Supplemental Figure 1
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Supplemental Figure 1 Auranofin BSO Zn-BG A B C D carfilzomib
description
Supplemental Figure 1. Zn-BG. A. B. BSO. C. Auranofin. D. carfilzomib. Supplemental Table1: Characteristics of CLL patients in this study; * status unknown. Supplemental Figure 2. Supplemental Figure 3. A. B. Control. 1.0 m M AF, 24 h. CLL, p=.005. 150. NLC, p=ns. CLL-7607. - PowerPoint PPT Presentation
Transcript of Supplemental Figure 1
Supplemental Figure 1
Auranofin BSO Zn-BGA B C
D carfilzomib
Supplemental Table1: Characteristics of CLL patients in this study; * status unknown
Study labeling Gender IGHV ZAP 70 CD38 Cytogenetic alterations AF IC50 at 24h (M)
CLL 1 F M + + 11q-,13q- 0.24 CLL 2 F M - - 13q- 0.5 CLL 3 M U * * 17p- 0.22 CLL 4 M M * - +12 0.26 CLL 5 M U + - +12 0.15 CLL 6 M U * + * 0.2 CLL 7 M M - - 11q-, 13q-, 17p- 0.41 CLL 8 F M - - 13q- 1.75 CLL 9 F U * - 13q- 0.39 CLL 10 M * * - +12, 13q- 0.8 CLL 11 F U * - 13q- 0.34 CLL 12 M U * + 11q-, 13q- 0.27 CLL 13 M M * - 17p- 0.54 CLL 14 F M - - 13q- 0.24 CLL 15 M U * - 13q- 0.67 CLL 16 F U * - 11q-, 13q- 0.46 CLL 17 M * * + 13q- 1.33 CLL 18 M U - + 11q-, 13q- 0.34 CLL 19 M U * - 11q-, 13q- 0.53 CLL 20 F * + + 13q- 1.26 CLL 21 F M + - 17p- 0.73 CLL 22 M M - - 13q- 0.98 CLL 23 F U * + 17p- 0.66 CLL 24 M M - - 13q- 1.97 CLL 25 M U * + N 0.88 CLL 26 F U + + 17p- 0.16 CLL 27 M U - - 11q- 0.29 CLL 28 M U + + 11q- 0.18 CLL 29 F U - - 17p- 2.1 CLL 30 M U * - 11q-, +12 0.39 CLL 31 M M + - 11q-, 13q-, 17p- 0.69 CLL 32 F U + + 13q-, 17p- 0.36 CLL 33 M U + + 11q-, 13q- 0.72 CLL 34 F * * + 11q-, 13q- 1 CLL 35 F * * * 13q-, 17p- 1.92 CLL 36 M M - - 11q-, 13q- 0.68 CLL 37 F U * + +12 0.97 CLL 38 M U + 17p- 1.76 CLL 39 M * - - 11q-, 13q- 0.78 CLL 40 F * - - N 0.43 CLL 41 M * + + 13q-, 17p- 0.37 CLL 42 M * - - 17p- 0.72 CLL 43 M * * - 13q-, 17p- 0.67 CLL 44 M M - + +12 0.99 CLL 45 M M - + 11q-, 13q- 1.88 CLL 46 F U - - 17p- 0.88 CLL 47 F * * + +12, 17p- 1.16 CLL 48 F * + - +12 0.85 CLL 49 M U * - 13q-, 17p- 1.47 CLL 50 M U + + 13q- 0.78
Supplemental Figure 2
Control
100 nM, AF
250 nM, AF
500 nM, AF
1000 nM, AF
0
20
40
60
80
100
120 MEC-1
Control+ 2.5 mM NAC
% a
po
pto
sis,
48
hr
CLL
-760
7C
LL-7
620
Control 1.0 mM AF, 24 hA B
C
Supplemental Figure 3
NLC CLL
p=ns p=.005
-1.0
-0.8
-0.6
-0.4
-0.2
0.0
0.2
Re l
ativ
e c
ha
ng
e in
via
b ilit
y 1.0 mM AF, 24 h
0
50
100
150
Via
bili
ty (
%)
CLL, p=.005NLC, p=ns
1.0 m
M
AFContro
l
Before treatment After treatment
Mouse no. 1 6.96 x 106 1.31 x 106
Mouse no. 2 5.69x 106 9.00 x 105
Mouse no. 3 4.46 x 106 1.96 x 106
Mouse no. 4 8.73 x 106 2.32 x 106
Mouse no. 5 6.35 x 106 1.35 x 106
Leukemia cell burden
Supplemental Table 2: Leukemia cell burden for individual TCL-1 mice pre- and post-treatment with 2 weeks of auranofin.
