Banini et al Haptoglobin and Vitamin E 1
Supplementary Table 1. Baseline characteristics at enrollment of adult patients by haptoglobin genotype
Haptoglobin genotype
1Hp 2-2 Hp 2-1 Hp 1-1 P
2Characteristics N=51 N=68 N=16
Treatment 0·26
Vitamin E 29 (57%) 30 (44%) 10 (63%)
Placebo 22 (43%) 38 (56%) 6 (38%)
Demographic:
Age 47·1 (11·4) 47·7 (11·1) 49·4 (10·4) 0·77
Sex: male 20 (39%) 21 (31%) 7 (44%) 0·48
Ethnicity: Hispanic 9 (18%) 3 (4%) 2 (13%) 0·06
Race: 0·74
White 40 (80%) 58 (87%) 14 (93%)
Black 1 (2%) 2 (3%) 0 (0%)
Other
39 (18%) 7 (10%) 1 (7%)
Anthropometric:
BMI (kg/m
2) 34·7 (7·4) 33·9 (6·8) 35·8 (5·7) 0·56
Laboratory measures:
Glucose (mmol/L) 5·30 (0·79) 5·27 (0·81) 5·25 (0·74) 0·96
HOMA-IR 28·7 (19·6) 32·1 (25·8) 32·5 (14·5) 0·69
Triglycerides (mmol/L) 1·90 (0·93) 1·87 (1·16) 1·67 (0·48) 0·73
Cholesterol, total (mmol/L) 5·34 (0·94) 4·87 (0·99) 5·17 (1·00) 0·03
Cholesterol, HDL (mmol/L) 1·15 (0·26) 1·12 (0·28) 1·13 (0·34) 0·81
Alanine aminotransferase (U/L) 82·4 (52·1) 75·1 (47·7) 90·1 (35·4) 0·47 Asparate aminotransferase (U/L) 56·5 (33·3) 56·9 (34·1) 58·1 (21·6) 0·98
Alkaline phosphatase (U/L) 78·0 (25·1) 80·6 (26·8) 88·6 (32·3) 0·40
γ-glutamyl transpeptidase (U/L) 61·3 (49·6) 49·9 (33·6) 48·9 (24·3) 0·26 Histological
Definite steatohepatitis 37 (73%) 53 (78%) 14 (88%) 0·48
Total NAFLD activity score (0-8) 5·1 (1·4) 4·8 (1·5) 4·8 (1·3) 0·45
Steatosis grade (0-3) 1·9 (0·7) 1·9 (0·8) 1·7 (0·9) 0·55
Lobular inflammation score (0-3) 1·8 (0·7) 1·6 (0·7) 1·7 (0·7) 0·24
Hepatocellular ballooning score (0-2) 1·4 (0·7) 1·3 (0·7) 1·4 (0·7) 0·98
Fibrosis stage (0-4) 1·4 (1·2) 1·6 (1·1) 1·7 (0·8) 0·54
1 Adult patients are from the PIVENS trial with paired baseline and end of treatment biopsies and DNA. Total number of patients is 135.
Data are No. (%) or mean (SD).
2 P-value calculated using Fisher’s exact test for categorical characteristics and ANOVA for continuous characteristics.
3 3 patients refused to report a race; however, 2 of the 3 identified as Hispanic.
