T-Cell Neoplasms WHO 2008: More Neoplasms • T-cell prolymphocyc leukemia • • Hodgkin lymphoma • B lymphoblas+c leukemia/lymphoma Sézary syndrome • Nodular lymphocyte predominant Hodgkin lymphoma • B lymphoblas+c leukemia/lymphoma, NOS • T-cell large granular lymphocy+c • Primary cutaneous CD30+ T-cell • Classical Hodgkin lymphoma • B lymphoblas+c leukemia/lymphoma with recurrent leukemia • Nodular sclerosis classical Hodgkin lymphoma gene+c abnormali+es lymphoproliferave disorders • Lymphocyte-rich classical Hodgkin lymphoma • B lymphoblas+c leukemia/lymphoma with t(9;22) (q34;q11.2);BCR-ABL 1 • • Chronic lymphoproliferave disorder of • Lymphomatoid papulosis Mixed cellularity classical Hodgkin lymphoma • B lymphoblas+c leukemia/lymphoma with t(v; NK cells • Lymphocyte-depleted classical Hodgkin lymphoma 11q23);MLL rearranged • Primary cutaneous anaplas+c large cell • B lymphoblas+c leukemia/lymphoma with t(12;21) • Aggressive NK-cell leukemia lymphoma (p13;q22) TEL-AML1 (ETV6-RUNX1) • His+ocy+c and dendri+c cell neoplasms • B lymphoblas+c leukemia/lymphoma with • Systemic EBV-posi+ve T-cell • Primary cutaneous γδ T-cell lymphoma • His+ocy+c sarcoma hyperdiploidy lymphoproliferave disease of • Langerhans cell his+ocytosis • B lymphoblas+c leukemia/lymphoma with hypodiploidy • Langerhans cell sarcoma • B lymphoblas+c leukemia/lymphoma with t(5;14) • Primary cutaneous CD8+ aggressive (q31;q32) IL3-IGH childhood • Interdigitang dendri+c cell sarcoma epidermotropic cytotoxic T-cell • B lymphoblas+c leukemia/lymphoma with t(1;19) • Hydroa vacciniforme-like lymphoma • Follicular dendri+c cell sarcoma (q23;p13.3);TCF3-PBX1 lymphoma • Fibroblas+c re+cular cell tumor • Adult T-cell leukemia/lymphoma • Intermediate dendri+c cell tumor • T lymphoblas+c leukemia/lymphoma • Primary cutaneous CD4+ small/medium • • Extranodal NK/T-cell lymphoma, nasal Disseminated juvenile xanthogranuloma T-cell lymphoma • Acute leukemias of ambiguous lineage type • Poshransplantaon lymphoproliferave disorders • Peripheral T-cell lymphoma, NOS (PTLDs) • Acute undifferen+ated leukemia • Enteropathy-associated T-cell • Mixed phenotype acute leukemia with t(9;22)(q34;q11.2); • Early lesions BCR-ABL1 lymphoma • Angioimmunoblas+c T-cell lymphoma • Plasmacy+c hyperplasia • Mixed phenotype acute leukemia with t(v;11q23); MLL • Hepatosplenic T-cell lymphoma • Infec+ous mononucleosis–like PTLD rearranged • Anaplas+c large cell lymphoma, ALK- • Polymorphic PTLD • Mixed phenotype acute leukemia, B-myeloid, NOS • • Subcutaneous panniculi+s-like T-cell posive Monomorphic PTLD (B- and T/NK-cell types) • Mixed phenotype acute leukemia, T-myeloid, NOS • lymphoma Classical Hodgkin lymphoma type PTLD • Provisional en+ty: natural killer (NK) cell lymphoblas+c • Anaplas+c large cell lymphoma, ALK- leukemia/lymphoma • Mycosis fungoides negave 5 6 1 3/21/16 Posi+on effect leads to new Pathophysiology Gene A promoter Gene B coding region expression rate • Chromosome Alteraons • Unbalanced: Change in total gene+c informaon Gene B coding region (addi+ons / dele+ons) Gene A coding region • Balanced: Gene+c informaon Del 17p (8%) rearranged Expression of trisomy 12 fusion genes with (12–16%) • DNA alteraons new funcon Del 11q • Single nucleode (18%) • Inser+ons Del 13q Breakpoint effect • Deleons (55%) CLL deletes coding • Duplicaons region and leads 7 to loss of func+on 8 Chromosomal Endogenous translocaons Growth Growth factor Exogenous factors receptors • Free radicals Gene • DNA replicaon errors • Ionizing radiaon amplificaon Signal Transcrip+on • V(D)J recombinaon • Chemicals transducers factors • Class switching Proto- • Drugs • Somac hypermutaon Retroviral oncogenes transduc+on Double Strand Breaks Inser+onal Gain of mutagenesis Funcon M G Point Oncogenes 1 mutaons Promote Cell Death G2 S DNA Repair proliferaon DNA Incorrect Neoplas+c cells in damaging peripheral blood event Genomic Instability Carcinogenesis Carcinogenesis Normal Cells in peripheral blood Bone marrow HSC pool 10 Adapted from: Clinical Laboratory Hematology, 3/E, McKenzie • Williams Growth Growth factor Oncogenes Promote factors receptors Lymphoproliferaon Signal Transcrip+on transducers factors Signal Transcrip+on Dimerizaon / Receptors Transducers Factors Transmembrane Ac+vaon Receptors (RTKs) BRAF NOTCH STAT3 BCL2 1. Receptor-growth factor BCR-ABL1 ETV6-RUNX1 BCR binding MYD88 BIRC3 TCF3-PBX1 2. Signal transduc+on JAK3 CD79A/B 3. Gene expression NOTCH MYC-IGH2 DNA RNA Protein 12 2 3/21/16 M G1 Growth Growth factor factors receptors Both alleles Cell Death G2 S DNA Repair must be affected Signal Transcrip+on Two-hit Incorrect transducers factors Inherited / hypothesis Sporadic Dimerizaon / Loss of Funcon Transmembrane Ac+vaon Tumor Receptors (RTKs) Suppressor Genes Inhibit proliferaon 1. Receptor-growth factor binding Carcinogenesis 2. Signal transduc+on 3. Gene expression DNA RNA Protein 13 Tumor Suppressors Inhibit Lymphoproliferaon Epigenecs • “On top of gene+cs” Signal Transcrip+on Epigenec • Informaon carried by the Transducers Factors Regulators genome that is not encoded Brooker by DNA ATM SF3B1 • DNA provides the gene+c plan P53 • Other factors affect how that plan will be expressed SF3B1 miR15/16 • DNA methylaon Brooker • Histone modificaons miRNA • Non-coding RNA (miRNA) 15 16 RNA interference Cancer Evolves by Natural Selec+on Depletes mRNA via (miRNA) sequence-specific pairing • Cancer is mul+ple different (sub-clonal) cancers occupying the same or dis+nct +ssue environments microRNA: Non-protein coding RNA Establishes and • Clonal expansion results due to compe++ve maintains ssue- advantage (proliferave or survival) over other (18 – 24 nucleo<des) binds mRNA specific expression sub-clones profile • Number of mutaons can vary from few (10–20) to hundreds of thousands • Most are ‘passengers’ (no effect on growth advantage) mRNA Translaonal • Others are func+onally relevant ‘drivers’ (confers degradaon inhibion selec+ve growth advantage) 18 17 3 3/21/16 Diffuse Walden- Chronic Hairy cell T-cell large Plasma cell T-cell acute Clonal Evolu+on is an Interplay of large B-cell ström lympho- leukemia granular myeloma lympho- lymphoma macro- cyc lympho- blasc Gene+c Changes: (DLBCL), globulin- leukemia cyc leukemia NOS emia leukemia CD79A 20% - - - - - - Selecvely • Survival/an+-apoptosis mutaon advantageous ‘driver’ CD79B 20% - - - - - - • Oncogenes mutaon lesions MYD88 30-75% 68-100% 2-10% - - - - mutaon Selec+vely neutral • Mono-alleleic loss NOTCH1 8% - 5-20% - - - >60% mutaon ‘passenger’ lesions • Tumor suppressors NOTCH2 8% - - - - - - mutaon BRAF 4% - - 99-100% - ??? - • Increase the rate of other gene+c changes mutaon ‘Mutator’ lesions - - - - • DNA repair genes SF3B1 - - 5-23% STAT3 3-5% - - - 28-40% 50% - P53 - - 10% - - 10% - Changes to the micro-environment miR 15a/ ??? - 55-66% - - Up to 50% - 19 16-1 20 MYD88 MYD88 • Toll-like receptors (TLR) are central to B-cell • MYD88 L265P missense clonal ‘driver’ receptor-independent an+gen response mutaon • Sense pathogen-associated molecular paerns • Iden+fied in 2011 in DLBCL (2012; WM) (bacteria, viruses, and fungi) • Gain of func+on to drive NF-kB • Ini+ates intracellular signaling (MYD88) Toll-interleukin-1 receptor domain DD TIR NF-kB IRAK interac+on L265P Promotes