Associations of Circulating Extracellular Rnas with Myocardial Remodeling and Heart Failure

Supplementary Online Content

Shah RV, Rong J, Larson MG, et al. Associations of circulating extracellular RNAs with myocardial remodeling and heart failure. Published online August 8, 2018. JAMA Cardiology. doi:10.1001/jamacardio.2018.2371

eMethods. Pathway analysis. eReferences. eTable 1. List of circulating extracellular RNAs included in the study. eTable 2. Age- and sex-adjusted linear models for the association between cardiac structural parameters and extracellular RNAs. eTable 3. Survival analysis for incident HF, Models 1 and 2. eFigure 1. Data overlay on pathway representation of top TGF-beta Signaling Pathway hit. eFigure 2. Data overlay on network representation of top TGF-beta Signaling Pathway hit extended with interactions to hypertension genes.

This supplementary material has been provided by the authors to give readers additional information about their work.

Downloaded From: https://jamanetwork.com/ on 09/28/2021 eMethods. Pathway analysis Gene targets of candidate exRNAs were extracted from experimentally validated interactions in miRTarBase version 6.1 using Target Interaction Finder.1 The output included an xGMML network file that was imported into Cytoscape for subnetwork analysis and data visualization.2 Sub-networks were generated using topological filters to exclude targets of just one exRNA in order to focus on targets with a greater number of candidate exRNA associations and more supporting evidence. Functional characterization of targeted genes was performed using the bioinformatics tool, Enrichr,3 which provided significance scores for enriched processes and pathways from Gene Ontology and WikiPathways4 and disease terms from OMIM in a tabular format.

eReferences 1. Riutta A, Hanspers K, Pico AR. Target interaction finder. GitHub Repository. 2017 2017. 2. Shannon P, Markiel A, Ozier O, et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome research. Nov 2003;13(11):2498-2504. 3. Kuleshov MV, Jones MR, Rouillard AD, et al. Enrichr: a comprehensive gene set enrichment analysis web server 2016 update. Nucleic acids research. Jul 08 2016;44(W1):W90-97. 4. Kutmon M, Riutta A, Nunes N, et al. WikiPathways: capturing the full diversity of pathway knowledge. Nucleic acids research. Jan 04 2016;44(D1):D488-494.

Downloaded From: https://jamanetwork.com/ on 09/28/2021 eTable 1. List of circulating extracellular RNAs included in the study. N specifies number of participants in whom ex-RNA quantification was possible. Mean specifies mean PCR cycle value. SD specifies standard deviation.

Variable N Mean SD piRNA-12151 2751 18.32 1.07 piRNA-54042 2723 16.01 1.61 piRNA-54043 2720 16.05 1.63 miR-16-5p 2716 13.54 1.48 piRNA-2888 2708 16.30 1.57 miR-486-5p 2706 14.83 1.36 miR-126-5p 2698 16.75 1.17 miR-342-3p 2678 17.76 1.10 miR-126-3p 2669 16.07 1.20 miR-30a-5p 2666 16.88 1.20 miR-223-3p 2665 14.72 1.57 miR-451a 2664 11.95 1.89 miR-92a-3p 2664 15.33 1.20 miR-21-5p 2662 15.85 1.40 miR-25-3p 2660 16.94 1.35 miR-19b-3p 2657 15.08 1.29 miR-30e-5p 2654 17.00 1.16 miR-26a-5p 2653 17.28 1.30 miR-191-5p 2652 15.99 1.85 miR-23a-3p 2651 15.88 1.18 miR-221-3p 2648 17.74 1.27 miR-26b-5p 2648 17.40 1.32 miR-30d-5p 2648 17.17 1.37 miR-24-3p 2646 16.50 1.17 miR-15b-5p 2645 16.95 1.16 miR-150-5p 2638 17.22 1.48 miR-19a-3p 2638 15.75 1.32 miR-320a 2634 18.52 1.18 miR-20a-5p 2633 16.83 1.28 miR-484 2633 17.87 1.22 miR-22-3p 2629 16.51 1.33 miR-29a-3p 2627 16.85 1.21 miR-29c-3p 2625 16.86 1.20 snoRNA-1409 2625 18.30 1.44 miR-122-5p 2622 17.90 1.52 miR-93-5p 2612 17.86 1.24 miR-17-5p 2610 17.82 1.23 miR-106b-5p 2609 17.68 1.24 miR-146a-5p 2609 18.42 1.16 snoRNA-1408 2608 18.45 1.42 miR-151b- 2605 18.83 1.13 miR-27a-3p 2603 17.46 1.63 miR-195-5p 2584 16.51 3.14 miR-1260a 2572 18.41 1.30 miR-199a-3p 2561 18.62 1.27 snoRNA-1441 2560 19.36 1.13