Abbreviations: BMI, body mass index; HDL, high density lipoprotein; HOMA-IR, homeostasis model assessment-estimated insulin resistance; Hp, haptoglobin; NAFLD, nonalcoholic fatty liver disease
Banini et al Haptoglobin and Vitamin E 2
Supplementary Table 2. Baseline characteristics at enrollment of pediatric patients by haptoglobin genotype
Haptoglobin genotype
1Hp 2-2 Hp 2-1 Hp 1-1 P
2Characteristics N=27 N=45 N=21
Treatment 0·96
Vitamin E 14 (52%) 24 (53%) 10 (48%)
Placebo 13 (48%) 21 (47%) 11 (52%)
Demographic:
Age 12·6 (2·8) 12·8 (2·3) 12·0 (2·4) 0·48
Sex: male 22 (81%) 35 (78%) 18 (86%) 0·79
Ethnicity: Hispanic 15 (56%) 33 (73%) 19 (90%) 0·03
Race: 0·06
White 24 (92%) 31 (76%) 15 (75%)
Black 1 (4%) 0 (0%) 0 (0%)
Other
31 (4%) 10 (24%) 5 (25%)
Anthropometric:
BMI (kg/m
2) 32·6 (5·5) 34·2 (6·0) 32·9 (5·6) 0·48
Laboratory measures:
Glucose (mmol/L) 4·91 (0·50) 4·83 (·53) 5·04 (0·36) 0·25
HOMA-IR 47·3 (31·3) 69·5 (113·9) 57·6 (68·1) 0·58
Triglycerides (mmol/L) 1·63 (0·93) 1·75 (1·17) 1·32 (0·72) 0·27
Cholesterol, total (mmol/L) 4·52 (1·13) 4·61 (0·78) 4·44 (0·70) 0·76
Cholesterol, HDL (mmol/L) 0·99 (0·34) 0·98 (0·23) 0·96 (0·17) 0·92
Alanine aminotransferase (U/L) 126·0 (70·3) 126·1 (64·7) 115·8 (68·4) 0·84 Asparate aminotransferase (U/L) 74·2 (45·7) 73·0 (30·8) 72·2 (51·6) 0·99 Alkaline phosphatase (U/L) 248·4 (103·0) 218·4 (95·4) 205·0 (70·3) 0·24 γ-glutamyl transpeptidase (U/L) 40·1 (16·9) 53·9 (34·2) 55·5 (30·2) 0·11 Histological
Definite steatohepatitis 12 (44%) 15 (33%) 12 (57%) 0·19
Total NAFLD activity score (0-8) 4·8 (1·4) 4·6 (1·4) 5·1 (1·8) 0·42
Steatosis grade (0-3) 2·3 (0·8) 2·2 (0·8) 2·3 (0·9) 0·87
Lobular inflammation score (0-3) 1·7 (0·6) 1·6 (0·6) 1·7 (0·7) 0·82
Hepatocellular ballooning score (0-2) 0·8 (0·9) 0·7 (0·7) 1·1 (0·9) 0·14
Fibrosis stage (0-4) 1·3 (1·0) 1·2 (1·0) 1·1 (0·9) 0·70
1 Pediatric patients are from the TONIC trial with paired baseline and end of treatment biopsies and DNA. Total number of patients is 93.
Data are No. (%) or mean (SD).
2 P-value calculated using Fisher’s exact test for categorical characteristics and ANOVA for continuous characteristics.
3 6 patients refused to report a race; however, all 6 identified as Hispanic.
