ABCMdb

PMID: 23891399

Van Goor F, Yu H, Burton B, Hoffman BJ
Effect of ivacaftor on CFTR forms with missense mutations associated with defects in protein processing or function.
J Cyst Fibros. 2014 Jan;13(1):29-36. doi: 10.1016/j.jcf.2013.06.008. Epub 2013 Jul 23., [PubMed]

Sentences

No. Mutations Sentence Comment

0 ABCC7 p.Gly551Asp Original Article Effect of ivacaftor on CFTR forms with missense mutations associated with defects in protein processing or functionਮ,ਮਮ Fredrick Van Goor a,b,Ìe;, Haihui Yu b , Bill Burton b , Beth J. Hoffman b a Vertex Pharmaceuticals Incorporated, 130 Waverly St, Cambridge, MA, United States b Vertex Pharmaceuticals Incorporated, 11010 Torreyana Road, San Diego, CA 92121, United States Received 11 April 2013; received in revised form 6 June 2013; accepted 10 June 2013 Available online 23 July 2013 Abstract Background: Ivacaftor (KALYDECOࡊ, VX-770) is a CFTR potentiator that increased CFTR channel activity and improved lung function in patients age 6 years and older with CF who have the G551D-CFTR gating mutation. Login to comment 22 ABCC7 p.Trp1282* http://dx.doi.org/10.1016/j.jcf.2013.06.008 surface can also be due to CFTR mutations that either prevent the synthesis of full-length CFTR (e.g., W1282X: class I mutation) or reduce the amount of CFTR synthesis (e.g., 2789+5G࢐A: class V mutation). Login to comment 23 ABCC7 p.Gly551Asp ABCC7 p.Arg334Trp Other CFTR mutations do not affect the quantity of CFTR at the cell surface, but cause defects in CFTR channel gating (e.g., G551D: class III mutation), or reduce the ability of chloride to pass through the open channel pore (defective channel conductance; e.g., R334W; class IV mutation) [9-12]. Login to comment 28 ABCC7 p.Gly551Asp ABCC7 p.Gly551Ser ABCC7 p.Gly1244Glu ABCC7 p.Gly1349Asp ABCC7 p.Ser1255Pro ABCC7 p.Ser1251Asn ABCC7 p.Ser549Arg ABCC7 p.Ser549Asn ABCC7 p.Gly970Arg ABCC7 p.Gly178Arg These include the most common CFTR gating mutation, G551D, as well as the G178R, S549N, S549R, G551S, G970R, G1244E, S1251N, S1255P, and G1349D mutations [12]. Login to comment 29 ABCC7 p.Gly551Asp In patients ages 6 years and older with CF who have the G551D-CFTR gating mutation, ivacaftor increased chloride transport through CFTR and improved clinical measures of pulmonary function and nutritional status [14]. Login to comment 39 ABCC7 p.Ala455Glu ABCC7 p.Pro67Leu ABCC7 p.Glu92Lys However, a significantly higher level (P b 0.05; ANOVA followed by Tukey's multiple comparisons test; n = 3-6) of mRNA expression was measured for P67L-, E92K-, and A455E-CFTR (Fig. 1). Login to comment 42 ABCC7 p.His1085Arg ABCC7 p.Arg1066His For example, severe processing mutations such as F508del-, R1066H-, and H1085R-CFTR exhibited a low level of mature CFTR (b5% normal). Login to comment 43 ABCC7 p.Ala455Glu ABCC7 p.Ala455Glu ABCC7 p.Pro67Leu ABCC7 p.Pro67Leu ABCC7 p.Glu56Lys ABCC7 p.Glu56Lys Mutations such as P67L-, E56K-, and A455E-CFTR exhibited intermediate levels of mature CFTR, which are