혈액종양의 분자세포유전학 Molecular Cytogenetics in hematologic Malignancy

Historical Backgroud Philadelphia chromosome (Ph) 1970 Banding technique 1970 t(9;22) in CML 1973 t(8;21) in AML-M2 t(8;14) in Burkitt lymphoma/Leukemia 1977 t(15;17) in AML-M3, t(4;11) 1979 High-resolution banding technique 1980 1982 t(9;11) in AML-M5a, inv(3) in AML 1983 inv(16) in AML-M4E 1984 t(1;19) in ALL t(1;3) in AML with dysmegakaryopoiesis 1990 Molecular cytogenetics (FISH, CGH)

Cytogenetic & Gene Rearrangement 1980 1983 MYC-IGH t(8;14) 1984 BCR-ABL1 t(9;22) IGH-BCL2 t(14;18) 1990 1991 MLL-AFF1 t(4;11) E2A-PBX1 t(1;19) PML-RARA t(15;17) RUNX1-RUNX1T1 t(8;21) DEK-NUP214 t(6;9) 1993 CBFB-MYH11 inv(16) MLL-MLLT3 t(9;11) 1997 TEL-AML1 t(12;22)

Cytogenetic Studies for Hematologic Malignancies in Korea Table 4. Karyotypic pattern in cases with CML Karyotype No. t(22q;9q) 7 22q- 6 22q-. -9 1 49,XXX,+9,+21 Normal 21 조혈기질환에 있어서 골수세포 염색체 분석에 관한 연구. 대한혈액학회지 23(2),1988 KimSW, et al.: Banding patterns of normal human chromosomes. The Seoul Journal of Medicine 21(2):133-137(1980).

Leukemia in Cinema 비밀 가을동화 안녕 내사랑 태양은 가득히 Last Concert Love Story Dying Young Marvin’s Room Roman Holiday

having to say you're sorry Love means never having to say you're sorry Starring Ali MacGraw (Jeniffer) Ryan O'Neal (Oliver) Music (Snow Frolic) Francis Lai Written by Erich Segal (1970)

Philadelphia Chromosome (=Ph) der(22)t(9;22)(q34.1;q11.21) BCR q34 ABL Ph (formerly Ph1) may be used in text, but not in the description of the karyotype, where der(22)t(9;22)(q34;q11.2) is recommended.

50,XX,+8,t(9;22)(q34;q11.2),+10,+19, +der(22)t(9;22)(q34.1;q11.21) CML Blast Crisis - Karyotype 50,XX,+8,t(9;22)(q34;q11.2),+10,+19, +der(22)t(9;22)(q34.1;q11.21)

Practical Use of Cytogenetics in CML Advantages of Cytogentics, compared to molecular DNA studies for BCR gene rearrangement Distinguish betwn variant Ph and standard t(9;22) Dectect other abnormalities : +8, i(17q),+Ph,+19 Predict or confirm blast crisis Give information regarding percentage of normal vs abnormal cells Valuable after BMT to follow engraftment of the donor cells and identify possible relapse

Dedicato A Una Stella 1976, Italy Richard Johnson Pamela Villoresi Stelvio Cipriani 불치 병(백혈병)에 걸린 아가씨와 오랜 슬럼프에 빠진 피아니스트의 슬픈 사랑을 엮은 러브스토리

APL with t(15;17)(q22;q21), PML/RARA From highly fatal to highly curable RARA PML PML/RARA G-banding : t(15;17)(q22;q21) & Variants of t(15;Var;17), t(11;17), t(5;17) RT-PCR, FISH : PML/RARA fusion transcript ATRA (all-trans retinoic acid) & ATO (arsenic trioxide) 11

AML-M4Eo with inv(16) Young patients, organomegaly, abnormal eosinophils Specifically associated with M4Eo in over 50% of cases Favorable prognosis, High incidence of CNS relapse MYH11 CBFB CBFB-MYH11

It’s not just a love story. 사랑을 위하여 (1991) It’s not just a love story. It’s a life story Starring Julia Roberts Campbell Scott Music by Kenneth Gorelick 자유분방한 사고를 가진 노동자계층의 여자가 백혈병으로 시한부 인생을 사는 상류계층의 청년을 간호하기 위해 고용된다. 그리고 둘은 사랑에 빠진다.

KBS 미니시리즈 가을동화 극중의 의사는 은서가 백혈병에 걸린 사실을 확인하는 과정에서 ‘유전이 될 확률이 높은 병’이라고 말하는데…… 하지만 백혈병이 유전되는 병은 아니다. 윤준서 (송승헌) 윤은서 (송혜교) 한태석 (원 빈)

Acute Myeloid Leukemia and Related Neoplasms Proposed WHO Classification AML with recurrent genetic abnormalities t(8;21), RUNX1-RUNX1T1 t(15;17), PML/RAR inv(16), CBF/MYH11 t(9;11), MLLT3/MLL t(6;9), DEK/NUP214  inv(3) or t(3;3), RPN1-EVI1  t(1;22), RBM15-MKL1  Acute myeloid leukemia with myelodysplasia-related changes Therapy-related myeloid neoplasms Acute myeloid leukemia, not otherwise specified Myeloid sarcoma Myeloid proliferations related to Down syndrome Blastic plasmacytoid dendritic cell neoplasm

MDS AML Normal Chromosome del 5q -7, del 7q del 9q del 20q +8 complex defects t(1;3), t(2;11) MDS t(3;3), inv(3) t(6;9), inv(16) t(8;21) t(9;22) t(v;11) t(15;17) AML

