Co-exposure to multiple metals, TERT-CLPTM1L variants, and their joint

influence on leukocyte telomere length

Yansen Bai1, Wenshan Fu1, Xin Guan, Xiulong Wu, Guyanan Li, Wei Wei, Yue Feng,

Hua Meng, Hang Li, Mengying Li, Ming Fu, Jiali Jie, Chenming Wang, Xiaomin

Zhang, Meian He, Huan Guo*

1 These authors contributed equally to this work.

Number of supplementary tables: 8.

Number of supplementary figures: 2.

1

Table S1. Components of PCR mixtures used for amplifying telomeric and β-globin sequences.

PCR mixtures

Telomeric

20 ng genomic DNA

1×master mix

90 nmol/L Tel1 primer: (5′-GGTTTTTGAGGGTGAGGGTGAGGGTGAGGGTGAGGGT-3′)

300 nmol/L Tel2 primer: (5′-TCCCGACTATCCCTATCCCTATCCCTATCCCTATCCCTA-3′)

β-globin

20 ng genomic DNA

1×master mix

150 nmol/L β-globin1 primer: (5′-GCTTCTGACACAACTGTGTTCACTAGC-3′)

150 nmol/L β-globin2 primer:(5′-CACCAACTTCATCCACGTTCACC-3′)

2

Table S2. The distributions of 23 plasma metals among our studied population.

Metals < LOQ, n (%)Percentiles (μg/L)

P5 P25 Median P75 P95

Aluminum (Al) 0 (0.0%) 20.01 34.46 52.34 92.53 210.17

Antimony (Sb) 0 (0.0%) 0.02 0.04 0.09 0.17 0.46

Arsenic (As) 2 (0.2%) 1.11 3.19 4.69 6.41 10.56

Barium (Ba) 0 (0.0%) 44.70 70.32 93.93 118.94 161.52

Cadmium (Cd) 23 (2.7%) 0.14 0.38 0.67 1.31 2.80

Chromium (Cr) 43 (5.1%) 0.36 1.27 1.73 2.51 5.87

Cobalt (Co) 191 (22.7%) 0.04 0.09 0.13 0.23 0.58

Copper (Cu) 2 (0.2%) 347.34 574.02 729.53 894.68 1213.54

Iron (Fe) 3 (0.4%) 521.58 908.80 1257.65 1664.69 2700.09

Lead (Pb) 6 (0.7%) 16.51 70.16 144.26 227.30 419.53

Manganese (Mn) 55 (6.5%) 0.66 1.97 2.73 4.21 10.68

Molybdenum (Mo) 0 (0.0%) 0.51 0.85 1.15 1.56 2.49

Nickel (Ni) 75 (8.9%) 0.40 1.78 2.69 4.82 18.15

Rubidium (Rb) 2 (0.2%) 255.98 348.45 401.41 465.99 601.86

Selenium (Se) 4 (0.5%) 21.63 47.44 66.75 84.21 114.41

Strontium (Sr) 2 (0.2%) 19.74 31.19 38.97 47.10 62.27

Thallium (Tl) 31 (3.7%) 0.06 0.10 0.12 0.17 0.31

Tin (Sn) 0 (0.0%) 0.07 0.22 0.22 0.68 4.57

Titanium (Ti) 3 (0.4%) 16.92 31.68 43.25 53.80 72.98

Tungsten (W) 0 (0.0%) 0.01 0.04 0.09 0.20 0.62

Uranium (U) 560 (66.5%) 0.02 0.02 0.02 0.04 0.19

Vanadium (V) 3 (0.4%) 0.33 0.55 0.73 1.00 1.83

Zinc (Zn) 0 (0.0%) 724.11 911.88 1108.30 1312.11 1715.78

Abbreviation: LOQ, limits of quantification.