consistent with less severe defects in CFTR processing (mature CFTR protein for P67L-, E56K-, and A455E-CFTR were 28.4&#b1; 6.8, 12.2&#b1;1.5, and 11.5&#b1;2.5% of normal, respectively). Login to comment 44 ABCC7 p.Arg117His ABCC7 p.Ala455Glu ABCC7 p.Arg334Trp ABCC7 p.Arg347Pro ABCC7 p.Arg347His ABCC7 p.Asn1303Lys ABCC7 p.Asp1270Asn ABCC7 p.Arg74Trp ABCC7 p.Arg352Gln ABCC7 p.Leu206Trp ABCC7 p.Val520Phe ABCC7 p.Gly85Glu ABCC7 p.Arg560Thr ABCC7 p.Pro67Leu ABCC7 p.Thr338Ile ABCC7 p.Leu1065Pro ABCC7 p.Arg560Ser ABCC7 p.Met1101Lys ABCC7 p.Arg1070Gln ABCC7 p.Ser1235Arg ABCC7 p.Arg1066Cys ABCC7 p.Ser945Leu ABCC7 p.Asp1152His ABCC7 p.Tyr569Asp ABCC7 p.His1085Arg ABCC7 p.Phe1052Val ABCC7 p.Arg668Cys ABCC7 p.Arg117Cys ABCC7 p.Leu997Phe ABCC7 p.Leu1077Pro ABCC7 p.Phe1074Leu ABCC7 p.Arg1070Trp ABCC7 p.Glu92Lys ABCC7 p.Lys1060Thr ABCC7 p.Arg1066His ABCC7 p.Ile336Lys ABCC7 p.Ser492Phe ABCC7 p.Ala1067Thr ABCC7 p.Glu193Lys ABCC7 p.Asp110His ABCC7 p.Ser977Phe ABCC7 p.Ala559Thr ABCC7 p.Asp110Glu ABCC7 p.Asp579Gly ABCC7 p.His1054Asp ABCC7 p.Ala46Asp ABCC7 p.Ala561Glu ABCC7 p.Arg1066Met ABCC7 p.Leu927Pro ABCC7 p.Leu467Pro ABCC7 p.Glu56Lys ABCC7 p.Ser341Pro None M1V A46D E56K P67L R74W G85E E92K D110E D110H R117C R117H E193K L206W R334W I336K T338I S341P R347H R347P R352Q A455E L467P S492F F508del V520F A559T R560S R560T A561E Y569D D579G R668C L927P S945L S977F L997F F1052V H1054D K1060T L1065P R1066C R1066H R1066M A1067T R1070Q R1070W F1074L L1077P H1085R M1101K D1152H S1235R D1270N N1303K 0 100 200 300 400 500 600 * * * CFTR Mutation mRNA (% Normal CFTR) Fig. 1. Login to comment 53 ABCC7 p.Arg117Cys However, the following four exceptions were noted. The estimated total protein level for R117C-CFTR (38 &#b1; 5% normal CFTR) was lower (P b 0.05; ANOVA followed by Tukey's least significant difference test; n = 12) compared with normal CFTR, suggesting that the baseline level of chloride transport may be underestimated. Login to comment 54 ABCC7 p.Arg352Gln ABCC7 p.Asp1152His ABCC7 p.Glu193Lys In contrast, the estimated total protein levels for E193K-CFTR (177 &#b1; 12% normal CFTR; n = 6), R352Q-CFTR (178 &#b1; 4% normal CFTR; n = 6), and D1152H-CFTR (256 &#b1; 16% normal CFTR; n = 9) were higher (P b 0.05; ANOVA followed by Tukey's least significant difference test) compared with normal CFTR, suggesting that the baseline chloride transport may be overestimated by ~1.8 to 2.6 fold for these three mutant CFTR forms. Login to comment 58 ABCC7 p.Gly551Asp The EC50 of ivacaftor for all mutant CFTR forms tested was similar to G551D-CFTR (range; 101 to 735 nM) (Table 2; Fig. 3). Login to comment 64 ABCC7 p.Arg117His ABCC7 p.Ala455Glu ABCC7 p.Arg334Trp ABCC7 p.Arg347Pro ABCC7 p.Arg347His ABCC7 p.Asn1303Lys ABCC7 p.Asp1270Asn ABCC7 p.Arg74Trp ABCC7 p.Arg352Gln ABCC7 p.Leu206Trp ABCC7 p.Val520Phe ABCC7 p.Gly85Glu ABCC7 p.Arg560Thr ABCC7 p.Pro67Leu ABCC7 p.Thr338Ile ABCC7 p.Leu1065Pro ABCC7 p.Arg560Ser ABCC7 p.Met1101Lys ABCC7 p.Arg1070Gln ABCC7 p.Ser1235Arg ABCC7 p.Arg1066Cys ABCC7 