Science Dialogue January 30 (Tues), 2007 at Niigata Prefectural Takada High School

Transcription

1 Science Dialogue January 30 (Tues), 2007 at Niigata Prefectural Takada High School Unique features of chronic myeloid leukemia in southern Vietnam (defoliants sprayed area) -- association with problem of dioxin pollution-- International Medical Center of Japan Research Institute, Division of Ultrafine Structure Phan Thi Xinh, Yuko Sato 1

2 Today s Talk 1. Dioxin and defoliant pollution caused by Vietnam War 2. What is chronic myeloid leukemia (CML)? 3. What prompted us to start this research? 4. Unique features of chronic myeloid leukemia in southern Vietnam 5. Epidemiologic study of dioxin pollution in Vietnam by Kanazawa University and our direction of further study 6. Dioxin pollution in Japan: in the past and the present 2

3 Today s Talk 1. Dioxin and defoliant pollution caused by Vietnam War 2. What is chronic myeloid leukemia (CML)? 3. What prompted us to start this research? 4. Unique features of chronic myeloid leukemia in southern Vietnam 5. Epidemiologic study of dioxin polution in Vietnam by Kanazawa University and our direction of further study 6. Dioxin pollution in Japan: in the past and the present 3

4 Dioxins--- 3 group (Toxicity is different) 1. polychlorinated dibenzo p-dioxins: PCDDs 7 kinds (75) 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD): Difficult to be decomposed in the environment, most toxic even a small amount 10 kinds (135) 2. polychlorinated dibenzofurans: PCDFs 3. coplanar polychlorinated biphenyls: Co-PCBs 12 kinds (29) 4

5 Physiologic effects of dioxins--1 Strong carcinogenic and teratogenic substance (most toxic among artificial substances) Generated in burning at around 300 (discomposed in burning at more than 800 ) Dioxins derived from agricultural chemicals: 4 times higher than that derived from waste incinerators Sensitivity of acute toxicity: exceedingly different among animals hamster (most sensitive) 1/800 of LD 50 guinea pig (most resistant) Human beings : resistant to dioxins however, to determine toxicity level (acute and chronic) is difficult Soluble in oil concentrated in human body through food chain a half life 7-12 years

6 Physiologic effects of dioxins--2 chronic toxicity High frequency of Reproduction abnormalities (infertility, abortion, stillbirth, congenital abnormalities, endometritis) Estrogen-like substance environmental hormone Most dangerous chemical in the environment 6

7 How to evaluate dioxins toxicity the toxicity of 2,3,7,8-TCDD = 1, and determine TEF toxic equivalency factor on each compound each compound x TEF TEQ (toxicity equivalency quantity We adapted WHO-TEF from 1997 in Japan. PCDDs TEF 2,3,7,8-TCDD 1 1,2,3,7,8-PeCDD 1 1,2,3,4,7,8-HxCDD 0.1 1,2,3,6,7,8-HxCDD 0.1 1,2,3,6,7,8-HxCDD 0.1 1,2,3,4,6,7,8-HpCDD 0.01 PCDFs TEF 2,3,7,8-TCDF 0.1 1,2,3,7,8-PeCDF ,3,4,7,8-PeCDF 0.5 1,2,3,4,7,8-HxCDF 0.1 1,2,3,6,7,8-HxCDF 0.1 1,2,3,7,8,9-HxCDF 0.1 7

8 Agent orange sprayed area in southern Vietnam Duration: Sprayed area >21,000 km 2 especially around US air-force base Sprayed volume 72x10 6 L (as dioxins:170 kg) Lat. 17 N 55% of mangrove area was destroyed (reported by VN 1080 division) 8

9 Today s Talk 1. Dioxin and defoliant pollution caused by Vietnam War 2. What is chronic myeloid leukemia (CML)? 3. What prompted us to start this research? 4. Unique features of chronic myeloid leukemia in southern Vietnam 5. Epidemiologic study of dioxin pollution in Vietnam by Kanazawa University and our direction of further study 6. Dioxin pollution in Japan: in the past and the present 9

10 Chronic Myeloid Leukemia: CML Increased WBC sometimes > 300,000/cmm 3 (WBC: 3,500-8,500/cmm 3 : normal value) WBC>50,000/cmm 3 splenomegaly WBC>70,000/cmm 3 general fatigue, weight loss, easy fatiguability, fever, abdominal fullness due to hepato-splenomegaly CML will be found at the health examination in Japan 10