3

Table S3. Pearson correlations between plasma metals.Metal

sAl As Ba Cd Co Cr Cu Fe Mn Mo Ni Pb Rb Sb Se Sn Sr Ti Tl V W Zn

Al1

-0.04

0.20* 0.06 0.48*

0.44*

0.35*

0.33*

0.78*

0.48* 0.61* -0.01

0.28* 0.43*

0.43* 0.40*

0.50*

0.52*

0.19*

0.75* 0.20*

0.29*

As1

0.74*

0.27*

-0.09*

0.12*

0.45*

0.36* -0.05

0.08*

-0.14*

0.53*

0.40*

-0.11*

0.47*

-0.26*

0.38*

0.38*

0.17*

0.07*

-0.19*

0.49*

Ba1

0.38* 0.07*

0.23*

0.72*

0.61*

0.17*

0.30* 0.12*

0.63*

0.73* -0.04

0.70*

-0.08*

0.65*

0.61*

0.17*

0.34*

-0.10*

0.79*

Cd1 0.08*

0.13*

0.24*

0.31*

0.09*

0.18* 0.18*

0.47*

0.28* 0.14*

0.33* 0.05

0.24*

0.29*

0.16*

0.20* 0.03

0.46*

Co1

0.47*

0.27*

0.17*

0.58*

0.53* 0.56* -0.04

0.16* 0.52*

0.35* 0.46*

0.33*

0.39*

0.43*

0.54* 0.37*

0.11*

Cr1

0.36*

0.33*

0.58*

0.35* 0.55*

0.09*

0.29* 0.26*

0.43* 0.33*

0.39*

0.45*

0.14*

0.49* 0.08*

0.28*

Cu1

0.67*

0.34*

0.49* 0.30*

0.35*

0.78* 0.05

0.84* 0.14*

0.79*

0.77*

0.22*

0.49* -0.02

0.68*

Fe1

0.37*

0.38* 0.31*

0.35*

0.71* 0.10*

0.67* 0.12*

0.62*

0.65*

0.21*

0.42* 0.01

0.59*

Mn1

0.51* 0.71* 0.03

0.28* 0.51*

0.44* 0.46*

0.48*

0.52*

0.25*

0.70* 0.21*

0.25*

Mo1 0.44*

0.12*

0.37* 0.37*

0.53* 0.35*

0.49*

0.55*

0.37*

0.60* 0.32*

0.35*

Ni1

0.11*

0.22* 0.52*

0.43* 0.52*

0.39*

0.48*

0.11*

0.60* 0.13*

0.30*

Pb1

0.37* -0.01

0.48*

-0.14*

0.33*

0.39* 0.02

0.17*

-0.12*

0.68*

Rb1 0.04

0.67* 0.07*

0.69*

0.56*

0.23*

0.43* 0.04

0.68*

Sb1

0.16* 0.49*

0.15*

0.19*

0.42*

0.45* 0.34*

0.04*

4

Se1 0.19*

0.76*

0.87*

0.19*

0.58* 0.005

0.72*

Sn1

0.19*

0.24*

0.21*

0.48* 0.28*

0.09*

Sr1

0.75*

0.22*

0.60* 0.04

0.64*

Ti1

0.21*

0.62* 0.03

0.62*

Tl1

0.26* 0.36* 0.05

V1 0.27*

0.53*

W 1 -0.07Zn 1

Abbreviation: Al, aluminum; As, arsenic; Ba, barium; Cd, cadmium; Co, cobalt; Cr, chromium; Cu, copper; Fe, iron; Mn, manganese; Mo, molybdenum; Ni, nickel; Pb, lead; Rb, rubidium; Sb, antimony; Se, selenium; Sn, tin; Sr, strontium; Ti, titanium; Tl, thallium; vanadium; W, tungsten; Zn, zinc.Notes: The concentrations of plasma metals were ln-transformed in Pearson correlation analyses.* The Pearson correlation coefficients were statistically significant (P < 0.05).

5

Table S4. The sensitivity analyses of plasma metals with leukocyte telomere length.Metals β 95%CIs PAluminum (Al) 0.013 -0.106, 0.132 0.830Antimony (Sb) 0.034 -0.031, 0.099 0.310Arsenic (As) -0.027 -0.109, 0.056 0.531Barium (Ba) -0.020 -0.132, 0.092 0.724Cadmium (Cd) 0.003 -0.061, 0.067 0.929Chromium (Cr) -0.001 -0.094, 0.092 0.986Cobalt (Co) -0.038 -0.123, 0.047 0.381Copper (Cu) -0.041 -0.151, 0.068 0.459Iron (Fe) 0.031 -0.061, 0.123 0.508Lead (Pb) -0.014 -0.060, 0.033 0.566Molybdenum (Mo) 0.005 -0.126, 0.136 0.939Nickel (Ni) -0.006 -0.085, 0.073 0.887Rubidium (Rb) 0.005 -0.101, 0.111 0.930Selenium (Se) -0.040 -0.139, 0.060 0.432Strontium (Sr) -0.097 -0.248, 0.055 0.211Thallium (Tl) 0.000 -0.105, 0.104 0.995Tin (Sn) 0.008 -0.039, 0.055 0.729Titanium (Ti) 0.006 -0.125, 0.138 0.927Tungsten (W) 0.018 -0.029, 0.065 0.463Vanadium (V) 0.016 -0.130, 0.162 0.833Zinc (Zn) -0.042 -0.139, 0.054 0.390Notes: The sensitivity analyses included ln-transformed LTL as dependent variable, ln-transformed plasma concentrations of metals as independent variables, adjusting for age, gender, BMI, smoking status (ever/never), alcohol drinking status (ever/never), levels of urinary ΣOH-PAHs, levels of plasma BPDE-Alb adducts and plasma manganese. 21 plasma metals were separately included in the model.