p.Ser945Leu ABCC7 p.Asp1152His ABCC7 p.Tyr569Asp ABCC7 p.His1085Arg ABCC7 p.Phe1052Val ABCC7 p.Arg668Cys ABCC7 p.Arg117Cys ABCC7 p.Leu997Phe ABCC7 p.Leu1077Pro ABCC7 p.Phe1074Leu ABCC7 p.Arg1070Trp ABCC7 p.Glu92Lys ABCC7 p.Lys1060Thr ABCC7 p.Arg1066His ABCC7 p.Ile336Lys ABCC7 p.Ser492Phe ABCC7 p.Ala1067Thr ABCC7 p.Glu193Lys ABCC7 p.Asp110His ABCC7 p.Ser977Phe ABCC7 p.Ala559Thr ABCC7 p.Asp110Glu ABCC7 p.Asp579Gly ABCC7 p.His1054Asp ABCC7 p.Ala46Asp ABCC7 p.Ala561Glu ABCC7 p.Arg1066Met ABCC7 p.Leu927Pro ABCC7 p.Leu467Pro ABCC7 p.Glu56Lys ABCC7 p.Ser341Pro Mutant CFTR form CFTR processing Mature/total % Normal CFTR Normal 0.89 &#b1; 0.01 100.0 &#b1; 18.5 G85E -0.05 &#b1; 0.04 -1.0 &#b1; 0.9 R560S 0.00 &#b1; 0.00 0.0 &#b1; 0.0 R1066C 0.02 &#b1; 0.01 0.0 &#b1; 0.0 S492F 0.00 &#b1; 0.00 0.1 &#b1; 0.1 R560T 0.01 &#b1; 0.01 0.2 &#b1; 0.1 V520F 0.05 &#b1; 0.03 0.3 &#b1; 0.2 M1101K 0.05 &#b1; 0.03 0.3 &#b1; 0.1 A561E 0.08 &#b1; 0.04 0.5 &#b1; 0.2 R1066M 0.02 &#b1; 0.02 0.5 &#b1; 0.4 N1303K 0.02 &#b1; 0.02 0.5 &#b1; 0.3 A559T 0.16 &#b1; 0.09 0.6 &#b1; 0.2 M1V 0.06 &#b1; 0.06 0.7 &#b1; 0.6 Y569D 0.11 &#b1; 0.04 0.6 &#b1; 0.2 R1066H 0.08 &#b1; 0.02a 0.7 &#b1; 0.2a L1065P 0.05 &#b1; 0.05 1.0 &#b1; 0.8 L467P 0.10 &#b1; 0.07 1.2 &#b1; 0.8 L1077P 0.08 &#b1; 0.04 1.5 &#b1; 0.6 A46D 0.21 &#b1; 0.08 1.9 &#b1; 0.5a E92K 0.06 &#b1; 0.05 1.9 &#b1; 1.3 H1054D 0.09 &#b1; 0.04 1.9 &#b1; 0.8 F508del 0.09 &#b1; 0.02a 2.3 &#b1; 0.5a H1085R 0.06 &#b1; 0.01a 3.0 &#b1; 0.7a I336K 0.42 &#b1; 0.05a 6.5 &#b1; 0.7a L206W 0.35 &#b1; 0.10a 6.8 &#b1; 1.7a F1074L 0.52 &#b1; 0.03a 10.9 &#b1; 0.6a A455E 0.26 &#b1; 0.10a 11.5 &#b1; 2.5a E56K 0.29 &#b1; 0.04a 12.2 &#b1; 1.5a R347P 0.48 &#b1; 0.04a 14.6 &#b1; 1.8a R1070W 0.61 &#b1; 0.04a 16.3 &#b1; 0.6a P67L 0.36 &#b1; 0.04a 28.4 &#b1; 6.8a R1070Q 0.90 &#b1; 0.01a 29.5 &#b1; 1.4a S977F 0.97 &#b1; 0.01a 37.3 &#b1; 2.4a A1067T 0.78 &#b1; 0.03a 38.6 &#b1; 6.1a D579G 0.72 &#b1; 0.02a 39.3 &#b1; 3.1a D1270N 1.00 &#b1; 0.00a,c 40.7 &#b1; 1.2a S945L 0.65 &#b1; 0.04a 42.4 &#b1; 8.9a L927P 0.89 &#b1; 0.01a,b 43.5 &#b1; 2.5a,b R117C 0.87 &#b1; 0.02a,b 49.1 &#b1; 2.9a,b T338I 0.93 &#b1; 0.03a,b 54.2 &#b1; 3.7a,b L997F 0.90 &#b1; 0.04a,b 59.8 &#b1; 10.4a,b D110H 0.97 &#b1; 0.01a,b 60.6 &#b1; 1.5a,b S341P 0.79 &#b1; 0.02a 65.0 &#b1; 4.9a,b R668C 0.94 &#b1; 0.03a,b 68.5 &#b1; 1.9a,b R74W 0.78 &#b1; 0.01a 69.0 &#b1; 2.7a,b D110E 0.92 &#b1; 0.05a,b 87.5 &#b1; 9.5a,b R334W 0.91 &#b1; 0.05a,b 97.6 &#b1; 10.0a,b K1060T 0.87 &#b1; 0.02a,b 109.9 &#b1; 28.0a,b R347H 0.96 &#b1; 0.02a,c 120.7 &#b1; 2.8a,b S1235R 0.96 &#b1; 0.00a,c 139.0 &#b1; 9.0a,b E193K 0.84 &#b1; 0.02a,b 143.0 &#b1; 17.1a,b R117H 0.86 &#b1; 0.01a,b 164.5 &#b1; 34.2a,b R352Q 0.98 &#b1; 0.01a,b 179.9 &#b1; 8.0a,c F1052V 0.90 &#b1; 0.01a,b 189.9 &#b1; 33.1a,b D1152H 0.96 &#b1; 0.02a,c 312.0 &#b1; 45.5a,b Notes to Table 1: Quantification of steady-state CFTR maturation expressed as the mean (&#b1;SEM; n = 5-9) ratio of mature CFTR to total CFTR (immature plus mature) or level of mature mutant CFTR relative to