11 Description of CML in old history In old Greek era: there is already a description of CML 1827 By Verbow the first accurate description of leukemic patients 63-year-old male: abdominal enlargement, fever, fatigue at autopsy, blood looked like wine-like pus 1835 by Donne 44-year-old male: splenomegaly, more than a half of blood was occupied with white blood cells Leukemia is a suppurated blood-disease associated with spleen disease rather than cancer Virchow R: an autopsy case showing giant splenomagaly and whitish blood Leuko (white) + Emia (blood) = Leukemia compound Latin 11

12 12

13 CML is a stem cell disorder Hematologic stem cells Ph Ph Ph chr. Gene rearrangements observed in CML Ph No.9 ABL gene der(9) tyrosine kinase activity(-) Ph chr. der(9) der(22) ABL/BCR gene ABL/BCR mrna (expressed in 70%) BCR/ABL gene BCR/ABL mrna tyrosine kinase activity(+++) 9q34: 22q11: No.22 BCR gene BCR/ABL protein (expressed in 100%) ABL gene BCR gene 13

14 Evidence of fusion of BCR gene and ABL gene No. 9 der( 9) der( 22) No.22 Cen ASS gene ABL gene 9q34 Intr.1 Int. 2 LSI ABL probe ( 650 kb) Tel LAMC3 gene nl(9) Cen GNAZgene BCR gene 22q11 M-BCR Tel LSI BCR probe ( 300 kb) nl(22) der(9) der(22) FISH study ABL gene BCR gene 14

15 2 types of Ph(-) CML No. 9 der( 9) No.22 der( 22) BCR/ABL fusion gene(+) The same as Ph(+)CML der( 22) No.22 der( 9) No. 9 BCR/ABL fusion gene(-) FISH, RT-PCR 15

16 der(9) deletion standard Ph trans. nl(22) nl(9) der(9) deletion It will occur when Ph trans. occurs nl(22) der(22) der(22) der(9) nl(9) der(9) the most powerful prognostic factor (tumor suppressor genes should be included in deleted region) 16

17 Typical clinical course of CML patients During CP, moderate clinical course W BC Only Ph chr. Chronic phase In 3-4 years, progress to more aggressive disease accelerated phase Acute phase Blasts +Ph +8 i(17q) Features of chronic phase Leukocytosis + basophilia hepatosplenomegaly Immature granulocytes < 5% Ph > 95%, additional chr. Abn.(-) Criteria of BC yrs myeloblasts >= 5%, promyeloblasts >= 20% additional chr. Abnormalities (+Ph,+8,iso(17q),+19,+21) lymph node enlargement, blastoma, treatment-resistant splenomegaly 17

18 Typical clinical course of CML patients During CP, moderate clinical course W BC Only Ph chr. Chronic phase +Ph In 3-4 years, progress to more aggressive disease +8 i(17q) accelerated phase Acute phase Blasts yrs +Ph +8 i(17q) Typical additional chromosome abnormalities 18

19 Today s Talk 1. Dioxin and defoliant pollution caused by Vietnam War 2. What is chronic myeloid leukemia (CML)? 3. What prompted us to start this research? 4. Unique features of chronic myeloid leukemia in southern Vietnam 5. Epidemiologic study of dioxin pollution in Vietnam by Kanazawa University and our direction of further study 6. Dioxin pollution in Japan: in the past and the present 19

20 Prognosis comparison of CML patients with hydroxyurea treatment Southern VN (Tuan TQ. J : Vietnam Med 2002; 5: 9-12) Northern VN (Thanh NH: in personal communication) (Hehlmann H et al., Blood 1993; 82: ) France (The Benelux CML study group. Blood 1998; 91: ) No. of patients analyzed Median survival (mo) month survival (%) Prognosis of CML patients in southern Vietnam is extremely bad. Compared with that in northern Vietnam 20

21 1. Why is the prognosis of CML patients in southern Vietnam so bad? 2. Are there specific molecular and cytogenetic features in southern Vietnamese CML patients? 3. If any, are they associated with damage of dioxin pollution during Vietnam War? Molecular and cytogenetic study of southern Vietnam CML ----Collaborated study with Blood transfusion and Hematology Hospital at Ho Chi Minh City--- 21