6

Table S5. Associations between quartile-categorized manganese and leukocyte telomere length.

Quartiles of manganese β (95%CIs)a P a

Q1 (<1.97 μg/L) Ref ---

Q2 (1.97 ~ 2.729 μg/L) 0.025 (-0.125, 0.175) 0.744Q3 (2.73 ~ 4.21 μg/L) 0.068 (-0.820, 0.218) 0.372Q4 (>4.21 μg/L) 0.230 (0.078, 0.383) 0.003Ptrend --- 0.003

a The model included ln-transformed LTL as the dependent variable, adjusting for age, gender, BMI, smoking status (ever/never), alcohol drinking status (ever/never), levels of urinary ΣOH-PAHs and plasma BPDE-Alb adducts.

7

Table S6. The 130 non-significant associations between TERT-CLPTM1L variants and leukocyte telomere length.

SNPs LocationMajor allele

Minor allele

MAF R2 GenotypedDominant model Recessive model Additive model

β STE Pdom β STE Prec β STE Padd

rs2853687 1256585 G A 0.054 0.87 Imputed 0.175 0.095 0.066 0.143 0.327 0.662 0.151 0.085 0.077rs35041195 1257288 C T 0.194 0.64 Imputed 0.050 0.060 0.409 0.066 0.140 0.636 0.043 0.050 0.391rs2736122 1257621 G A 0.054 0.99 Genotyped 0.144 0.095 0.130 0.143 0.327 0.662 0.126 0.086 0.140rs2736118 1260195 T C 0.057 0.82 Imputed 0.150 0.093 0.107 0.143 0.327 0.662 0.132 0.084 0.117rs11133715 1261052 G A 0.084 0.54 Imputed 0.023 0.078 0.766 0.276 0.357 0.439 0.032 0.074 0.661rs2736115 1264068 C T 0.056 0.81 Imputed 0.164 0.094 0.080 0.143 0.327 0.662 0.143 0.084 0.091rs2853685 1264152 A G 0.058 0.74 Imputed 0.134 0.092 0.145 0.143 0.327 0.662 0.119 0.083 0.152rs2736114 1265204 C T 0.057 0.75 Imputed 0.156 0.093 0.094 0.143 0.327 0.662 0.137 0.084 0.104rs2736113 1265373 G A 0.057 0.79 Imputed 0.156 0.093 0.094 0.143 0.327 0.662 0.137 0.084 0.104rs2736111 1265935 G A 0.057 0.78 Imputed 0.156 0.093 0.094 0.143 0.327 0.662 0.137 0.084 0.104rs2853684 1266226 T C 0.058 0.73 Imputed 0.156 0.093 0.094 0.030 0.302 0.922 0.125 0.083 0.130rs2075786 1266310 A G 0.150 0.63 Imputed 0.056 0.065 0.387 0.148 0.169 0.381 0.057 0.055 0.304rs3891054 1267202 C G 0.305 0.57 Imputed 0.073 0.057 0.204 0.028 0.097 0.774 0.048 0.044 0.273rs4246742 1267356 T A 0.316 0.58 Imputed 0.077 0.057 0.182 0.042 0.095 0.657 0.053 0.043 0.225rs11133719 1271524 C T 0.072 0.57 Imputed -0.048 0.083 0.564 -0.043 0.462 0.925 -0.045 0.080 0.569rs13172201 1271661 C T 0.154 0.58 Imputed -0.011 0.064 0.864 0.237 0.195 0.226 0.011 0.057 0.843rs78764532 1273215 G A 0.383 0.51 Imputed 0.015 0.060 0.801 0.047 0.084 0.576 0.020 0.043 0.641rs370474418 1273578 A G 0.168 0.53 Imputed -0.005 0.062 0.932 -0.141 0.196 0.472 -0.016 0.056 0.779rs562707895 1273610 T G 0.496 0.49 Imputed 0.026 0.066 0.691 -0.071 0.067 0.290 -0.017 0.041 0.688rs56081816 1273905 G A 0.065 0.47 Imputed -0.011 0.087 0.898 0.390 0.400 0.330 0.006 0.081 0.938rs4975605 1275528 C A 0.116 0.65 Imputed 0.053 0.070 0.454 0.325 0.234 0.167 0.066 0.063 0.295rs13156167 1275857 T C 0.229 0.68 Imputed 0.004 0.058 0.941 -0.003 0.126 0.982 0.003 0.048 0.958rs113136187 1276050 T C 0.166 0.80 Imputed -0.015 0.063 0.807 -0.052 0.181 0.776 -0.017 0.055 0.762rs556947195 1276772 A C 0.076 0.62 Imputed 0.076 0.081 0.348 0.054 0.400 0.893 0.070 0.077 0.360