mature normal-CFTR (% normal CFTR) in FRT cells individually expressing CFTR mutations. Login to comment 71 ABCC7 p.Ser1235Arg ABCC7 p.Asp1152His ABCC7 p.Phe1052Val ABCC7 p.Arg668Cys This was expected, as the CFTR mutations tested include known or putative CF-causing mutations, as well as CFTR mutations associated with varying clinical consequences (e.g., R668C, F1052V, D1152H) or complex CFTR alleles that may modify disease severity (e.g., S1235R) (www.CFTR2.org) [8,16]. Login to comment 74 ABCC7 p.Arg117His ABCC7 p.Arg117His ABCC7 p.Ala455Glu ABCC7 p.Ala455Glu ABCC7 p.Arg334Trp ABCC7 p.Arg334Trp ABCC7 p.Arg347Pro ABCC7 p.Arg347Pro ABCC7 p.Arg347His ABCC7 p.Arg347His ABCC7 p.Asn1303Lys ABCC7 p.Asn1303Lys ABCC7 p.Asp1270Asn ABCC7 p.Asp1270Asn ABCC7 p.Arg74Trp ABCC7 p.Arg74Trp ABCC7 p.Arg352Gln ABCC7 p.Arg352Gln ABCC7 p.Leu206Trp ABCC7 p.Leu206Trp ABCC7 p.Val520Phe ABCC7 p.Val520Phe ABCC7 p.Gly85Glu ABCC7 p.Gly85Glu ABCC7 p.Arg560Thr ABCC7 p.Arg560Thr ABCC7 p.Pro67Leu ABCC7 p.Pro67Leu ABCC7 p.Thr338Ile ABCC7 p.Thr338Ile ABCC7 p.Leu1065Pro ABCC7 p.Leu1065Pro ABCC7 p.Arg560Ser ABCC7 p.Arg560Ser ABCC7 p.Met1101Lys ABCC7 p.Met1101Lys ABCC7 p.Arg1070Gln ABCC7 p.Arg1070Gln ABCC7 p.Ser1235Arg ABCC7 p.Ser1235Arg ABCC7 p.Arg1066Cys ABCC7 p.Arg1066Cys ABCC7 p.Ser945Leu ABCC7 p.Ser945Leu ABCC7 p.Asp1152His ABCC7 p.Asp1152His ABCC7 p.Tyr569Asp ABCC7 p.Tyr569Asp ABCC7 p.His1085Arg ABCC7 p.His1085Arg ABCC7 p.Phe1052Val ABCC7 p.Phe1052Val ABCC7 p.Arg668Cys ABCC7 p.Arg668Cys ABCC7 p.Arg117Cys ABCC7 p.Arg117Cys ABCC7 p.Leu997Phe ABCC7 p.Leu997Phe ABCC7 p.Leu1077Pro ABCC7 p.Leu1077Pro ABCC7 p.Phe1074Leu ABCC7 p.Phe1074Leu ABCC7 p.Arg1070Trp ABCC7 p.Arg1070Trp ABCC7 p.Glu92Lys ABCC7 p.Glu92Lys ABCC7 p.Lys1060Thr ABCC7 p.Lys1060Thr ABCC7 p.Arg1066His ABCC7 p.Arg1066His ABCC7 p.Ile336Lys ABCC7 p.Ile336Lys ABCC7 p.Ser492Phe ABCC7 p.Ser492Phe ABCC7 p.Ala1067Thr ABCC7 p.Ala1067Thr ABCC7 p.Glu193Lys ABCC7 p.Glu193Lys ABCC7 p.Asp110His ABCC7 p.Asp110His ABCC7 p.Ser977Phe ABCC7 p.Ser977Phe ABCC7 p.Ala559Thr ABCC7 p.Ala559Thr ABCC7 p.Asp110Glu ABCC7 p.Asp110Glu ABCC7 p.Asp579Gly ABCC7 p.Asp579Gly ABCC7 p.His1054Asp ABCC7 p.His1054Asp ABCC7 p.Ala46Asp ABCC7 p.Ala46Asp ABCC7 p.Ala561Glu ABCC7 p.Ala561Glu ABCC7 p.Arg1066Met ABCC7 p.Arg1066Met ABCC7 p.Leu927Pro ABCC7 p.Leu927Pro ABCC7 p.Leu467Pro ABCC7 p.Leu467Pro ABCC7 p.Glu56Lys ABCC7 p.Glu56Lys ABCC7 p.Ser341Pro ABCC7 p.Ser341Pro Because the level of CFTR mRNA was similar across the panel of cell lines tested, the range in baseline activity and ivacaftor response likely reflects the severity of the functional defect and/or the 0 50 100 150 200 S341P R347P L467P S492F A559T A561E Y569D L1065P R1066C R1066M L1077P M1101K N1303K R560S L927P R560T H1085R V520F E92K M1V F508del H1054D I336K A46D G85E R334W T338I R1066H R352Q R117C L206W R347H S977F S945L A455E F1074L E56K P67L R1070W D110H D579G D110E R1070Q L997F A1067T E193K R117H R74W K1060T R668C D1270N D1152H S1235R F1052V Baseline With ivacaftor * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * Chloride