22 Today s Talk 1. Dioxin and defoliant pollution caused by Vietnam War 2. What is chronic myeloid leukemia (CML)? 3. What prompted us to start this research? 4. Unique features of chronic myeloid leukemia in southern Vietnam 5. Epidemiologic study of dioxin pollution e in Vietnam by Kanazawa University and our direction of further study 6. Dioxin pollution in Japan: in the past and the present 22

23 Materials and Methods 47 patients BM or PB 1) at diagnosis: 32 2) in chronic phase: 10 3) in accelerated phase : 5 47 FISH study Chromosome study M Patient 45 Patient 46 Patient 47 Patient 42 HT93A N K562 RT-PCR study b3a2 (287 bp) b2a2 (212 bp) ABL inner control (160 bp) 23

24 Chromosome study 1. Frequency of Ph(+) and Ph(-) CML 2. Karyotype analysis 24

25 Results of chromosome study--1 Ph(+) CML: 44/47 (93.6%) Ph (-) CML: 3/47 (6.4%) 2 types of Ph(-) CML Frequencies of Ph(-) CML China: 5.0% Canada: 5.0% India: 5.0% Japan: 0.5% Gene rearrangements in Ph(-) BCR/ABL(+) CML 50%--BCR/ABL(+) the same disease as Ph(+)CML 50%--BCR/ABL(-) a different disease from Ph(+)CML, similar to MDS/MPD? Detection by FISH, RT-PCR 25

26 Clinical and cytogenetic findings of 3 Ph(-) CML patients Pt. No Age/ Sex Phase obtained sample (mo. after diagnosis) Source WBC counts at diag. (x10 9 /L) WBC counts at sampling (x10 9 /L) Platelet counts at diag. (x10 9 /L) Monocytosis at diag. Cytogenetic finding FISH finding of BCR/ABL Major BCR/ABL mrna ABL/BCR mrna 45 40F CP (11) BM ,XX (+) Type 2 b2a2 & b3a2 (-) 46 75F CP (23) BM ,XX (-) b2a2 1b-b M Dx (0) BM ,XY (-) b3a2 1b-b4 Ph(-) no BCR/ABL FISH signals 15 metaphase 200 interphase cells BCR/ABL transcripts (+) 2 clones Major clone BCR/ABL(-) Minor clone < 1/200 (0.5%) BCR/ABL(+) 26

27 Result of chromosome study-2 a) at Dx: 4/10 (40.0%) b) in CP: 11/32 (34.4%) c) in AP: 2/ 5 (40.0%) 17/47 (36.2%) } among CP-patients 15/42 (35.7%) Completely different abnormalities from additional abnormalities such as +Ph, +8, +i(17q), +19, +21 trisomy 13 add(1)(p36) +add(1)(p31) del(3)(p21p23) add(6)(p22) del(7)(p14p15) add(10)(p13) add(11)(p15) Xinh PT, et al. Cancer Genet Cytogenet 168:59-68,

28 Southern Vietnam Other countries In chronic phase: 15/42 (35.7%) In chronic phase: % (Anastasi et al., Leukemia, 1995;9: ) In blastic phase: about 80% double Ph, +8, iso(17q), +19 and +21 (Kantarjian et al., Blood, 1993;82: ) Among CML patients in southern Vietnam, previously unreported chromosome abnormalities were found in chronic phase with high frequency 28

29 FISH study 1. Analysis of BCR/ABL breakpoints 2. The frequency of der(9) deletion 29

30 In the same patient, cells with standard Ph translocation and cells with der(9) deletion were found. der(9) deletion A B nl(9) der(9) C der(9) standard Ph transl. nl(9) der(22) der(22) nl(22) nl(22) Xinh PT, et al. Cancer TGenet Cytogenet 164: , T

31 There are 2 clones Pt. 3 Pt. 5 1) Cells with standard Ph translocation ) Cells with der(9) deletion 3 4 der(9) deletion Ph translocation clones der(9) deletion The first report which showed that der(9) deletion occurs in the progression of disease. (Xinh PT, et al. Cancer Genet Cytogenet 164: ,