8

rs55710479 1276785 C T 0.205 0.80 Imputed 0.019 0.059 0.746 0.013 0.159 0.937 0.016 0.052 0.758rs56000905 1276794 C A 0.208 0.80 Imputed 0.018 0.059 0.760 -0.008 0.154 0.957 0.013 0.051 0.806rs56345976 1276873 G A 0.251 0.99 Genotyped 0.051 0.058 0.375 0.058 0.124 0.640 0.043 0.047 0.363rs33961405 1277577 G A 0.432 0.68 Imputed 0.051 0.061 0.408 0.082 0.074 0.269 0.048 0.041 0.244rs2075785 1278584 C T 0.426 0.78 Imputed -0.005 0.062 0.942 -0.089 0.076 0.238 -0.029 0.042 0.482rs33963617 1279439 G A 0.051 0.66 Imputed 0.026 0.095 0.788 0.237 0.797 0.767 0.028 0.093 0.765rs10069690 1279790 C T 0.166 0.99 Genotyped -0.004 0.062 0.948 -0.301 0.190 0.114 -0.029 0.056 0.604rs10054203 1279964 G C 0.404 0.85 Imputed -0.027 0.060 0.653 -0.024 0.079 0.762 -0.020 0.042 0.635rs2242652 1280028 G A 0.163 1.00 Genotyped -0.020 0.062 0.749 -0.268 0.202 0.185 -0.037 0.056 0.512rs7734992 1280128 T C 0.398 0.87 Imputed -0.001 0.060 0.992 -0.028 0.081 0.725 -0.008 0.042 0.848rs13167280 1280477 G A 0.217 0.82 Imputed -0.004 0.059 0.945 0.052 0.131 0.695 0.004 0.049 0.929rs4975538 1280830 G C 0.389 0.86 Imputed -0.024 0.060 0.684 0.005 0.083 0.956 -0.011 0.043 0.793rs6897196 1280938 A G 0.390 0.89 Imputed -0.024 0.060 0.691 0.013 0.082 0.875 -0.009 0.043 0.839rs7726159 1282319 C A 0.411 1.00 Imputed -0.002 0.061 0.975 -0.050 0.080 0.530 -0.015 0.042 0.721rs7725218 1282414 G A 0.412 1.00 Genotyped -0.002 0.061 0.975 -0.042 0.079 0.598 -0.013 0.042 0.761rs7713218 1283312 G A 0.423 0.99 Imputed -0.002 0.061 0.968 -0.029 0.077 0.712 -0.010 0.042 0.819rs7717443 1283486 C T 0.423 0.99 Imputed -0.002 0.061 0.968 -0.029 0.077 0.712 -0.010 0.042 0.819rs72709458 1283755 C T 0.164 0.93 Imputed -0.010 0.063 0.878 -0.258 0.196 0.187 -0.029 0.056 0.608rs4449583 1284135 C T 0.412 0.99 Imputed 0.000 0.061 0.996 -0.050 0.080 0.530 -0.014 0.042 0.740rs80276247 1284717 C T 0.136 0.67 Imputed -0.056 0.065 0.388 -0.248 0.283 0.381 -0.061 0.061 0.317rs35029535 1284976 C T 0.086 0.74 Imputed 0.139 0.078 0.074 0.263 0.284 0.356 0.132 0.071 0.063rs10866498 1285162 T C 0.432 0.96 Imputed -0.011 0.062 0.865 -0.047 0.076 0.535 -0.019 0.042 0.646rs7705526 1285974 C A 0.415 1.00 Genotyped -0.009 0.061 0.888 -0.052 0.079 0.511 -0.019 0.042 0.651rs2736100 1286516 A C 0.412 1.00 Genotyped -0.017 0.061 0.784 -0.016 0.079 0.840 -0.013 0.042 0.766rs2853677 1287194 A G 0.385 1.00 Genotyped 0.039 0.059 0.510 -0.104 0.084 0.214 -0.007 0.043 0.873rs2736099 1287340 G A 0.439 0.91 Imputed -0.021 0.062 0.736 -0.037 0.074 0.616 -0.021 0.042 0.612rs7710703 1287505 C T 0.100 0.92 Imputed -0.038 0.073 0.603 0.177 0.302 0.559 -0.024 0.068 0.724

