TABLE 3.
Susceptibility of HCV GT-6 NS5A polymorphisms at residue positions associated with NS5A drug resistance
| GT-6a NS5A rep bkgda | RASb | ECs (±SD)c [nM] for: |
|||||||
|---|---|---|---|---|---|---|---|---|---|
| DCV |
LDV |
OMV |
VEL |
||||||
| EC50 | EC90 | EC50 | EC90 | EC50 | EC90 | EC50 | EC90 | ||
| Cons | Cons | 0.30 (0.02) | 0.70 (0.04) | 4.3 (0.2) | 14 (1) | 8.1 (0.8) | 19 (1) | 0.05 (0.01) | 0.10 (0.01) |
| Cons | L28A | 296 (20) | 735 (41) | 677 (31) | 989 (142) | 1,720 (107) | 2,032 (131) | 100 (8) | 331 (24) |
| Cons | L28F | 0.65 (0.04) | 1.4 (0.1) | 38 (2) | 91 (6) | 13 (1) | 34 (3) | 0.09 (0.01) | 0.20 (0.03) |
| Cons | L28M | 9.3 (0.8) | 22 (1) | 152 (5) | 250 (22) | 21 (1) | 39 (2) | 0.60 (0.07) | 1.5 (0.2) |
| Cons | L28T | 224 (13) | 432 (32) | 560 (3) | 620 (60) | 1,483 (49) | 1,683 (21) | 54 (1) | 119 (4) |
| Cons | L28V | 1.9 (0.1) | 4.5 (0.1) | 31 (2) | 75 (4) | 81 (3) | 177 (11) | 0.55 (0.02) | 1.3 (0.1) |
| Cons | R30A | 0.40 (0.02) | 0.90 (0.02) | 1.5 (0.1) | 5.3 (0.2) | 0.0020 (0.0001) | 0.009 (0.001) | 0.0100 (0.0004) | 0.040 (0.002) |
| Cons | L31I | 6.4 (0.0) | 30 (1) | 334 (22) | 891 (141) | 108 (7) | 365 (15) | 1.0 (0.2) | 3.2 (0.3) |
| Cons | T93I | 1.2 (0.1) | 2.7 (0.1) | 21 (2) | 45 (1) | 29 (3) | 55 (12) | 0.08 (0.00) | 0.25 (0.02) |
| Cons | T93S | 1.1 (0.0) | 2.7 (0.2) | 25 (2) | 68 (5) | 46 (5) | 90 (7) | 0.15 (0.01) | 0.35 (0.02) |
| Cons | L28M, R30S | 47 (4) | 218 (22) | 188 (17) | 481 (33) | 0.25 (0.02) | 0.75 (0.06) | 0.30 (0.02) | 0.65 (0.20) |
| Cons | L28T, R30S | 735 (72) | 1,081 (101) | 876 (226) | 1,113 (279) | 1,746 (16) | 2,056 (16) | 63 (5) | 128 (39) |
| Cons | L28V, R30A | 0.55 (0.07) | 1.4 (0.2) | 1.8 (0.1) | 4.7 (0.2) | 0.085 (0.01) | 0.25 (0.01) | 0.05 (0.00) | 0.18 (0.01) |
| Cons | L28V, R30S | 8.7 (0.7) | 20 (2) | 62 (1) | 76 (2) | 9.5 (0.3) | 25 (1) | 0.25 (0.01) | 0.85 (0.01) |
| Cons | L28V, R30T | 4.8 (0.3) | 14 (1) | 38 (3) | 115 (3) | 2.9 (0.2) | 11 (0) | 0.09 (0.01) | 0.20 (0.02) |
| Cons | L28T, L31I | 2349 (128) | >5,000 | >5,000 | >5,000 | >5,000 | >5,000 | 42,221 (750) | >5,000 |
| Cons | L28V, R30S, T93S | 91 (13) | 254 (48) | 296 (31) | 618 (75) | 174 (27) | 418 (26) | 3.9 (0.4) | 12 (1) |
| JFH-1 | Cons | 0.10 (0.01) | 0.20 (0.01) | 2.7 (0.3) | 5.6 (0.1) | 5.2 (1.3) | 7.0 (0.2) | 0.006 (0.004) | 0.02 (0.01) |
| JFH-1 | L31M | 85 (3) | 137 (5) | 398 (97) | 544 (29) | 26 (2) | 43 (1) | 5.2 (0.5) | 8.3 (0.6) |
| JFH-1 | T58A | 3.6 (0.3) | 6.3 (0.2) | 33 (1) | 52 (1) | 74 (5) | 124 (19) | 0.023 (0.001) | 0.050 (0.003) |
| JFH-1 | T58S | 3.5 (0.1) | 6.1 (0.1) | 41 (13) | 56 (5) | 44 (8) | 70 (1) | 0.020 (0.000) | 0.05 (0.01) |
| Con1 | Cons | 0.088 (0.005) | 0.39 (0.07) | 0.60 (0.06) | 3.2 (0.2) | 1.4 (0.1) | 9.0 (1.0) | 0.014 (0.001) | 0.043 (0.005) |
| Con1 | R30E | 1.8 (0.0) | 9.6 (0.8) | 4.2 (0.8) | 29 (5) | 0.31 (0.05) | 1.6 (0.2) | 0.018 (0.001) | 0.075 (0.012) |
| Con1 | R30N | 0.35 (0.02) | 1.7 (0.1) | 0.53 (0.03) | 4.3 (0.2) | 0.003 (0.001) | 0.013 (0.002) | 0.009 (0.001) | 0.027 (0.002) |
| Con1 | R30S | 0.33 (0.03) | 1.6 (0.1) | 0.51 (0.04) | 3.7 (0.3) | 0.003 (0.001) | 0.012 (0.001) | 0.009 (0.001) | 0.024 (0.003) |
| Con1 | L28A R30S | 1,139 (155) | 3,269 (486) | 1,263 (43) | 2,648 (265) | 1,121 (123) | 1,630 (59) | 183 (60) | 720 (114) |
| Con1 | L28M, R30N | 20 (3) | 181 (20) | 74 (4) | 370 (27) | 0.024 (0.006) | 0.096 (0.016) | 0.61 (0.02) | 2.7 (0.1) |
a
GT-6a NS5A consensus (Cons) sequence was introduced into GT-2a (JFH-1) and GT-1b (Con1) replicon constructs in case GT-6a NS5A substitutions did not result in viable replicons in the GT-6a (NS3-5B) consensus replicon background (rep bkgd).
b
Certain GT-6 NS5A replicon constructs harboring various substitutions and combinations of substitutions did not replicate in our replicon system. NS5A-A92P conferred no change in NS5A drug potencies compared with that of the HCV GT-6a-cons replicon (data not shown).
c
EC, effective concentration; SD, standard deviation.