Downloaded From: https://jamanetwork.com/ on 09/28/2021 © 2018 American Medical Association. All rights reserved. miR-148a-3p 2556 18.32 1.45 miR-148b-3p 2547 18.55 1.34 miR-222-3p 2531 19.13 1.15 miR-101-3p 2528 18.77 1.23 miR-142-5p 2513 18.68 1.33 piRNA-52468 2510 19.27 1.25 miR-4446-3p 2509 19.27 1.32 snoRNA-1277 2505 18.83 1.34 miR-140-3p 2502 19.19 1.15 miR-28-3p 2471 17.32 1.88 miR-145-5p 2466 19.43 1.18 miR-23b-3p 2458 19.37 1.13 miR-151a-5p 2452 19.37 1.17 miR-185-5p 2448 19.01 1.30 miR-423-5p 2441 19.30 1.20 miR-4433b-5p 2428 19.29 1.21 miR-194-5p 2417 18.51 1.49 miR-186-5p 2412 19.39 1.13 miR-128-3p 2407 19.15 1.34 miR-197-3p 2392 19.93 0.95 miR-130a-3p 2366 19.49 1.17 miR-125a-5p 2338 19.38 1.14 miR-30b-5p 2327 19.65 1.12 miR-125b-5p 2324 19.76 1.06 miR-27b-3p 2312 19.17 1.32 let-7a-5p 2302 19.43 1.24 miR-574-3p 2278 19.54 1.33 let-7b-5p 2277 19.43 1.25 miR-30c-5p 2268 19.75 1.09 miR-103a-3p 2241 19.59 1.16 miR-29b-3p 2237 19.61 1.16 let-7d-5p 2230 19.52 1.23 let-7g-5p 2194 19.44 1.25 miR-99b-5p 2187 19.77 1.24 miR-424-5p 2163 19.16 1.33 miR-6511b-3p 2159 19.93 1.11 miR-18a-5p 2143 19.88 1.07 piRNA-40304 2143 19.08 1.61 let-7i-5p 2136 19.49 1.34 miR-423-3p 2136 19.92 1.11 piRNA-48383 2135 19.60 1.44 let-7d-3p 2129 19.92 1.06 piRNA-43376 2124 19.16 1.60 snoRNA-1460 2087 19.39 1.41 miR-652-3p 2080 19.02 2.00 miR-144-3p 2072 19.01 1.53 piRNA-20101 2051 19.93 1.16 miR-192-5p 2043 19.94 1.11 miR-363-3p 2040 19.53 1.18 miR-28-5p 2025 19.56 1.36 miR-425-5p 2017 19.53 1.22