32 Summary 1. Unique chromosomal abnormalities were found with high frequency at diagnosis: 4/10 (40.0%) in chronic phase: 11/32 (34.4%) 2. der(9) deletion was found with high frequency: 11/47(23.4%) 3. 2 kinds of clones in the same patient: 1) a clone with der(9) deletion and one without it 2) a clone with BCR/ABL gene and one without it Is there the existence of genomic instability? 32

33 What is the cause of inducing genomic instability? Influences of dioxins pollution 33

34 Today s Talk 1. Dioxin and defoliant pollution caused by Vietnam War 2. What is chronic myeloid leukemia (CML)? 3. What prompted us to start this research? 4. Unique features of chronic myeloid leukemia in southern Vietnam 5. Epidemiologic study of dioxins pollution in Vietnam by Kanazawa University and our direction of further study 6. Dioxins pollution in Japan: in the past and the present 34

35 Long term effects of war agent orange/dioxins on human health of people who have lived in a sprayed area Selected about 4100 individuals from 2 areas with similar life style on both sides of Lat. 17 N non-sprayed area (Cam Phuc commune northern VN) heavily sprayed area (Cam Chinh commune southern VN Lat. 17 N A large scaled endemic study (stillbirth, congenital abnormalities, birth rate of boy and girl, nutrition condition) Measurement of dioxins in the environment soil, water, domestic fowl and pig) Measurement of dioxins in human tissues (serum, adipose tissue, and breast milk) from individuals Even now (30 years after Vietnam War), individuals in a heavily sprayed area showed 3-4 times higher level of serum dioxin compared with individuals in a non-sprayed area. However, some individuals showed low dioxins level. 35

36 Measurement of dioxins by Prof. Teruhiko Kido (Kanazawa University and Kanazawa Medical University) Using 5 ml serum or 10 ml breast milk, 6 kinds of dioxins are measurable Highest detection level in the world 36

37 Next Plan High dioxin group Low dioxin group Sprayed area 3 Control Non-sprayed area Sister chromatid exchange 1. Chromosome analysis of T cells 2. Sister chromatid exchange (SCE) analysis 3. P450(CYP2D6) analysis 37

38 Next Plan-2 1. Among people with high dioxins value: Do they show chromosomal abnormalities? Do they show high SCE index? 2. Among people with low dioxins value: Do they show specific P450 subtype? 38

39 Today s Talk 1. Dioxin and defoliant pollution caused by Vietnam War 2. What is chronic myeloid leukemia (CML)? 3. What prompted us to start this research? 4. Unique features of chronic myeloid leukemia in southern Vietnam 5. Epidemiologic study of dioxin pollution in Vietnam by Kanazawa University and our direction of further study 6. Dioxins pollution in Japan: in the past and the present 39

40 Dioxin pollution in Japan Damages by defoliants used during the Vietnam War make an issue Dioxins were detected in ash of incinerators 74% of garbage were burned in incinerators 2000 incinerators in the whole country produce 17kg dioxin a year, which is 1/10 of a total amount of dioxins used in Vietnam War. A total amount of agricultural chemicals used for 30 years since 1960s is equal to a total amount of dioxins used in the Vietnam War Guideline for prevention of dioxin generation Designate dioxin as a substance for Law of Prevention for Air Pollution Principle guideline for measures against dioxin Goal: within 4 years, 90% reduction compared with the value of

41 Dioxin pollution in Japan--2 Change of dioxins discharge from incinerators - 98% : special law of measurement against dioxins : come into force of the law : complete enforcement 2003 year discharge decreased to 145 g industrial about 74 g, domestic about 71 g) Problems to be solved in the future 1. Measure dioxin discharge from small incinerators 2. Close and improve inappropriate final treatment of industrial waste 3. Scrap incinerators smoothly 41

42 Changes of dixions discharge and dixions level in air and water in Japan borrowed from Prof. Kido, Kanazawa Univ

43 Change of daily intake of dioxins borrowed from Prof. Kido, Kanazawa Univ. 43

44 Change of dioxins level in human breast milk borrowed from Prof. Kido, Kanazawa Univ. Still higher than that of control group in southern Vietnam borrowed from Prof. Kido, Kanazawa Univ. 44

45 Daily intake of dioxins/person in Japan WHO TDI borrowed from Prof. Kido, Kanazawa Univ. TEQ: toxicity equivalecy quantity TEF: toxic equivalecy factor 45

46 Thank you for your attention 46