9

rs2853676 1288547 C T 0.163 0.99 Genotyped 0.023 0.062 0.709 0.018 0.208 0.933 0.020 0.056 0.718rs62332588 1289880 C T 0.405 0.81 Imputed -0.016 0.060 0.787 -0.004 0.081 0.963 -0.009 0.043 0.830rs74682426 1289975 C A 0.107 0.96 Imputed -0.010 0.071 0.890 0.330 0.283 0.243 0.010 0.066 0.885rs62332591 1290319 G T 0.453 0.86 Imputed -0.006 0.063 0.925 -0.047 0.073 0.523 -0.018 0.042 0.667rs536133213 1291124 T C 0.396 0.46 Imputed -0.056 0.060 0.351 -0.106 0.081 0.189 -0.057 0.042 0.179rs71575543 1291406 A G 0.332 0.57 Imputed -0.037 0.058 0.521 0.153 0.095 0.107 0.011 0.044 0.798rs71575544 1291418 G A 0.458 0.52 Imputed 0.053 0.063 0.401 0.114 0.071 0.108 0.061 0.041 0.140rs564256470 1291717 T C 0.380 0.51 Imputed -0.049 0.059 0.408 -0.095 0.082 0.242 -0.049 0.042 0.236rs528538268 1291718 G A 0.380 0.51 Imputed -0.049 0.059 0.408 -0.095 0.082 0.242 -0.049 0.042 0.236rs376986907 1291720 C T 0.380 0.51 Imputed -0.049 0.059 0.408 -0.095 0.082 0.242 -0.049 0.042 0.236rs56158232 1291735 A G 0.192 0.70 Imputed -0.020 0.060 0.738 -0.001 0.162 0.994 -0.015 0.052 0.769rs56023411 1291740 T G 0.191 0.70 Imputed -0.008 0.060 0.893 -0.001 0.162 0.994 -0.006 0.052 0.905rs2853672 1292983 A C 0.492 1.00 Genotyped -0.025 0.067 0.703 0.003 0.068 0.963 -0.009 0.042 0.833rs2736098 1294086 C T 0.378 1.00 Genotyped -0.038 0.059 0.518 0.043 0.083 0.608 -0.009 0.042 0.840rs2853669 1295349 A G 0.391 0.99 Genotyped -0.028 0.060 0.639 0.067 0.081 0.408 0.004 0.042 0.922rs7712562 1296072 G A 0.108 0.97 Imputed 0.001 0.071 0.988 0.330 0.283 0.243 0.019 0.066 0.775rs2735940 1296486 G A 0.487 1.00 Genotyped 0.001 0.066 0.983 0.004 0.069 0.959 0.002 0.042 0.965rs2736109 1296759 C T 0.327 1.00 Genotyped -0.014 0.057 0.805 0.053 0.092 0.563 0.004 0.043 0.932rs6554754 1297258 C T 0.108 0.96 Imputed 0.004 0.071 0.959 0.330 0.283 0.243 0.021 0.066 0.749rs2736108 1297488 C T 0.307 1.00 Genotyped 0.021 0.057 0.716 0.045 0.096 0.638 0.021 0.043 0.625rs2736107 1297854 C T 0.292 0.96 Imputed 0.012 0.057 0.838 0.064 0.099 0.522 0.019 0.044 0.660rs7449190 1297918 C T 0.123 0.92 Imputed 0.013 0.068 0.843 0.327 0.267 0.221 0.030 0.063 0.637rs3888705 1298645 G A 0.330 0.66 Imputed -0.060 0.058 0.299 -0.034 0.092 0.710 -0.041 0.043 0.342rs2736106 1298782 T C 0.439 0.66 Imputed -0.044 0.063 0.488 0.084 0.076 0.269 0.006 0.042 0.879rs62329683 1299087 C T 0.396 0.71 Imputed -0.096 0.059 0.104 -0.113 0.079 0.152 -0.078 0.041 0.060rs2735948 1299213 G A 0.170 0.77 Imputed 0.097 0.062 0.117 0.163 0.169 0.334 0.090 0.054 0.095rs2735846 1299379 C G 0.446 0.74 Imputed 0.081 0.062 0.191 -0.035 0.073 0.626 0.024 0.041 0.556