Downloaded From: https://jamanetwork.com/ on 09/28/2021 © 2018 American Medical Association. All rights reserved. miR-766-3p 2010 19.63 1.05 snoRNA-1401 2010 19.86 1.32 let-7c-5p 1997 20.17 1.01 miR-664b-3p 1997 16.37 3.56 piRNA-57322 1982 19.91 1.12 miR-320b 1978 16.96 3.01 miR-99a-5p 1947 19.53 1.21 miR-532-3p 1935 19.54 1.49 miR-1260b 1912 20.31 0.95 miR-106b-3p 1896 18.08 1.81 miR-494-3p 1880 17.07 2.31 miR-885-5p 1880 19.71 1.34 piRNA-49916 1847 20.09 1.02 miR-15a-5p 1845 20.24 1.00 miR-744-5p 1845 14.11 3.64 let-7f-5p 1830 20.19 1.04 snoRNA-1210 1823 18.57 2.61 miR-139-5p 1793 20.04 1.02 miR-433-3p 1791 20.39 1.12 miR-7977 1779 20.34 1.07 miR-181a-2-3p 1768 19.39 1.50 piRNA-51124 1768 19.78 1.35 piRNA-33872 1736 20.27 1.05 piRNA-58596 1712 20.35 1.03 piRNA-57403 1703 19.84 1.36 miR-301b-3p 1683 18.20 1.96 miR-1246 1676 17.97 2.11 piRNA-33384 1669 20.60 0.83 miR-204-5p 1662 9.72 3.69 piRNA-1340 1634 19.55 1.74 miR-362-3p 1607 20.20 1.13 piRNA-232882 1566 20.27 0.99 piRNA-2962 1561 20.12 1.15 miR-32-5p 1544 20.09 1.20 miR-146b-5p 1531 20.16 1.16 miR-30a-3p 1522 18.03 1.88 piRNA-31112 1507 19.69 1.45 miR-324-5p 1497 20.25 0.85 miR-532-5p 1495 19.50 1.72 miR-432-5p 1478 20.17 1.15 miR-340-5p 1476 20.41 0.99 miR-1271-5p 1463 19.42 1.01 miR-365a-3p 1458 20.42 1.06 snoRNA-1387 1451 20.43 1.13 miR-374b-5p 1450 20.41 1.05 miR-199b-5p 1438 19.89 1.27 piRNA-243353 1426 20.59 1.01 miR-3613-3p 1416 18.41 1.60 piRNA-41647 1408 20.41 0.98 miR-376c-3p 1407 20.38 1.04 miR-4770 1400 20.50 1.05

Downloaded From: https://jamanetwork.com/ on 09/28/2021 © 2018 American Medical Association. All rights reserved. snoRNA-1458 1393 20.16 1.34 miR-34c-5p 1386 20.49 0.97 snoRNA-1472 1385 19.95 1.05 miR-335-5p 1368 20.73 0.85 miR-548e-3p 1362 15.95 2.57 miR-656-3p 1358 20.58 0.95 miR-100-5p 1350 19.11 1.32 piRNA-36772 1343 20.15 1.34 miR-941 1338 18.88 1.16 miR-324-3p 1337 18.31 1.54 snoRNA-1426 1328 20.21 1.29 let-7e-5p 1319 19.13 1.46 snoRNA-1414 1319 20.23 1.30 miR-10b-5p 1317 20.38 1.09 miR-616-5p 1296 17.49 1.25 miR-329-3p 1285 20.49 0.98 piRNA-32519 1276 20.24 1.13 piRNA-57576 1252 20.09 1.30 miR-3615 1236 18.70 1.68 miR-376b-3p 1230 18.25 1.13 piRNA-57581 1230 20.10 1.33 miR-628-3p 1217 18.60 1.03 miR-1180-3p 1211 19.73 0.99 miR-20b-5p 1209 16.87 1.46 miR-564 1207 19.84 1.62 snoRNA-1374 1205 20.42 1.25 miR-92b-3p 1199 16.89 1.68 miR-769-5p 1187 20.58 0.99 miR-151a-3p 1179 18.90 1.30 miR-425-3p 1166 19.40 1.54 miR-542-3p 1161 18.06 1.04 miR-2110 1159 20.45 1.09 miR-29c-5p 1153 20.11 1.28 miR-130b-3p 1142 18.98 1.27 piRNA-265711 1138 21.03 0.97 miR-664a-3p 1133 20.49 1.09 miR-205-5p 1131 20.47 1.12 miR-339-5p 1131 19.92 0.99 miR-660-5p 1124 20.08 1.20 miR-212-3p 1098 20.05 1.16 miR-519b-5p 1081 20.86 0.95 miR-122-3p 1069 19.80 1.11 miR-582-5p 1065 19.27 1.79 miR-4429 1056 20.10 1.04 miR-181b-5p 1055 20.77 0.87 miR-1247-5p 1045 18.07 1.55 miR-181a-3p 1045 19.76 1.44 miR-642a-5p 1044 19.67 1.19 miR-181d-5p 1038 20.69 1.07 miR-154-3p 1034 19.88 1.36 miR-382-3p 1027 19.53 1.81

