Introduction
Methods
Study design
-
Group 1: 155 patients with anatomic coronary alteration (comprising patients with acute coronary syndrome and chronic stable angina).
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Group 2: 46 patients with functional coronary alteration [normal coronary arteries as assessed by angiography, and microvascular dysfunction defined as coronary flow reserve (CFR) < 2.5 after IC infusion of acetylcholine and adenosine].
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Group 3: 41 patients with anatomically and functionally normal coronary arteries as assessed by angiography and with normal functional tests (CFR ≥ 2.5 after intracoronary infusion of acetylcholine and adenosine) (Fig. 1).
Genetic analysis
Protein | Gene | OMIM | SNP ID | N change | Amino acid change | Primers | Ta °C |
---|---|---|---|---|---|---|---|
eNOS |
NOS3
| 163729 | rs1799983 | G-T | Glu298Asp | 5′-CATGAGGCTCAGCCCCAGAAC-3′ | 60 |
5′-AGTCAATCCCTTTGGTGCTCAC-3′ | |||||||
SERCA |
ATP2A2
| 108740 | rs56243033 | G-A | Ala724Ala | 5′-TGAACGATGCTCCTGCTCTG-3′ 5′-TGGGACGAGATGAGGTAGCG-3′ | 60 |
rs12312588 | G-A | Leu734Leu | |||||
Nav1.5 |
SCN5A
| 600163 | rs6599230 | A-G | Ala29Ala | 5′-CCTCTGCTCCATTGACAAGG-3′ 5′-GCAGCCATCGAGAAGCG-3′ | 60 |
rs1805124 | A-G | His558Arg | 5′-TATGAAGCCACGTTCCAGCC-3′ 5′-CCATTGCAGTCCACAGTGC-3′ | ||||
Kir6.1 |
KCNJ8
a
| 600935 | rs11046182 | A-G | Intron 2 | 5′-TCGAGATGAAACTGTTCCACC-3′ 5′-TGCCATCTTATATGGGCATGGC-3′ | 60 |
rs74069151 | A-G | Intron 2 | 5′-AAGAGGTAGGCTGGATAATATCG-3′ 5′-ATTTCCAAATAATGTTTGACCATTC-3′ | ||||
rs77763829 | A-G | Ala171Ala | 5′-AAGTTGGCTAGTCTTTCTGCAAGC-3′ 5′-TTCTTGACCACCTGGATGCGCAC-3′ | ||||
rs16924297 | T-C | Leu265Leu | 5′-AAGCTGTGCTTCATGTTCCG-3′ 5′-TTAACAGTGTTGCCAAATTTGG-3′ | 60 | |||
rs34093632 | G-A | Leu285Leu | |||||
rs35941868 | C-T | Ser312Ser | |||||
rs34130387 | delC | delC315 | |||||
rs34811413 | T-C | Val334Ala | |||||
rs34951653 | delC | delC391 | 5′-TGTCCATTGTGACTGAGGAAGAAGG-3′ 5′-AACAGACTCATTTCTTGACC-3′ | 60 | |||
rs72554071 | C-T | Ser422Leu | |||||
rs7310043 | T-A | 3′UTR | 5′-TTGTTTCATACATGTAGAATTCGC-3′ 5′-TTATGGTATGTCAGGCTTAGATTG-3′ | 60 | |||
rs7309783 | T-A | 3′UTR | |||||
rs7295420 | A-G | 3′UTR | |||||
Kir6.2 |
KCNJ11
a
| 600937 | rs5219 | A-G | Lys23Glu | 5′-GTGGAGGTAAGGAAGAGTCTGG-3′ 5′-AGACGAGAAGGAGTGGATGC-3′ | 59 |
rs140636367 rs5218 | G-A C-T | Ser118Ser Ala190Ala | 5′-AAGTGGCCACACACATTGC-3′ 5′-TCAATGACATGGTAGATGATCAGC-3′ | 59 | |||
rs5216 | C-G | Leu267Leu | 5′-TGGACATCCCCATGGAGAAC-3′ 5′-CTTGTAACACCCTGGATGAGC-3′ | 59 | |||
rs1800467 | C-G | Leu270Val | |||||
rs5215 | G-A | Val337Ile | |||||
rs8175351 | G-A | Lys381Lys | |||||
rs41282930 | C-G | Ser385Cys | |||||
Kv1.5 |
KCN5A
b
| 176267 | rs61737395 | G-T | Gly31Val | 5′-TTCTTGACGTCAGGGCCAAGCG-3′ 5′-CCGGAGATGTTGATGTGGACG-3′ | 60 |
rs71584818 | A-T | Glu33Val | |||||
rs71581015 | 27 bp del | Ser62_Asp72 del | |||||
rs71541953 | C-T | Pro73Ser | |||||
rs71537801 | G-A | Arg87Gln | |||||
ND | C-T | Pro91Leu | |||||
rs17215395 | C-T | Ala115Val | |||||
rs45504599 | C-T | Ser127Ser | 5′-ACCAGGCTCTGGGCACGGCG-3′ 5′-TGGAGATGAGGATAACCAAGACCG-3′ | 60 | |||
rs1056462 | G-C | Gly128Gly | |||||
rs1056463 | T-A | Leu138Gln | |||||
rs41276730 | C-A | Leu185Met | |||||
rs12720444 | C-T | Asn190Asn | |||||
rs12720443 | G-C | Leu205Leu | |||||
rs35853292 | G-C | Glu211Asp | |||||
rs77281462 | C-T | Arg212Cys | |||||