Abbreviations: BMI, body mass index; HDL, high density lipoprotein; HOMA-IR, homeostasis model assessment-estimated insulin resistance; Hp, haptoglobin; NAFLD, nonalcoholic fatty liver disease
Banini et al Haptoglobin and Vitamin E 3
Supplementary Table 3. Changes in histological features and liver enzymes after 96 weeks of treatment in adult patients by Haptoglobin genotype comparing Vitamin E and Placebo groups
Haptoglobin genotype No. improved/total No. (%) or
mean change from baseline ± SD OR or mean change VitE vs
placebo
1p
EvPl1
Interaction P
2VitE Placebo Hp Trend
Histological Outcomes Histologic improvement
32-2 14/29 (48%) 7/22 (32%) 2·0 0·24
n/c
4n/c
42-1 16/30 (53%) 10/38 (26%) 3·2 0·02
1-1 4/10 (40%) 0/6 (0%) 4·2 0·23
6Resolution of NASH
52-2 10/27 (37%) 2/17 (12%) 4·4 0·08
0·65 0.40
2-1 11/28 (39%) 6/35 (17%) 3·1 0·05
1-1 2/10 (20%) 1/5 (20%) 1·0 1·00
NAFLD Activity Score
62-2 -2·1 ± 1·8 -1·1 ± 1·8 -0·8 0·03
0·95 n/a
2-1 -1·0 ± 1·0 -0·2 ± 0·8 -1·4 0·001
1-1 -0·8 ± 1·8 1·2 ± 2·1 -0·4 0·25
Fibrosis stage
62-2 -0·4 ± 0·9 -0·3 ± 0·9 -0·2 0·40
2-1 -0·3 ± 1·2 -0·1 ± 0·9 -0·2 0·51 0·85 n/a
1-1 -0·8 ± 1·0 -0·2 ± 0·8 -0·4 0·45
Liver enzymes (U/L)
6Alanine aminotransferase
2-2 -42·8 ± 50·6 -17·2 ± 40·4 -25·2 <0·001
2-1 -31·6 ± 41·4 -20·1 ± 43·2 -13·1 0·04 0·28 0·08
1-1 -21·4 ± 76·3 -9·5 ± 31·1 -7·0 0·85
Aspartate aminotransferase
2-2 -27·4 ± 39·2 2·0 ± 19·9 -18·2 0·003
2-1 -21·0 ± 33·2 -8·2 ± 35·7 -14·8 0·02 0·25 0·13
1-1 -6·7 ± 25·3 -11·9 ± 39·1 5·8 0·74
Alkaline phosphatase
2-2 -10·9 ± 15·6 -0·2 ± 22·0 -11·7 0·02
2-1 -7·3 ± 14·5 -5·4 ± 14·4 -4·4 0·18 0·42 n/a
1-1 -4·6 ± 22·1 -4·3 ± 10·0 -6·2 0·51
γ-glutamyl transpeptidase
2-2 -19·8 ± 28·0 -1·6 ± 22·9 -20·2 0·003
2-1 -7·4 ± 28·5 -1·8 ± 32·0 -7·0 0·35 0·49 n/a
1-1 -13·0 ± 31·7 7·0 ± 18·4 -11·1 0·46
1 Odds ratios (OR) and P-values were calculated using logistic regression, regressing the binary outcome (improvement, resolution) on the treatment group for each haptoglobin genotype; p-values and mean changes from baseline were calculated using ANCOVA, regressing change from baseline to 96 weeks on treatment group and baseline value of the outcome for each haptoglobin genotype.
2 Interaction P-values calculated using logistic regression models for binary outcomes and linear regression models for continuous outcomes, regressing haptoglobin genotype (3-category) on treatment group by haptoglobin genotype, assuming the genotypic
association model. In calculating the P-values for the additive association model (trend), haptoglobin genotype was treated as an ordinal variable where Hp 1-1=0, Hp 2-1=1, and Hp 2-2=2. “n/a” denotes that the additive model was not appropriate due to violating model assumptions.
3 Histologic improvement defined as a decrease of at least 2 points in the NAS score and no worsening of fibrosis from baseline to end of treatment
4 Exact logistic was used to compute the OR and P-value due to small numbers in the treatment*Hp 1-1 group. “n/c” denotes that the interaction P-values could not be calculated due to small numbers.