Downloaded From: https://jamanetwork.com/ on 09/28/2021 © 2018 American Medical Association. All rights reserved. miR-142-3p 1025 19.66 1.18 miR-22-5p 1020 20.41 1.06 miR-377-3p 1018 20.80 0.99 miR-331-3p 1000 20.75 1.05 miR-16-2-3p 997 20.77 0.82 miR-483-5p 992 19.72 1.55 miR-1301-3p 975 20.23 0.92 miR-199a-5p 972 19.55 1.27 miR-143-3p 971 20.01 1.10 miR-582-3p 966 20.44 1.16 miR-375 960 20.45 0.95 snoRNA-1502 957 20.49 1.13 miR-181c-3p 936 20.82 1.05 miR-200b-3p 933 20.79 0.83 miR-381-3p 915 20.15 1.12 miR-144-5p 913 20.88 0.76 let-7b-3p 906 20.50 1.06 miR-1226-3p 904 20.51 1.16 miR-15b-3p 902 20.43 1.11 miR-152-3p 892 20.41 1.10 miR-301a-3p 885 20.65 1.10 miR-33a-3p 881 20.25 1.06 miR-374a-5p 874 20.00 1.13 miR-2355-5p 866 20.47 1.08 miR-361-5p 861 20.67 0.89 miR-323a-3p 853 20.73 1.05 miR-338-3p 849 20.77 0.88 snoRNA-1457 847 20.45 1.30 let-7a-3p 835 20.51 1.08 piRNA-22527 833 19.52 1.99 miR-132-3p 831 20.80 1.05 miR-589-5p 831 20.47 1.07 miR-409-5p 828 20.00 1.34 piRNA-2229 820 20.39 0.94 miR-505-3p 808 20.58 1.12 miR-10a-5p 807 20.86 0.80 miR-483-3p 802 20.75 1.12 miR-486-3p 795 20.64 1.02 miR-181a-5p 792 20.84 0.90 miR-190a-3p 792 19.48 1.44 miR-221-5p 781 17.79 1.68 miR-378a-3p 781 20.37 1.11 piRNA-248758 775 20.09 1.64 miR-1306-5p 765 20.32 1.15 miR-34a-3p 762 20.49 1.38 miR-224-5p 753 20.66 1.11 miR-497-5p 742 20.43 1.15 miR-193b-3p 738 19.73 1.59 miR-223-5p 738 20.80 1.08 miR-590-5p 706 20.29 1.22 snoRNA-1550 697 20.79 1.12

Downloaded From: https://jamanetwork.com/ on 09/28/2021 © 2018 American Medical Association. All rights reserved. miR-193a-5p 690 19.11 2.14 miR-98-5p 685 20.65 1.12 snoRNA-1413 684 20.55 1.34 miR-296-5p 681 20.83 1.12 miR-627-5p 679 20.68 1.15 miR-495-3p 662 20.22 1.22 snoRNA-1384 656 20.91 1.25 miR-214-3p 654 20.84 1.13 miR-181c-5p 634 20.80 0.93 miR-376a-3p 627 20.88 0.83 miR-487b-3p 627 20.58 1.18 piRNA-51374 610 20.71 0.95 miR-320c 606 20.87 1.13 miR-4433a-5p 593 20.51 1.19 miR-345-5p 581 20.74 1.11 miR-133a-3p 568 20.62 1.27 miR-625-3p 564 20.39 1.55 miR-4732-5p 562 20.64 0.94 miR-326 559 20.43 1.22 let-7f-2-3p 551 20.64 1.20 snoRNA-1417 544 20.70 1.25 miR-136-5p 541 20.69 1.22 snoRNA-1563 534 20.49 1.30 snoRNA-1209 528 20.48 1.39 piRNA-36667 526 21.10 0.77 snoRNA-1405 526 20.28 1.53 miR-130b-5p 520 20.74 0.97 miR-215-5p 517 20.81 1.24 miR-485-3p 515 20.79 1.21 snoRNA-1507 513 20.92 1.23 miR-378a-5p 510 20.73 1.19 miR-155-5p 503 20.94 1.11 miR-409-3p 500 20.46 1.26 miR-613 497 20.95 0.94 snoRNA-1394 494 20.82 1.27 miR-584-5p 474 20.69 1.27 piRNA-45809 465 20.47 1.21 miR-1229-3p 447 20.70 1.53 let-7f-1-3p 433 20.87 1.19 miR-342-5p 433 20.73 1.24 miR-550a-3p 430 19.74 1.62 miR-93-3p 429 20.95 1.01 miR-34a-5p 412 21.19 0.62 miR-200c-3p 405 20.96 0.69 miR-337-3p 402 20.65 1.05 miR-206 399 20.85 1.31 snoRNA-1568 395 20.60 1.43 piRNA-227919 380 20.83 1.01 miR-96-5p 377 20.56 1.40 miR-1307-3p 375 20.75 1.15 miR-942-5p 375 20.84 1.30