Affy_ID Gene Fold Expression Change203665_at HMOX1 59.076203925_at GCLM 46.992225061_at DNAJA4 29.818
201841_s_at HSPB1 26.199208864_s_at TXN 18.697
36711_at MAFF 16.865211968_s_at HSP90AA1 11.342203810_at DNAJB4 9.474
203693_s_at E2F3 6.613201471_s_at SQSTM1 6.435207574_s_at GADD45B 5.875217144_at UBB 5.228
214359_s_at HSP90AB1 5.208210519_s_at NQO1 5.098204970_s_at MAFG 5.00225609_at GSR 4.996208680_at PRDX1 4.746
202284_s_at CDKN1A 4.267202417_at KEAP1 3.76236213_at NFE2L2 3.585200642_at SOD1 3.526210176_at TLR1 3.171236561_at TGFBR1 2.76218284_at SMAD3 2.532204194_at BACH1 2.332243423_at TNIP1 2.324206674_at FLT3 -2.324226068_at SYK -2.946225973_at TAP2 -2.579226117_at TIFA -2.808203508_at TNFRSF1B -2.58338290_at RGS14 -3.583206341_at IL2RA -2.656
205698_s_at MAP2K6 -4.603214130_s_at PDE4DIP -3.992214084_x_at NCF1C -3.134214259_s_at AKR7A2 -4.913204279_at PSMB9 -5.552
209040_s_at PSMB8 -4.022
Supplemental Table 3: Gene expression changes of the 39 most up and downregulated genes in primary CLL cells following treatment with 1.0 µM of AF for 8 hours
Network ID
Associated Network Functions Score
1 Protein Degradation, Protein Synthesis, Cellular Movement 41
2 Cellular Compromise, Cellular Function and Maintenance, Drug Metabolism 40
3 Free Radical Scavenging, Cellular Compromise, Molecular Transport 34
4 Endocrine System Disorders, Gastrointestinal Disease, Immunological Disease 29
5 Lipid Metabolism, Small Molecule Biochemistry, Infectious Disease 28
Supplemental Table 4: Ingenuity pathway analysis of the top 5 perturbed networks in Primary CLL cells treated with 1 µM of AF
Supplemental Figure 4
Zn-BG (µM) + AF (nM) FA CI
10 + 100 0.455 0.726
10 + 150 0.529 0.933
15 + 100 0.424 0.775
15 + 150 0.6215 0.765
20 + 100 0.4575 0.722
20 + 150 0.6155 0.775
MEC-1A.
Zn-BG (µM) + AF (nM) FA CI
10 + 100 0.472 0.803
10 + 250 0.625 0.520
10 + 500 0.806 0.144
15 + 100 0.510 0.578
15 + 250 0.675 0.323
15 + 500 0.826 0.107
20 + 250 .758 .132
20 + 500 0.815 0.128
Primary CLLB.
Supplemental Figure 5
C. D.
BSO (µM) + AF (nM) FA CI
1.0 + 10 0.0765
0.688
5.0 + 50 0.2435
0.443
7.5 + 75 0.6375
0.094
10 + 100 0.805 0.047
MEC-1
BSO (µM) + AF (nM) FA CI
1.0 + 250 0.58 0.547
2.0 + 250 0.558 0.761
3.0 + 250 0.650 0.463
4.0 + 250 0.820 0.201
5.0 + 250 0.80 0.239
Primary CLL
Supplemental Figure 6
β-Actin
CL. CASP3
MEC1
--
+-
-+
++ 2.5 mM NAC, 8 h
500 µM H2O2, 8 h
GRP78
Nrf2
CHOP
A