transport (% Normal) Mutant CFTR form 0 100 200 300 400 S341P R347P L467P S492F A559T A561E Y569D L1065P R1066C R1066M L1077P M1101K N1303K R560S L927P R560T H1085R V520F E92K M1V F508del H1054D I336K A46D G85E R334W T338I R1066H R352Q R117C L206W R347H S977F S945L A455E F1074L P67L E56K R1070W D110H D579G D110E R1070Q L997F A1067T E193K R117H R74W K1060T R668C D1270N D1152H S1235R F1052V * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * * Mature CFTR (% Normal) Mutant CFTR form A B Fig. 2. Login to comment 82 ABCC7 p.Gly551Asp ABCC7 p.Arg117His ABCC7 p.Ala455Glu ABCC7 p.Arg334Trp ABCC7 p.Arg347Pro ABCC7 p.Arg347His ABCC7 p.Asn1303Lys ABCC7 p.Asp1270Asn ABCC7 p.Arg74Trp ABCC7 p.Arg352Gln ABCC7 p.Leu206Trp ABCC7 p.Val520Phe ABCC7 p.Gly85Glu ABCC7 p.Arg560Thr ABCC7 p.Pro67Leu ABCC7 p.Thr338Ile ABCC7 p.Leu1065Pro ABCC7 p.Arg560Ser ABCC7 p.Met1101Lys ABCC7 p.Arg1070Gln ABCC7 p.Ser1235Arg ABCC7 p.Arg1066Cys ABCC7 p.Ser945Leu ABCC7 p.Asp1152His ABCC7 p.Tyr569Asp ABCC7 p.His1085Arg ABCC7 p.Phe1052Val ABCC7 p.Arg668Cys ABCC7 p.Arg117Cys ABCC7 p.Leu997Phe ABCC7 p.Leu1077Pro ABCC7 p.Phe1074Leu ABCC7 p.Arg1070Trp ABCC7 p.Glu92Lys ABCC7 p.Lys1060Thr ABCC7 p.Arg1066His ABCC7 p.Ile336Lys ABCC7 p.Ser492Phe ABCC7 p.Ala1067Thr ABCC7 p.Glu193Lys ABCC7 p.Asp110His ABCC7 p.Ser977Phe ABCC7 p.Ala559Thr ABCC7 p.Asp110Glu ABCC7 p.Asp579Gly ABCC7 p.His1054Asp ABCC7 p.Ala46Asp ABCC7 p.Ala561Glu ABCC7 p.Arg1066Met ABCC7 p.Leu927Pro ABCC7 p.Leu467Pro ABCC7 p.Glu56Lys ABCC7 p.Ser341Pro Mutation Patientsa Chloride transport (bc;A/cm2 ) Chloride transport (% normal) EC50 Baseline With ivacaftor Baseline With ivacaftor Fold increase over baselineb Normal 204.5 &#b1; 33.3 301.3 &#b1; 33.8c 100.0 &#b1; 16.3 147.3 &#b1; 16.5c 1.5 266 &#b1; 42 G551D 1282 1.5 &#b1; 0.7 113.2 &#b1; 13.0c 1.0 &#b1; 0.5 55.3 &#b1; 6.3c 55.3 312 &#b1; 73 F1052V 12 177.3 &#b1; 13.7 410.2 &#b1; 11.3c 86.7 &#b1; 6.7 200.7 &#b1; 5.6c 2.3 177 &#b1; 14 S1235R ND 160.6 &#b1; 25.7 352.1 &#b1; 43.4c 78.5 &#b1; 12.6 172.2 &#b1; 21.2c 2.2 282 &#b1; 104 D1152H 185 117.3 &#b1; 23.0 282.7 &#b1; 46.9c 57.4 &#b1; 11.2 138.2 &#b1; 22.9c 2.4 178 &#b1; 67 D1270N 32 109.5 &#b1; 20.5 209.5 &#b1; 27.4c 53.6 &#b1; 10.0 102.4 &#b1; 13.4c 1.9 254 &#b1; 56 R668C 45 99.0 &#b1; 9.4 217.6 &#b1; 11.7c 48.4 &#b1; 4.6 106.4 &#b1; 5.7c 2.2 517 &#b1; 105 K1060T ND 89.0 &#b1; 9.8 236.4 &#b1; 20.3c 43.5 &#b1; 4.8 115.6 &#b1; 9.9c 2.7 131 &#b1; 73 R74W 25 86.8 &#b1; 26.9 199.1 &#b1; 16.8c 42.5 &#b1; 13.2 97.3 &#b1; 8.2c 2.3 162 &#b1; 17 R117H 739 67.2 &#b1; 13.3 274.1 &#b1; 32.2c 32.9 &#b1; 6.5 134.0 &#b1; 15.7c 4.1 151 &#b1; 14 E193K ND 62.2 &#b1; 9.8 379.1 &#b1; 1.1c 30.4 &#b1; 4.8 185.4 &#b1; 1.0c 6.1 240 &#b1; 20 A1067T ND 55.9 &#b1; 3.2 164.0 &#b1; 9.7c 27.3 &#b1; 1.6 80.2 &#b1; 4.7c 2.9 317 &#b1; 214 L997F 27 43.7 &#b1; 3.2 145.5 &#b1; 4.0c 21.4 &#b1; 1.6 71.2 &#b1; 2.0c 3.3 162 &#b1; 12 R1070Q 15 42.0 &#b1; 0.8 67.3 &#b1; 2.9c 20.6 &#b1; 0.4 32.9 &#b1; 1.4c 1.6 164 &#b1; 20 D110E ND 23.3 &#b1; 4.7 96.4 &#b1; 