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rs2736105 1299756 T C 0.455 0.71 Imputed 0.101 0.063 0.108 -0.029 0.071 0.679 0.033 0.041 0.420rs13174814 1299859 G C 0.446 0.74 Imputed -0.026 0.063 0.682 -0.112 0.073 0.126 -0.048 0.042 0.253rs13174919 1299862 G C 0.448 0.74 Imputed -0.033 0.063 0.598 -0.123 0.073 0.090 -0.055 0.041 0.188rs2853668 1300025 G T 0.307 0.74 Imputed 0.048 0.057 0.402 0.078 0.101 0.439 0.044 0.044 0.321rs2853667 1300070 A G 0.369 0.73 Imputed 0.080 0.059 0.175 0.067 0.087 0.440 0.059 0.043 0.169rs4975612 1300310 G T 0.447 0.77 Imputed -0.016 0.063 0.803 -0.108 0.075 0.149 -0.042 0.043 0.323rs2736103 1300401 T C 0.462 0.76 Imputed 0.061 0.063 0.331 -0.040 0.070 0.567 0.012 0.041 0.770rs2735946 1300429 G T 0.088 0.73 Imputed 0.136 0.076 0.074 0.131 0.326 0.688 0.125 0.071 0.080rs11749061 1300439 G A 0.079 0.70 Imputed -0.118 0.079 0.138 0.142 0.400 0.723 -0.102 0.075 0.179rs2735845 1300584 C G 0.343 0.79 Imputed 0.043 0.058 0.453 0.018 0.090 0.840 0.028 0.043 0.513rs2736102 1302144 C T 0.160 0.85 Imputed 0.104 0.063 0.099 -0.002 0.176 0.991 0.079 0.055 0.151rs2853666 1302914 A G 0.163 0.86 Imputed 0.120 0.062 0.056 0.068 0.172 0.695 0.097 0.054 0.074rs2735945 1303901 C T 0.166 0.86 Imputed 0.098 0.062 0.115 0.071 0.169 0.674 0.081 0.054 0.133rs76236979 1304301 A C 0.068 0.82 Imputed 0.053 0.085 0.533 0.304 0.358 0.396 0.061 0.079 0.443rs4404721 1306165 C T 0.170 0.88 Imputed 0.111 0.062 0.073 0.033 0.162 0.838 0.085 0.053 0.108rs4530805 1306331 C T 0.171 0.89 Imputed 0.112 0.062 0.071 0.033 0.162 0.838 0.086 0.053 0.105rs7446461 1306521 G C 0.117 0.92 Imputed 0.077 0.070 0.271 0.030 0.201 0.883 0.061 0.061 0.316rs180725355 1307555 T C 0.124 0.83 Imputed 0.124 0.068 0.067 -0.047 0.243 0.848 0.101 0.062 0.104rs185381850 1307558 T G 0.124 0.83 Imputed 0.124 0.068 0.067 -0.047 0.243 0.848 0.101 0.062 0.104rs200878815 1307629 T C 0.132 0.84 Imputed 0.102 0.067 0.126 0.074 0.208 0.720 0.087 0.059 0.143rs201857317 1307632 G T 0.480 0.75 Imputed 0.012 0.064 0.852 -0.038 0.068 0.575 -0.009 0.040 0.828rs183257831 1307642 A G 0.452 0.74 Imputed 0.005 0.063 0.932 0.007 0.071 0.927 0.004 0.041 0.914rs62651531 1307910 A G 0.229 0.75 Imputed 0.034 0.058 0.553 0.078 0.133 0.559 0.034 0.049 0.478rs4635969 1308552 G A 0.120 1.00 Genotyped 0.077 0.069 0.269 0.034 0.195 0.862 0.061 0.060 0.312rs61574973 1309168 C T 0.173 0.97 Imputed 0.119 0.062 0.054 0.108 0.159 0.496 0.099 0.053 0.061rs60622800 1309904 A G 0.173 0.96 Imputed 0.119 0.062 0.054 0.108 0.159 0.496 0.099 0.053 0.061rs6554758 1310152 A G 0.173 0.96 Imputed 0.119 0.062 0.054 0.108 0.159 0.496 0.099 0.053 0.061

