Transcript 혈액종양의 분자세포유전학 Back to Basic Molecular Cytogenetics in hematologic Malignancy

혈액종양의 분자세포유전학Back to Basic
Molecular Cytogenetics
in hematologic Malignancy
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Historical Backgroud
1960 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)
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Cytogenetic & Gene Rearrangement
1980
1983
1984
1990
1991
1993
1997
MYC-IGH t(8;14)
BCR-ABL1 t(9;22)
IGH-BCL2 t(14;18)
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)
CBFB-MYH11
inv(16)
MLL-MLLT3
t(9;11)
TEL-AML1
t(12;22)
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Cytogenetic Studies
for Hematologic Malignancies in Korea
Table 4. Karyotypic pattern
in cases with CML
Karyotype
t(22q;9q)
22q22q-. -9
49,XXX,+9,+21
Normal
KimSW, et al.: Banding patterns
of normal human chromosomes.
The Seoul Journal of Medicine
21(2):133-137(1980).
No.
7
6
1
1
6
21
조혈기질환에 있어서 골수세포
염색체 분석에 관한 연구.
대한혈액학회지 23(2),1988
비밀
가을동화
안녕 내사랑
태양은 가득히
Last Concert
Love Story
Dying Young
Marvin’s Room
Roman Holiday
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)
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Philadelphia Chromosome (=Ph)
der(22)t(9;22)(q34.1;q11.21)
q11.2
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.
CML Blast Crisis - Karyotype
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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
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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 curableBack to Basic
PML
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)
RARA
PML/RARA
AML-M4Eo with inv(16)
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 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
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 노동자계층
의 여자가 백혈병으로 시한부 인생
을 사는 상류계층의 청년을 간호하
기 위해 고용된다. 그리고 둘은 사랑
에 빠진다.
KBS 미니시리즈
가을동화
극중의 의사는 은서가 백혈병에 걸린 사실을 확인
하는 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
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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
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




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
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MDS
Normal Chromosome
del 5q
-7, del 7q
del 9q
del 20q
+8
complex defects
t(1;3), t(2;11)
AML
t(3;3), inv(3)
t(6;9), inv(16)
t(8;21)
t(9;22)
t(v;11)
t(15;17)
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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 아
버지의 사랑을 그린 작품
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Cytogenetics in
Acute Lymphoblastic Leukemia
Poorly spread, short
Fuzzy chromosomes
Indistinct bands
Low success rate
50% in conventional culture
76% Clarkson (1985)
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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 Back to Basic
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
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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.
Clinical Application of FISH
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 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.
The first is to help yourself, the second is to help others."
Audrey Hepburn
1929-1993
Solid Tumor Cytogenetics
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1. Preparation of tumor sample
2. Tumor cell culture
Culture Media
FBS (11-20%)
Insulin
Antibiotics
Transferrin
Additives
Hydrocortisone
Tumor cells
EDGF
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
Solid Tumor Abnormalities
der(1)
del(1p)
del(3)(p12p14)
del(3)(p14p23)
del(3p)
del(7)(q21q31)
del(9)(p13)
del (11)(p13)
del(22q) or -22
i(6)(p10)
i(8)(q10)
i(12)(p10)
i(17)(q10)
inv(10)(q11q21)
t(1;17)(p36;q12)
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breast carcinomas
colorectal adenoma
breast carcinomas
small cell carcinoma of lungs
renal cell carcinoma,
non-small cell lung cancer
uterine leiomyomas
malignant melanoma
Wilm's tumor
meningiomas
retinoblastoma
uveal melanoma
testicular germ cell tumors
primitive neuroectodermal tumor
papillary carcimoma of the thyroid
neuroblastoma
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inv(10)(q11q21)
t(1;17)(p36;q12)
t(2;13)(q35;q14)
t(3;8)(p21;q12)
t(3;12)(q27-28;q13-15)
t(9;22)(q22;q11-12)
papillary carcimoma of the thyroid
neuroblastoma
alveolar rhabdomyosarcoma
pleomorphic ademoma of salivary glands
sporadic typical lipomas
extraskeletal myxoidchondrosarcoma
mucoepidermoid carcinoma,
t(11;19)(q21;p11)
adenolymphoma of salivary gland
t(11;22)(p13;q12)
intraabdominal desmoplastic small round cell tumor
Askins tumor,
t(11;22)(q24;q12)
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
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25000
20000
Solid Tumors
Malig Lymphoma
Hemat Malignancy
15000
10000
5000
0
1983 1985 1988 1991 1994
“Catalog of Chromosome Aberrations in Cancer ”