5 For resolution of NASH, 13 patients with NAFLD but not NASH on the baseline biopsy were excluded from the analysis
6 Total N=135; Hp 2-2 vitamin E, n=29; Hp 2-2 placebo, n=22; Hp 2-1 vitamin E, n=30; Hp 2-1 placebo n=38; Hp 1-1 vitamin E, n=10;
Hp 1-1 placebo, n=6
Banini et al Haptoglobin and Vitamin E 4
Supplementary Table 4. Changes in histological features and liver enzymes after 96 weeks of treatment in pediatric patients by haptoglobin genotype comparing Vitamin E and Placebo groups Haptoglobin genotype No. improved/total No. (%) or mean
change from baseline ± SD OR or mean change VitE vs
placebo
1p
EvPl1Interaction P
2VitE Placebo Hp Trend Histological Outcomes
Histologic improvement
32-2 8/14 (57%) 0/13 (0%) 19·4 0·003
6n/c
4n/c
42-1 7/23 (30%) 7/21 (33%) 0·9 0·84
1-1 4/10 (40%) 3/11 (27%) 1·8 0·54
Resolution of NASH
52-2 8/14 (57%) 1/9 (11%) 10·7 0·05
0·59 0.45
2-1 12/19 (63%) 7/19 (37%) 2·9 0·10
1-1 3/8 (38%) 2/10 (20%) 2·4 0·42
NAFLD Activity Score
62-2 -2·5 ± 1·6 0·2 ± 1·2 -2·0 0·004
0·29 0·12
2-1 -1·3 ± 2·5 -0·7 ± 2·1 -0·7 0·24
1-1 -1·2 ± 1·3 -1·4 ± 2·1 0·5 0·95
Fibrosis stage
62-2 -0·5 ± 1·2 -0·1 ± 0·9 -0·4 0·27
2-1 -0·3 ± 0·9 -0·3 ± 1·4 0·0 0·96 0·65 n/a
1-1 0·2 ± 1·2 0·4 ± 1·4 -0·3 0·61
Liver enzymes (U/L)
6Alanine aminotransferase
2-2 -72·8 ± 74·9 -7·4 ± 31·7 -24·1 0·02
2-1 -48·7 ± 65·8 -20·6 ± 37·6 -22·5 0·22 0·11 0·04
1-1 -6·3 ± 76·3 -30·9 ± 49·1 31·5 0·22
Aspartate aminotransferase
2-2 -38·9 ± 46·5 -1·5 ± 24·9 -20·6 <0·001
2-1 -22·3 ± 33·2 -12·6 ± 36·6 -12·1 0·03 0·003 0·002
1-1 2·6 ± 23·8 -22·4 ± 43·0 16·0 0·13
Alkaline phosphatase
2-2 -67·6 ± 65·7 -32·3 ± 62·7 -24·5 0·26
2-1 -50·2 ± 78·9 -56·8 ± 59·9 -1·7 0·94 0·10 0·04
1-1 -20·4 ± 54·1 -61·9 ± 57·9 47·4 0·07
γ-glutamyl transpeptidase
2-2 -11·6 ± 17·4 7·8 ± 16·8 -15·9 0·05
2-1 -7·6 ± 19·5 -10·0 ± 29·7 -2·5 0·35 0·15 0·06
1-1 5·9 ± 20·4 -6·5 ± 23·5 11·6 0·27
1 Odds ratios (OR) and p-values were calculated using logistic regression, regressing the binary outcome (improvement, resolution) on the treatment group for each haptoglobin genotype; p-values and mean changes from baseline were calculated using ANCOVA, regressing change from baseline to 96 weeks on treatment group and baseline value of the outcome for each haptoglobin genotype·
2 Interaction P-values calculated using logistic regression models for binary outcomes and linear regression models for continuous
outcomes, regressing haptoglobin genotype (3-categorgy) on treatment group, assuming the genotypic association model· In calculating the P-values for the additive association model (trend), haptoglobin genotype was treated as an ordinal variable where Hp 1-1=0, Hp 2-1=1, and Hp 2-2=2· “n/a” denotes that the additive model was not appropriate due to violating model assumptions.
3 Histologic improvement defined as a decrease of at least 2 points in the NAS score and no worsening of fibrosis from baseline to end of treatment
4 Exact regression was used to compute the OR and P-value due to small numbers in the treatment*Hp 2-2 group. “n/c” denotes that the interaction P-values could not be calculated due to small numbers.
5 For resolution of NASH, 14 patients with NAFLD but not NASH on the baseline biopsy were excluded.
6 Total N=93; Hp 2-2 vitamin E, n=14; Hp 2-2 placebo, n=13; Hp 2-1 vitamin E, n=24; Hp 2-1 placebo n=21; Hp 1-1 vitamin E, n=10; Hp 1-1 placebo, n=11