Downloaded From: https://jamanetwork.com/ on 09/28/2021 © 2018 American Medical Association. All rights reserved. miR-500a-3p 370 20.92 0.96 miR-543 368 20.58 1.08 snoRNA-1291 366 20.54 1.37 miR-17-3p 343 20.71 1.40 piRNA-32636 342 20.70 1.10 piRNA-23216 338 20.94 1.04 miR-1-3p 329 20.49 1.37 miR-4433b-3p 328 20.69 1.18 piRNA-43147 317 20.90 1.03 piRNA-37355 313 20.90 1.09 snoRNA-1407 311 20.19 1.81 miR-652-5p 309 21.05 1.02 miR-659-3p 309 20.73 1.39 piRNA-55662 307 20.65 1.17 piRNA-212993 304 20.24 1.61 piRNA-32637 304 20.59 1.16 miR-182-5p 302 20.93 1.00 piRNA-2096 290 20.93 1.13 piRNA-57387 290 20.82 1.07 miR-24-2-5p 281 20.76 1.50 miR-124-3p 279 20.66 1.38 miR-107 278 20.65 1.45 miR-9-3p 275 20.96 1.03 miR-129-2-3p 272 20.44 1.47 miR-877-5p 271 20.87 1.09 piRNA-40506 271 20.92 1.09 miR-590-3p 270 20.59 1.46 snoRNA-1382 265 20.55 1.62 snoRNA-1403 260 20.43 1.65 piRNA-251099 257 20.76 1.49 miR-145-3p 256 20.84 1.11 miR-503-5p 251 20.56 1.60 miR-545-5p 251 20.54 1.53 snoRNA-1289 243 20.36 1.66 miR-596 241 20.97 1.03 miR-320d 231 20.66 1.13 snoRNA-1290 230 20.68 1.73 piRNA-49867 225 20.28 1.89 piRNA-32212 224 21.05 0.88 miR-7-1-3p 220 20.43 1.61 miR-30e-3p 218 20.55 1.45 miR-7-5p 214 20.78 1.17 piRNA-58593 212 20.95 1.05 piRNA-54782 210 20.78 1.05 miR-654-5p 205 20.88 1.44 snoRNA-1399 205 20.63 1.73 snoRNA-1257 202 20.29 1.73 miR-1304-3p 201 20.71 1.82 miR-576-5p 199 20.48 1.59 miR-654-3p 198 20.90 1.15 piRNA-56396 196 21.08 1.06

