Donor single nucleotide polymorphism in ACAT1 affects the incidence of graft-versus-host disease after bone marrow transplantation

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Figure legends

Figure 1

Comparison of the expression level of ACAT1 mRNA in T cells among rs11545566

genotypes. The normalized expression level of ACAT1 of the AA genotype was used as

a reference. The relative expression levels of ACAT1 in (a) CD8-positive T cells and (b)

CD4-positive T cells were compared among three rs11545566 genotypes (AA, n = 3;

AG, n = 4; GG, n = 3) using one-way ANOVA (left) and between the AA (n = 3) and

non-AA (n = 7) genotypes using a t-test (right). Data of two independent experiments

(mean ± S.E.M.) each performed in triplicate are shown.

Figure 2

Impact of donor ACAT1 rs11545566 genotype on the incidence of acute GVHD. (a)

Cumulative incidences of grade II-IV acute GVHD in patients transplanted from donors

with ACAT1 rs11545566 AA (solid line, n = 36), AG (dashed line, n = 52), and GG

(dotted line, n = 28) genotypes are shown. (b) Cumulative incidences of grade II-IV

acute GVHD in patients transplanted from donors with ACAT1 rs11545566 AA (solid

line, n = 36) and non-AA (dashed line, n = 80) genotypes are shown.

Figure 3

Impact of donor ACAT1 rs11545566 genotype on the incidence of chronic GVHD. (a)

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Cumulative incidences of extensive chronic GVHD in patients transplanted from donors

with ACAT1 rs11545566 AA (solid line, n = 36), AG (dashed line, n = 49), and GG

(dotted line, n = 25) genotypes are shown. (b) Cumulative incidences of extensive

chronic GVHD transplanted from donors with ACAT1 rs11545566 AA (solid line, n =

36) and non-AA (dashed line, n = 74) genotypes are shown.

27

Table 1 Patient characteristics categorized by ACAT1 rs11545566 genotype of the donors

Donor genotype

Number of patients

Median age (range), y

P-value

AA

AG

GG

36

52

28

35 (16 − 58) 36 (15 − 51) 35 (16 − 56)

Age at transplantation, n (%)

15 − 20 y

5 (14)

8 (15)

4 (14)

21 − 30 y

5 (14)

11 (21)

7 (25)

31 − 40 y

12 (33)

16 (31)

8 (29)

41 − 50 y

9 (25)

15 (29)

6 (21)

51 − 58 y

5 (14)

2 (4)

3 (11)

Male

23 (64)

32 (62)

12 (43)

Female

13 (36)

20 (38)

16 (57)

Male patient from female donor

12 (33)

17 (33)

5 (18)

Other combinations

24 (67)

35 (67)

23 (82)

Acute myeloid leukemia

12 (33)

16 (31)

7 (25)

Acute lymphoblastic leukemia

5 (14)

8 (15)

6 (21)

Chronic myeloid luekemia

9 (25)

18 (34)

7 (25)

Myelodysplastic syndrome

6 (17)

2 (4)

2 (7)

0 (0)

1 (2)

1 (4)

Non-malignancies

Disease risk, n (%)

4 (11)

7 (14)

5 (18)

Standard-risk

28 (78)

41 (79)

20 (71)

High-risk

8 (22)

11 (21)

8 (29)

Positive

26 (72)

45 (87)

22 (79)

Negative

6 (17)

2 (4)

4 (14)

Unknown

4 (11)

5 (10)

2 (7)

MAC

35 (97)

50 (96)

25 (89)

RIC

1 (3)

2 (4)

3 (11)

0.83

Sex, n (%)

0.18

Sex mismatch between patient and donor, n (%)

0.31

Diagnosis, n (%)

#1

Other malignancies

#2

0.65

0.74

Cytomegalovirus serostatus, n (%)

0.31

Conditioning regimen, n (%)

0.31

#1 Other malignancies include multiple myeloma and mantle cell lymphoma.

#2 Non-malignancies include aplastic anemia, pure red cell aplasia, and paroxysmal nocturnal hemoglobinuria.

Table 2 Multivariate analysis of risk factors for transplant outcome

Outcome and significant factor

Hazard ratio (95% confidence interval)

P-value

Grade II-IV acute GVHD

Age

Donor rs11545566 genotype

<35 y

≥35 y

7.14 (1.58 − 32.2)

AA

Non-AA

3.70 (0.88 − 15.6)

Standard-risk

High-risk

3.12 (1.29 − 7.74)

AA

Non-AA

5.62 (1.29 − 24.5)

Standard-risk

High-risk

2.67 (1.22 − 5.85)

<35 y

≥35 y

3.45 (1.18 − 10.0)

<35 y

≥35 y

2.25 (1.17 − 4.33)

Standard-risk

High-risk

2.51 (1.33 − 4.19)

0 or I

II − IV

2.08 (1.03 − 4.19)

0.011

0.079

Extensive chronic GVHD

Disease risk

Donor rs11545566 genotype

0.014

0.021

Relapse rate

Disease risk

0.014

NRM

Age

0.023

OS rate

Age

Disease risk

Acute GVHD

0.015

0.004

0.038

Figure 1 (a) P = 0.098 1.0 0.5 1.5 Relative expression level Relative expression level 1.5 P = 0.027 1.0 0.5 0.0 0.0 AA AG GG AA non-AA (b) P = 0.025 1.0 0.5 1.5 Relative expression level Relative expression level 1.5 P = 0.098 1.0 0.5 0.0 0.0 AA AG GG AA non-AA Figure 2 (a) Cumulative incidence of

grade II-IV acute GVHD

100 P = 0.169 80 60 40 20 0 0 20 60 80 40 Days after transplantation 100 (b) 100 Cumulative incidence of

grade II-IV acute GVHD

P = 0.06 80 60 40 20 0 0 20 60 80 40 Days after transplantation 100 Figure 3 (a) Cumulative incidence of

extensive chronic GVHD

100 P = 0.022 80 60 40 20 0 0 0.5 1.5 2.0 1.0 Years after transplantation (b) Cumulative incidence of

extensive chronic GVHD

100 P = 0.008 80 60 40 20 0 0 0.5 1.5 2.0 1.0 Years after transplantation ...