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rs12332579 1311198 T C 0.053 0.90 Imputed 0.177 0.093 0.058 -0.024 0.567 0.966 0.164 0.090 0.069rs6866294 1311693 C T 0.173 0.97 Imputed 0.119 0.062 0.054 0.108 0.159 0.496 0.099 0.053 0.061rs6866783 1312020 C T 0.173 0.97 Imputed 0.119 0.062 0.054 0.108 0.159 0.496 0.099 0.053 0.061rs35953391 1312329 C T 0.117 0.98 Imputed 0.077 0.070 0.271 0.030 0.201 0.883 0.061 0.061 0.316rs13356727 1312457 A G 0.174 0.97 Imputed 0.120 0.061 0.052 0.108 0.159 0.496 0.100 0.053 0.059rs13355267 1312935 C T 0.174 0.97 Imputed 0.120 0.061 0.052 0.108 0.159 0.496 0.100 0.053 0.059rs36115365 1313242 G C 0.117 0.98 Imputed 0.078 0.070 0.263 0.030 0.201 0.883 0.062 0.061 0.307rs28379291 1313701 G A 0.175 0.98 Imputed 0.118 0.061 0.055 0.108 0.159 0.496 0.099 0.053 0.061rs10078017 1314009 T C 0.175 0.98 Imputed 0.118 0.061 0.055 0.108 0.159 0.496 0.099 0.053 0.061rs4975615 1315343 A G 0.175 0.99 Imputed 0.118 0.061 0.055 0.108 0.159 0.496 0.099 0.053 0.061rs4975616 1315660 A G 0.175 1.00 Genotyped 0.118 0.061 0.055 0.108 0.159 0.496 0.099 0.053 0.061rs13170453 1317481 A G 0.117 0.97 Imputed 0.068 0.070 0.331 0.031 0.196 0.875 0.054 0.061 0.374rs3816659 1317820 G A 0.175 0.99 Imputed 0.117 0.061 0.057 0.108 0.159 0.496 0.098 0.053 0.063rs451360 1319680 C A 0.113 0.98 Imputed 0.053 0.071 0.458 0.082 0.208 0.692 0.048 0.062 0.442rs402710 1320722 C T 0.330 0.99 Genotyped 0.030 0.058 0.596 -0.120 0.092 0.192 -0.009 0.043 0.833rs401681 1322087 C T 0.339 1.00 Genotyped 0.061 0.058 0.296 -0.094 0.092 0.305 0.013 0.043 0.766rs428499 1322663 C T 0.339 1.00 Genotyped 0.058 0.058 0.313 -0.036 0.088 0.686 0.023 0.042 0.585rs410805 1323196 C T 0.336 0.98 Imputed 0.056 0.058 0.330 -0.062 0.089 0.484 0.016 0.042 0.701rs139395514 1328320 A G 0.158 0.82 Imputed 0.112 0.063 0.074 0.147 0.195 0.452 0.102 0.056 0.070rs380145 1328897 C T 0.113 0.97 Imputed 0.053 0.071 0.458 0.082 0.208 0.692 0.048 0.062 0.442rs56190840 1333778 A C 0.464 0.49 Imputed 0.012 0.063 0.843 0.018 0.068 0.796 0.011 0.040 0.783rs371277671 1333779 A T 0.464 0.49 Imputed 0.012 0.063 0.843 0.018 0.068 0.796 0.011 0.040 0.783rs397640 1334729 C T 0.074 0.88 Imputed 0.027 0.083 0.743 0.128 0.327 0.696 0.030 0.076 0.694rs74739441 1337174 C T 0.096 1.00 Genotyped -0.033 0.075 0.657 0.038 0.254 0.881 -0.024 0.067 0.718rs36019446 1339890 A G 0.185 0.88 Imputed 0.105 0.061 0.084 0.054 0.153 0.725 0.082 0.052 0.112Abbreviation: MAF, minor allele frequency; STE, standard error; dom, dominant; rec, recessive; add, additive.Notes: Dominant, recessive, and additive models were conducted to investigate the associations between TERT-CLPTM1L variants and ln-transformed TL in the generalized

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linear regression, with adjustment for age and gender. In the additive model, P values were calculated for the trend of β with an increased number of minor alleles.

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Table S7. The expression Quantitative Trait Loci analyses of 34 positive SNPs in TERT and CLPTMIL.

SNPs LocationMajor allele

Minor allele

TERT CLPTM1LEffect size

P TissuesEffect size

P Tissues

rs33954691 1255520 G A --- --- --- --- --- ---rs6554759 1317102 A G --- --- --- --- --- ---Block 1

rs1801075 1317949 T C --- --- --- --- --- ---rs421629 1320136 G A -0.260 5.70E-08 Esophagus – Mucosa -0.130 1.20E-07 Skinrs380286 1320247 G A -0.270 1.50E-08 Esophagus – Mucosa -0.130 1.70E-07 Skinrs10073340 1321873 C T --- --- --- --- --- ---rs381949 1322468 G A -0.260 1.70E-07 Esophagus – Mucosa -0.120 2.60E-06 Skinrs13178866 1323212 C T -0.270 1.30E-08 Esophagus – Mucosa -0.140 5.90E-08 Skinrs414965 1324121 G A -0.260 2.20E-07 Esophagus – Mucosa -0.120 6.60E-06 Skinrs421284 1325590 T C -0.270 5.00E-09 Esophagus – Mucosa -0.130 1.90E-07 Skinrs465498 1325803 A G -0.270 8.70E-09 Esophagus – Mucosa -0.130 3.20E-07 Skinrs11133729 1327101 C G --- --- --- --- --- ---rs383009 1327851 C T --- --- --- --- --- ---rs452932 1330253 T C -0.270 5.00E-09 Esophagus – Mucosa -0.130 1.70E-07 Skinrs452384 1330840 T C -0.280 3.90E-09 Esophagus – Mucosa -0.130 2.10E-07 Skinrs370348 1331219 A G -0.280 3.90E-09 Esophagus – Mucosa -0.130 2.10E-07 Skinrs62329699 1331685 T C --- --- --- --- --- ---rs2447853 1333077 A G -0.270 1.10E-08 Esophagus – Mucosa -0.140 8.90E-08 Skin