15.6c 11.4 &#b1; 2.3 47.1 &#b1; 7.6c 4.1 213 &#b1; 51 D579G 21 21.5 &#b1; 4.1 192.0 &#b1; 18.5c 10.5 &#b1; 2.0 93.9 &#b1; 9.0c 8.9 239 &#b1; 48 D110H 30 18.5 &#b1; 2.2 116.7 &#b1; 11.3c 9.1 &#b1; 1.1 57.1 &#b1; 5.5c 6.2 249 &#b1; 59 R1070W 13 16.6 &#b1; 2.6 102.1 &#b1; 3.1c 8.1 &#b1; 1.3 49.9 &#b1; 1.5c 6.2 158 &#b1; 48 P67L 53 16.0 &#b1; 6.7 88.7 &#b1; 15.7c 7.8 &#b1; 3.3 43.4 &#b1; 7.7c 5.6 195 &#b1; 40 E56K ND 15.8 &#b1; 3.1 63.6 &#b1; 4.4c 7.7 &#b1; 1.5 31.1 &#b1; 2.2c 4.0 123 &#b1; 33 F1074L ND 14.0 &#b1; 3.4 43.5 &#b1; 5.4c 6.9 &#b1; 1.6 21.3 &#b1; 2.6c 3.1 141 &#b1; 19 A455E 120 12.9 &#b1; 2.6 36.4 &#b1; 2.5c 6.3 &#b1; 1.2 17.8 &#b1; 1.2c 2.8 170 &#b1; 44 S945L 63 12.3 &#b1; 3.9 154.9 &#b1; 47.6c 6.0 &#b1; 1.9 75.8 &#b1; 23.3c 12.6 181 &#b1; 36 S977F 9 11.3 &#b1; 6.2 42.5 &#b1; 19.1c 5.5 &#b1; 3.0 20.8 &#b1; 9.3c 3.8 283 &#b1; 36 R347H 65 10.9 &#b1; 3.3 106.3 &#b1; 7.6c 5.3 &#b1; 1.6 52.0 &#b1; 3.7c 9.8 280 &#b1; 35 L206W 81 10.3 &#b1; 1.7 36.4 &#b1; 2.8c 5.0 &#b1; 0.8 17.8 &#b1; 1.4c 3.6 101 &#b1; 13 R117C 61 5.8 &#b1; 1.5 33.7 &#b1; 7.8c 2.9 &#b1; 0.7 16.5 &#b1; 3.8c 5.7 380 &#b1; 136 R352Q 46 5.5 &#b1; 1.0 84.5 &#b1; 7.8c 2.7 &#b1; 0.5 41.3 &#b1; 3.8c 15.2 287 &#b1; 75 R1066H 29 3.0 &#b1; 0.3 8.0 &#b1; 0.8c 1.5 &#b1; 0.1 3.9 &#b1; 0.4c 2.6 390 &#b1; 179 T338I 54 2.9 &#b1; 0.8 16.1 &#b1; 2.4c 1.4 &#b1; 0.4 7.9 &#b1; 1.2c 5.6 334 &#b1; 38 R334W 150 2.6 &#b1; 0.5 10.0 &#b1; 1.4c 1.3 &#b1; 0.2 4.9 &#b1; 0.7c 3.8 259 &#b1; 103 G85E 262 1.6 &#b1; 1.0 1.5 &#b1; 1.2 0.8 &#b1; 0.5 0.7 &#b1; 0.6 NS NS A46D ND 2.0 &#b1; 0.6 1.1 &#b1; 1.1 1.0 &#b1; 0.3 0.5 &#b1; 0.6 NS NS I336K 29 1.8 &#b1; 0.2 7.4 &#b1; 0.1c 0.9 &#b1; 0.1 3.6 &#b1; 0.1c 4 735 &#b1; 204 H1054D ND 1.7 &#b1; 0.3 8.7 &#b1; 0.3c 0.8 &#b1; 0.1 4.2 &#b1; 0.1c 5.3 187 &#b1; 20 F508del 29,018 0.8 &#b1; 0.6 12.1 &#b1; 1.7c 0.4 &#b1; 0.3 5.9 &#b1; 0.8c 14.8 129 &#b1; 38 M1V 9 0.7 &#b1; 1.4 6.5 &#b1; 1.9c 0.4 &#b1; 0.7 3.2 &#b1; 0.9c 8.0 183 &#b1; 85 E92K 14 0.6 &#b1; 0.2 4.3 &#b1; 0.8c 0.3 &#b1; 0.1 2.1 &#b1; 0.4c 7.0 198 &#b1; 46 V520F 58 0.4 &#b1; 0.2 0.5 &#b1; 0.2 0.2 &#b1; 0.1 0.2 &#b1; 0.1 NS NS H1085R ND 0.3 &#b1; 0.2 2.1 &#b1; 0.4 0.2 &#b1; 0.1 1.0 &#b1; 0.2 NS NS R560T 180 0.3 &#b1; 0.3 0.5 &#b1; 0.5 0.1 &#b1; 0.1 0.2 &#b1; 0.2 NS NS L927P 15 0.2 &#b1; 0.1 10.7 &#b1; 1.7c 0.1 &#b1; 0.1 5.2 &#b1; 0.8c 52.0 313 &#b1; 66 R560S ND 0.0 &#b1; 0.1 -0.2 &#b1; 0.2 0.0 &#b1; 0.0 -0.1 &#b1; 0.1 NS NS N1303K 1161 0.0 &#b1; 0.0 1.7 &#b1; 0.3 0.0 &#b1; 0.0 0.8 &#b1; 0.2 NS NS M1101K 79 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS L1077P 42 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS R1066M ND 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS R1066C 100 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS L1065P 25 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS Y569D 9 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS A561E ND 0.0 &#b1; 0.1 0.0 &#b1; 0.1 0.0 &#b1; 0.0 0.0 &#b1; 0.1 NS NS A559T 43 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS S492F 16 0.0 &#b1; 0.0 1.7 &#b1; 1.2 0.0 &#b1; 0.0 0.8 &#b1; 0.6 NS NS L467P 16 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS R347P 214 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 0.0 &#b1; 0.0 NS NS S341P 9 0.0 &#b1; 0.0 0.2 &#b1; 0.2 0.0 &#b1; 0.0 0.1 &#b1; 0.1 NS NS a Number of individuals with the individual mutation in the CFTR-2 database (www.CFTR2.org). Login to comment 86 ABCC7 p.Ala455Glu ABCC7 p.Asp1270Asn ABCC7 p.Leu206Trp ABCC7 p.Pro67Leu ABCC7 p.Arg1070Gln ABCC7 p.Ser945Leu ABCC7 p.Phe1074Leu ABCC7 p.Arg1070Trp ABCC7 p.Arg1066His ABCC7 p.Ala1067Thr ABCC7 p.Ser977Phe ABCC7 p.Asp579Gly ABCC7 p.His1054Asp ABCC7 p.Glu56Lys For example, the baseline level of chloride transport and ivacaftor response was higher for mutant CFTR forms associated with mild defects in CFTR processing (e.g., E56K, P67L, L206W, A455E, D579G, S945L, S977F, A1067T, R1070Q, R1070W, F1074L, and D1270N) than for those associated with severe defects in CFTR processing (e.g., F508del, H1054D, R1066H). Login to comment 87 ABCC7 p.Arg334Trp ABCC7 p.Arg347His ABCC7 p.Arg352Gln ABCC7 p.Thr338Ile ABCC7 p.Asp110His Similarly, the baseline chloride transport and ivacaftor response were higher for mutant CFTR forms with mild defects in channel conductance (30-84% of normal; D110H-, R347H, and R352Q-CFTR) [17-19], compared with those with severe defects in CFTR channel conductance (undetectable R334W- and T338I-CFTR) [9,20]. Login to comment 89 ABCC7 p.Arg117His ABCC7 p.Arg347Pro ABCC7 p.Arg1070Gln ABCC7 p.Ser945Leu ABCC7 p.Arg1070Trp ABCC7 p.Ala1067Thr For mutant CFTR forms that have multiple defects (e.g., R117H, F508del, S945L, R1070Q, A1067T, R1070W, and R347P), the relative impact of each defect is likely to affect the magnitude of the baseline chloride transport and ivacaftor response in vitro and in a clinical setting. Login to comment 90 ABCC7 p.Arg117His For example, the channel open probability and conductance of R117H-CFTR were reported to be 28% and 86% of normal CFTR, respectively [9]. Login to comment 91 ABCC7 p.Arg117His ABCC7 p.Arg347His ABCC7 p.Arg352Gln This suggests that the R117H mutation may be associated with defective channel gating, as well as conductance defects, and may explain why the ivacaftor response was larger than for CFTR mutations that result in defects in conductance alone (e.g., R347H, R352Q). Login to comment 92 ABCC7 p.Arg117His ABCC7 p.Ala455Glu ABCC7 p.Arg334Trp ABCC7 p.Arg334Trp ABCC7 p.Arg347His ABCC7 p.Asp1270Asn ABCC7 p.Arg74Trp ABCC7 p.Arg352Gln ABCC7 p.Leu206Trp ABCC7 p.Pro67Leu ABCC7 p.Thr338Ile ABCC7 p.Thr338Ile ABCC7 p.Arg1070Gln ABCC7 p.Ser1235Arg ABCC7 p.Ser945Leu ABCC7 p.Asp1152His ABCC7 p.Phe1052Val ABCC7 p.Arg668Cys ABCC7 p.Arg117Cys ABCC7 p.Leu997Phe ABCC7 p.Phe1074Leu ABCC7 p.Arg1070Trp ABCC7 p.Glu92Lys ABCC7 p.Glu92Lys ABCC7 p.Lys1060Thr ABCC7 p.Arg1066His ABCC7 p.Arg1066His ABCC7 p.Ala1067Thr ABCC7 p.Glu193Lys ABCC7 p.Asp110His ABCC7 p.Ser977Phe ABCC7 p.Asp110Glu ABCC7 p.Asp579Gly ABCC7 p.His1054Asp ABCC7 p.His1054Asp ABCC7 