Myeloid Malignancy Secondary to Radiotherapy or Chemotherapy Clinical features AML-M1,M2,M6 Lower remission rate and long-term reponse Cytogenetics Usually absence of t(8;21), t(15;17) & inv(16) Usually complex karyotype Common abnormalities: -5, del(5q) in chemotherapy, -7, del(7q) in radiation therapy, Abnormalities of 3p, 11q23, 12p, and 17

José Carreras

José Carreras International Leukaemia Foundation José Carreras E. Donald Thomas Seattle's Hutchinson Clinic following a benefit concert in San Diego. Dinner party after E. Donnall Thomas’ lecture in Kyoto (1991)

MBC 드라마 2000.12.8-9 원작 : 조창인 출연 : 정보석, 박지영 급성림프아구성백혈병 환자인 아들에게 자신의 모든 것을 희생하며 살려내는 아버지의 사랑을 그린 작품

Cytogenetics in Acute Lymphoblastic Leukemia Poorly spread, short Fuzzy chromosomes Indistinct bands Low success rate 50% in conventional culture 76% Clarkson (1985)

Ploidy Groups in Childhood ALL Hyperdiploid >50 28% Early pre-B immunophenotype Found at the age of 2-10 years Lower WBC count, Favorable prognosis Hyperdiploid 47-49 13% Diploid 46 9% Pseudodiploid 46 38% Hypodiploid <46 6%

Precursor Lymphoid Neoplasm Proposed WHO Classification B lymphoblastic leukemia/lymphoma, NOS B lymphoblastic leukemia/lymphoma with recurrent genetic abnormalities t(9;22)(q34;q11.2); BCR-ABL1 t(v;11q23); MLL rearranged t(12;21)(p13;q22) TEL-AML1 (ETV6-RUNX1) hyperdiploidy hypodiploidy t(5;14)(q31;q32) IL3-IGH t(1;19)(q23;p13.3);TCF3-PBX1 T lymphoblastic leukemia/lymphoma

Jacqueline Kennedy Onassis 1929-1994

Cytogenetics Technologies Karyotyping Provides a visual examination of the entire genome the best coverage but not the best resolution (≥ 5 Mb) Banding resolution differs from preparation to preparation FISH Sensitive (high resolution) Only provides information on tested regions, other aberrations will NOT be tested, i.e. not genome view M-FISH(Multi-FISH, Spectral karyotyping) Array-based Copy Number / CGH Analysis Single step global genome scan prevents FISHing expedition BAC (Bacterial Artificial Chromosome) Array Requires well characterized and high resolution clones High Resolution Oligonucleotide Microarray Highest resolution. Precise identification of gains and losses of genetic material. 25

Clinical Application of FISH Microdeletion syndromes Prader-Willi/Angelman syndrome DiGeorge/CATCH 22 syndrome Williams syndrome etc. Marker chromosome Hematologic malignancies t(9;22): abl/bcr t(15;17): PML/RARa 11q23: MLL t(12;21): TEL/AML1 BMT follow-up using X/Y

46,XY,t(9;22)(q34;q11.2)

"Remember you another hand "Remember you another hand. The first is to help yourself, the second is to help others." Audrey Hepburn 1929-1993

Solid Tumor Cytogenetics 1. Preparation of tumor sample 2. Tumor cell culture Culture Media FBS (11-20%) Antibiotics Additives Tumor cells 2. Harvest 1) Colcemid (0.01-0.1mcg/mL) treatment 2) Hypotonic (0.050-0.075M KCl) treatment 3) Fixation 4) Slide preparation 3. Staining and analysis Insulin Transferrin Hydrocortisone EDGF

Solid Tumor Abnormalities der(1) breast carcinomas del(1p) colorectal adenoma del(3)(p12p14) del(3)(p14p23) small cell carcinoma of lungs del(3p) renal cell carcinoma, non-small cell lung cancer del(7)(q21q31) uterine leiomyomas del(9)(p13) malignant melanoma del (11)(p13) Wilm's tumor del(22q) or -22 meningiomas i(6)(p10) retinoblastoma i(8)(q10) uveal melanoma i(12)(p10) testicular germ cell tumors i(17)(q10) primitive neuroectodermal tumor inv(10)(q11q21) papillary carcimoma of the thyroid t(1;17)(p36;q12) neuroblastoma

inv(10)(q11q21) papillary carcimoma of the thyroid t(1;17)(p36;q12) neuroblastoma t(2;13)(q35;q14) alveolar rhabdomyosarcoma t(3;8)(p21;q12) pleomorphic ademoma of salivary glands t(3;12)(q27-28;q13-15) sporadic typical lipomas t(9;22)(q22;q11-12) extraskeletal myxoidchondrosarcoma t(11;19)(q21;p11) mucoepidermoid carcinoma, adenolymphoma of salivary gland t(11;22)(p13;q12) intraabdominal desmoplastic small round cell tumor t(11;22)(q24;q12) Askins tumor, Ewings sarcoma, esthesioneuroblastoma t(12;14)(q14-15;q23-24) uterine leiomyomas t(12;16)(q13;p11.2) myxoid liposarcoma t(X;1)(p11;q21) malignant papillary renal cell carcinoma t(X;18)(p11.2;q11.2) synovial sarcoma

“Catalog of Chromosome Aberrations in Cancer ”