rs2455393 1333231 G T --- --- --- --- --- ---Block 2

rs457130 1336178 A T -0.250 2.10E-07 Esophagus - Mucosa -0.120 7.00E-06 Skin

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rs467095 1336221 T C -0.260 3.50E-08 Esophagus - Mucosa -0.130 2.20E-07 Skinrs455433 1336243 A G -0.270 5.00E-09 Esophagus - Mucosa -0.130 1.70E-07 Skinrs460073 1336459 T C -0.270 5.00E-09 Esophagus - Mucosa -0.130 1.70E-07 Skinrs462608 1336626 T A -0.260 1.30E-07 Esophagus - Mucosa -0.120 7.20E-06 Skinrs456366 1337070 T C -0.260 3.50E-08 Esophagus - Mucosa -0.130 2.20E-07 Skinrs459961 1337106 T A -0.270 5.00E-09 Esophagus - Mucosa -0.130 1.70E-07 Skinrs31484 1337906 A T -0.260 3.50E-08 Esophagus - Mucosa -0.130 2.20E-07 Skinrs71575564 1339797 T C --- --- --- --- --- ---rs42269 1339985 C A --- --- --- --- --- ---rs35595862 1340252 G A --- --- --- --- --- ---rs31487 1341101 G C -0.270 1.00E-08 Esophagus - Mucosa -0.130 2.00E-07 Skinrs31489 1342714 C A -0.250 2.30E-07 Esophagus - Mucosa -0.120 3.90E-06 Skinrs10462706 1343794 C T --- --- --- --- --- ---rs31490 1344458 G A -0.270 2.20E-08 Esophagus - Mucosa -0.140 4.90E-08 Skin

Notes: The above analyses were based on the online tool (GTEx, http://www.gtexportal.org/home/). The effect size above zero indicated the increased expression with an elevated number of minor alleles. The bolded SNPs were tagSNPs.

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Table S8. Association of plasma manganese level with leukocyte telomere length by different genotypes.

SNPs Genotypes

Leukocyte telomere length

β (95%CIs) P PinteractionLow plasma manganese

(≤4.21 μg/L)High plasma manganese

(>4.21 μg/L)n median (25th, 75th) n median (25th, 75th)

rs33954691 GG 390 1.86 (1.00, 2.85) 121 2.30 (1.59, 3.20) 0.273 (0.108, 0.438) 0.0010.172

GA+AA 196 1.76 (0.91, 2.63) 74 2.01 (1.17, 2.97) 0.105 (-0.113, 0.323) 0.347rs6554759 AA 521 1.81 (0.98, 2.82) 172 2.19 (1.38, 3.09) 0.205 (0.066, 0.344) 0.004

0.434AG+GG 65 1.76 (1.05, 3.56) 23 2.75 (2.00, 3.94) 0.342 (-0.070, 0.753) 0.104

rs465498 AA 402 1.81 (0.93, 2.83) 135 2.18 (1.34, 3.14) 0.200 (0.040, 0.360) 0.0140.665

AG+GG 184 1.83 (1.08, 2.85) 60 2.45 (1.68, 3.18) 0.223 (-0.005, 0.451) 0.056rs2455393 GG 323 1.80 (0.98, 2.69) 106 2.12 (1.27, 3.03) 0.161 (-0.021, 0.343) 0.083

0.326GT+TT 263 1.83 (0.99, 2.97) 89 2.42 (1.62, 3.20) 0.296 (0.108, 0.485) 0.002

rs31489 CC 401 1.81 (0.93, 2.83) 136 2.19 (1.34, 3.16) 0.210 (0.050, 0.369) 0.0100.807

CA+AA 185 1.85 (1.09, 2.80) 59 2.42 (1.67, 3.14) 0.213 (-0.015, 0.441) 0.068Notes: Generalized linear model with ln-transformed LTL as dependent variable, plasma levels of manganese (low/high) as independent variable was conducted, with adjustment for age, gender, BMI, smoking status (ever/never), alcohol drinking status (ever/never), levels of urinary ΣOH-PAHs and plasma BPDE-Alb adducts. Pinteraction was calculated by introducing a multiplicative interaction term (dichotomous plasma manganese × genotypes) into the generalized linear models. All subjects were divided into low and high manganese subgroups by the 75th percentile (4.21 μg/L) of plasma manganese level.

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.

Fig. S1. Association between plasma manganese level and leukocyte telomere length among different subgroups.

Notes: Generalized linear model was conducted, with ln-transformed LTL as the dependent variable, and ln-transformed plasma concentrations of manganese as independent variable, with adjustment for age, gender, BMI, smoking status (ever/never), alcohol drinking status (ever/never), levels of urinary ΣOH-PAHs and plasma BPDE-Alb adducts. Pinteraction was calculated by introducing a multiplicative interaction term (plasma manganese level × characteristics’ subgroups) into the generalized linear models. The red solid square and blue line in panel represent β (95% CI).

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Fig. S2. The linkage disequilibrium analysis for 34 LTL-related SNPs among our study participants. Notes: The values in the box equaled to the D'×100 between SNPs. The colors from white to red indicated the level of linkage disequilibrium

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