Downloaded From: https://jamanetwork.com/ on 09/28/2021 © 2018 American Medical Association. All rights reserved. miR-664a-5p 181 20.25 1.86 miR-21-3p 179 20.45 1.66 miR-98-3p 179 20.33 1.68 miR-1307-5p 178 20.52 1.59 miR-330-3p 178 20.93 1.06 miR-129-5p 177 20.22 1.60 piRNA-41574 175 21.27 1.16 miR-6803-3p 174 20.60 1.35 miR-183-5p 168 20.74 1.26 miR-337-5p 167 20.48 1.80 miR-136-3p 162 20.67 1.22 miR-200a-3p 162 20.71 1.22 piRNA-38142 162 21.30 1.13 snoRNA-1549 162 20.22 1.92 miR-598-3p 161 20.82 1.22 miR-335-3p 160 20.73 1.32 miR-452-5p 159 20.90 1.21 piRNA-40766 157 21.00 1.20 miR-141-3p 156 20.21 1.75 miR-185-3p 152 20.91 1.26 miR-877-3p 151 20.89 1.33 miR-339-3p 150 20.66 1.52 miR-411-3p 147 20.23 1.38 miR-338-5p 145 20.69 1.35 miR-134-5p 140 21.21 1.15 piRNA-36598 138 20.92 1.19 piRNA-218424 136 21.07 1.25 snoRNA-1466 136 20.22 2.07 miR-624-5p 135 20.77 1.31 miR-95-3p 135 20.62 1.48 miR-454-3p 132 20.57 1.32 miR-203a-3p 131 20.11 1.84 miR-424-3p 131 20.34 1.98 piRNA-40039 131 20.51 1.34 snoRNA-1465 126 20.02 2.07 piRNA-46358 123 21.19 0.96 miR-496 121 20.82 1.35 piRNA-44080 113 20.89 2.05 miR-31-3p 111 19.83 1.83 miR-1228-5p 110 20.62 1.74 miR-502-3p 110 20.73 1.26 snoRNA-1490 110 20.01 2.15 piRNA-46251 109 21.18 0.97 miR-501-3p 108 20.64 1.43 snoRNA-1562 108 20.01 2.20 miR-1537-3p 107 20.54 1.44

Downloaded From: https://jamanetwork.com/ on 09/28/2021 © 2018 American Medical Association. All rights reserved. eTable 2. Age- and sex-adjusted linear models for the association between cardiac structural parameters and extracellular RNAs. Each beta coefficient is expressed as the change in log-echo parameter for a two-fold increase in circulating RNA level. All ex-RNAs are significant at a pre- specified FDR rate of 0.05; LVEDV associations had an FDR level of 0.051 and were included here. Ex-RNAs in red are associated with at least 2 phenotypes.

© 2018 American Medical Association. All rights reserved. Left ventricular mass (g/m2.7; log) Left ventricular end-diastolic volume (ml/m2.7; log) Left atrial dimension (log) N Ex-RNA Estimated β P value N Ex-RNA Estimated β P value N Ex-RNA Estimated β P value 2327 miR-30d-5p -0.015 2.68E-06 2314 miR-20a-5p -0.012 2.23E-04 2657 piRNA-54043 -0.007 1.88E-05 2353 miR-342-3p -0.018 2.25E-05 2063 miR-125a-5p -0.014 3.83E-04 2590 miR-26a-5p -0.009 2.31E-05 2334 miR-19b-3p -0.015 2.41E-05 1069 miR-20b-5p -0.015 4.39E-04 2602 miR-223-3p -0.006 2.50E-04 2387 miR-16-5p -0.012 5.46E-05 2297 miR-17-5p -0.012 6.62E-04 2661 piRNA-54042 -0.006 2.60E-04 2328 miR-191-5p -0.010 7.62E-05 2298 miR-93-5p -0.012 7.59E-04 2634 miR-126-5p -0.008 3.46E-04 2369 miR-126-5p -0.015 8.07E-05 2328 miR-26b-5p -0.011 7.90E-04 2501 miR-199a-3p -0.008 4.46E-04 2392 piRNA-54043 -0.011 9.57E-05 2297 miR-106b-5p -0.011 8.97E-04 2586 miR-26b-5p -0.007 5.42E-04 2345 miR-126-3p -0.014 1.90E-04 866 miR-33a-3p -0.015 5.43E-04 2323 miR-24-3p -0.014 2.54E-04 2605 miR-126-3p -0.008 8.49E-04 2311 miR-20a-5p -0.013 3.33E-04 2588 miR-23a-3p -0.008 1.12E-03 2377 miR-486-5p -0.012 3.61E-04 2178 let-7d-5p -0.008 1.15E-03 996 miR-339-5p -0.025 4.52E-04 2331 miR-30e-5p -0.014 4.55E-04 2342 miR-30a-5p -0.013 5.34E-04 2295 miR-93-5p -0.013 6.03E-04 2325 miR-26b-5p -0.012 6.61E-04 1069 miR-20b-5p -0.015 8.93E-04 2322 miR-15b-5p -0.013 9.02E-04 1086 miR-376b-3p -0.020 9.59E-04 2329 miR-26a-5p -0.011 1.19E-03 2294 miR-106b-5p -0.012 1.27E-03 2156 miR-151a-5p -0.013 1.33E-03 2326 miR-221-3p -0.011 1.59E-03 148 piRNA-41574 0.045 1.66E-03 269 piRNA-55662 0.036 1.68E-03 2395 piRNA-54042 -0.009 1.71E-03 2328 miR-23a-3p -0.012 2.30E-03 142 piRNA-40766 0.051 3.27E-03 2294 miR-17-5p -0.011 3.63E-03