rs3197074 | C-G | Arg214Gly | |||||
rs1056464 | C-T | Pro228Ser | |||||
rs55699243 | T-G | Leu238Leu | |||||
rs55874756 | T-C | Ser248Pro | |||||
rs55958438 | C-A | Arg250Arg | |||||
rs45618444 | C-G | Ala251Gly | |||||
rs17215409 | C-T | Pro307Ser | 5′-TTATCTTCGAGTATCCGGAGAGC-3′ 5′-TGGAGGCCTGCAAGGTCTTG-3′ | 60 | |||
rs17215402 | C-T | Pro310Leu | |||||
rs72546671 | C-T | Leu340Leu | |||||
rs61753194 | T-A | Val341Glu | |||||
rs35130466 | Ins C | InsC367 | |||||
ND | G-T | Glu375Ter | |||||
rs2359641 | T-C | Gly383Gly | |||||
rs76708779 | G-A | Gly384Arg | |||||
rs17221805 | G-A | Leu499Leu | 5′-GTGTTCCGCATCTTCAAGC-3′ 5′-TAGATATCCATGTTCAGCAAGCC-3′ | 60 | |||
ND | C-T | Thr527Met | |||||
rs71582899 | G-A | Pro532Pro | |||||
rs71581016 | G-A | Arg554Gln | |||||
rs71581017 | G-T | Gly568Val | |||||
ND | C-T | Ala576Val | |||||
rs12720445 | G-A | Arg578Lys |
Statistical analysis
Definition of cardiovascular risk factors
Results
Coronary artery disease (n = 155) | Microvascular dysfunction (n = 46) | Normal subjects (n = 41) |
p
| |
---|---|---|---|---|
Age | 66.62 ± 11.935 | 59.38 ± 14.92 | 59.25 ± 10.18 | 0.0001 |
Male gender | 80 % (124/155) | 43.4 % (20/46) | 34.1 % (14/41) | 0.0001 |
Family history of MI | 48.3 % (75/155) | 39.1 % (18/46) | 19.5 % (8/41) | 0.028 |
Hypertension | 78 % (121/155) | 54.3 % (25/46) | 17 % (7/41) | 0.0001 |
Type II diabetes mellitus | 30.3 % (47/155) | 13 % (6/46) | 34.1 % (14/41) | 0.185 |
Dyslipidemia | 51.6 % (80/155) | 32.6 % (15/46) | 41.4 % (17/41) | 0.271 |
Smoking status | 47 % (73/155) | 23.9 % (11/46) | 24.3 % (10/41) | 0.030 |
eNOS/NOS3
| – | – | – | NS |
SERCA/ATP2A2
| – | – | – | NS |
Nav1.5/SCN5A
| – | – | – | NS |
Kir6.1/KCNJ8
| – | – | – | NS |
Kir6.2/KCNJ11: rs5215_GG | 12.2 % (19/155) | 8.6 % (4/46) | 21.9 % (9/41) | 0.041 |
Kir6.2/KCNJ11: rs5219_AA | 10.9 % (17/155) | 10.8 % (5/46) | 21.9 % (9/41) | 0.019 |
Kv1.5/KCNA5
| – | – | – | NS |
Normal subjects (group 3) n = 41 | Coronary artery disease (group 1) n = 155 |
p
| |
---|---|---|---|
(a) | |||
Nav1.5/SCN5A: rs1805124_GG | 19.5 % (8/41) | 7.7 % (12/155) | 0.027 |
Kir6.2/KCNJ11: rs5215_GG | 21.9 % (9/41) | 10.9 % (17/155) | 0.048 |
Normal subjects (group 3) n = 41 | Microvascular dysfunction (group 2) n = 46 |
p
| |
---|---|---|---|
(b) | |||
eNOS/NOS3: rs1799983_GT | 7.3 % (3/41) | 26 % (12/46) | 0.021 |
Kir6.2/KCNJ11: rs5218_CT | 26.8 % (11/41) | 45.6 % (21/46) | 0.048 |
Kir6.2/KCNJ11: rs5219_AA | 21.9 % (9/41) | 8.7 % (4/46) | 0.049 |
p
| OR | 95 % CI | ||
---|---|---|---|---|
(a) | ||||
Age | 0.002 | 1.065 | 1.024 | 1.109 |
Male gender | 0.001 | 6.261 | 2.467 | 15.891 |
Family history of MI | 0.009 | 3.625 | 1.389 | 9.460 |
Hypertension | 0.011 | 3.363 | 1.317 | 8.587 |
Type II diabetes mellitus | 0.865 | 2.599 | 0.255 | 9.585 |
Dyslipidemia | 0.768 | 1.956 | 0.520 | 1.986 |
Smoking status | 0.012 | 3.717 | 1.335 | 10.346 |
Kir6.2/KCNJ11: rs5215_GG | 0.047 | 0.319 | 0.100 | 0.991 |
(b) | ||||
Age | 0.644 | 0.992 | 0.959 | 1.027 |
Male gender | 0.988 | 1.007 | 0.428 | 2.366 |
Family history of MI | 0.209 | 0.550 | 0.217 | 1.396 |
Hypertension | 0.087 | 1.506 | 0.215 | 1.193 |
Type II diabetes mellitus | 0.597 | 1.351 | 0.443 | 4.120 |
Dyslipidemia | 0.065 | 1.227 | 0.525 | 2.896 |
Smoking status | 0.740 | 1.172 | 0.457 | 3.005 |
eNOS/NOS3: rs1799983_GT | 0.023 | 5.234 | 1.260 | 8.521 |
Kir6.2/KCNJ11: rs5219_AA |
0.055
| 0.260 | 0.066 | 1.028 |