p.Glu56Lys Mutant CFTR forms that did not significantly respond to ivacaftor under the experimental conditions used in this study were generally associated with severe defects in CFTR processing A B C D E F 0 100 200 300 400 -9 -8 -7 -6 -5 -4 0 S1235R D1152H F1052V D1270N ivacaftor [Log M] 0 100 200 300 400 -9 -8 -7 -6 -5 -4 0 R668C K1060T R74W R117H ivacaftor [Log M] 0 100 200 300 400 -9 -8 -7 -6 -5 -4 0 E193K A1067T L997F R1070Q ivacaftor [Log M] Chloride Transport ( &#b5;A/cm 2 ) Chloride Transport ( &#b5;A/cm 2 ) Chloride Transport ( &#b5;A/cm 2 ) Chloride Transport ( &#b5;A/cm 2 ) Chloride Transport ( &#b5;A/cm 2 ) Chloride Transport ( &#b5;A/cm 2 ) Chloride Transport ( &#b5;A/cm 2 ) Chloride Transport ( &#b5;A/cm 2 ) Chloride Transport ( &#b5;A/cm 2 ) 0 100 200 300 400 -9 -8 -7 -6 -5 -4 0 D110E D579G D110H R1070W ivacaftor [Log M] 0 100 200 300 400 -9 -8 -7 -6 -5 -4 0 F1074L E56K P67L A455E ivacaftor [Log M] 0 100 200 300 400 -9 -8 -7 -6 -5 -4 0 R347H S945L L206W S977F ivacaftor [Log M] 0 100 200 300 400 -8 -6 -4 0 T338I R1066H R117C R352Q ivacaftor [Log M] 0 100 200 300 400 -9 -8 -7 -6 -5 -4 0 F508del R334W H1054D E92K ivacaftor [Log M] 0 5 10 15 20 -9 -8 -7 -6 -5 -4 0 F508del R334W H1054D E92K R1066H T338I ivacaftor [Log M] G H I Fig. 3. Login to comment 99 ABCC7 p.Ile336Lys ABCC7 p.His1054Asp A small but significant response to ivacaftor was also observed in FRT cells expressing the severe processing mutations, M1V, H1054D, and I336K (Table 2; Fig. 2). Login to comment 106 ABCC7 p.Pro67Leu For example, patients with CF who have the class II mutation, P67L, along with a severe CF-causing mutation (most commonly F508del), have a mean sweat chloride concentration of 57 mmol/L [21], which is lower than that in patients with CF who carry two copies of the most common class II mutation, F508del (~100 mmol/L). Login to comment 107 ABCC7 p.Arg117His ABCC7 p.Arg347Pro Similarly, patients with CF who have the most common class IV mutation, R117H, typically have a lower sweat chloride concentration than those with R347P (60 mmol/L vs. 99 mmol/L, respectively) [22]. Login to comment 113 ABCC7 p.Arg117His For example, the R117H CFTR mutation is relatively common among people heterozygous for the F508del CFTR mutation and is frequently identified in newborn screening programs [8,16]. Login to comment 114 ABCC7 p.Arg117His ABCC7 p.Arg117His When the R117H CFTR mutation is found on the same CFTR gene (in cis) with the 5T mutation (5 thymidine repeats in intron 8) a milder form of CF characterized by pancreatic sufficiency may be present, whereas when the 7T mutation in cis with R117H is found, CFTR-related disorders or no disease may be present. Login to comment 116 ABCC7 p.Asp1270Asn ABCC7 p.Arg74Trp ABCC7 p.Arg668Cys ABCC7 p.Gly576Ala ABCC7 p.Asp443Tyr Other examples of complex CFTR alleles include the number of TG repeats in intron 8 along with the 5T CFTR mutation (e.g., TG11-5T, TG12-5T, TG13-5T), R668C-G576A-D443Y, and R74W-D1270N [8,16]. Login to comment