Downloaded From: https://jamanetwork.com/ on 09/28/2021 © 2018 American Medical Association. All rights reserved. eTable 3. Multivariable Cox proportional hazards regression models for incident heart failure, as a function of candidate ex-RNAs. Hazard ratios are expressed per a two-fold increase in circulating ex-RNA concentration. Model 1 is a base model adjusted for age, sex, prevalent and time-dependent incident MI; model 2 is a fully adjusted model, including Model 1 covariates plus body mass index, current smoking, systolic blood pressure, use of hypertension medications, and current diabetes. The column labeled “N” indicates the number of subjects included in the Model 1 regression. Abbreviations: HR, hazard ratio; MI, myocardial infarction.

Model 1 Model 2 (age, sex, (Model 1 + Additional risk Ex-RNA N prevalent/incident MI) factors) HR (95% CI) P value HR P value miR-126-5p 2617 0.90 (0.77-1.04) 0.16 - - piRNA-54043 2639 0.99 (0.88-1.11) 0.85 - - miR-126-3p 2591 0.94 (0.81-1.09) 0.39 - - miR-20a-5p 2557 0.84 (0.72-0.98) 0.03 0.86 (0.73-1.00) 0.047 miR-93-5p 2537 0.87 (0.74-1.02) 0.08 - - miR-26b-5p 2570 0.93 (0.80-1.07) 0.30 - - miR-20b-5p 1173 0.92 (0.76-1.10) 0.36 - - miR-26a-5p 2575 0.94 (0.81-1.08) 0.36 - - miR-106b-5p 2534 0.82 (0.71-0.96) 0.01 0.85 (0.73-0.99) 0.04 piRNA-54042 2643 0.99 (0.88-1.11) 0.89 - - miR-23a-3p 2574 0.90 (0.77-1.04) 0.16 - - miR-17-5p 2534 0.82 (0.70-0.96) 0.02 0.84 (0.72-0.99) 0.03

Downloaded From: https://jamanetwork.com/ on 09/28/2021 © 2018 American Medical Association. All rights reserved. eFigure 1. Data overlay on pathway representation of top TGF-beta Signaling Pathway hit. The top pathway hit from the enrichment analysis shown in Figure 2A was the TGF-beta Signaling Pathway from WikiPathways (https://www.wikipathways.org/index.php/Pathway:WP560). The specific miRNA targets enriched in the pathway are indicated in blue. A red border indicates genes in the pathway that interact the miRNA targets enriched in the OMIM term for hypertension (also from Enrichr results) based on GeneMANIA, restricted to default sources of physical, pathway, co-expression and genetic interaction types (i.e., excluding predicted, colocation and others). The pathway and data visualization was performed using Cytoscape.

eFigure 2. Data overlay on network representation of top TGF-beta Signaling Pathway hit extended with interactions to hypertension genes. The same top pathway hit as shown in Supplemental Figure 1, but here represented as a network to explicitly show the additional interactions (green) with hypertension genes (red fill) as implied in both figures by the red border highlights on pathway genes. The source of the content and legend are otherwise identical to Supplemental Figure 1. The network and data visualization was performed using Cytoscape.

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