Positional cloning of quantitative trait nucleotides for blood pressure and cardiac QT-interval by targeted CRISPR/Cas9 editing of a novel long non-coding RNA

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Diseases of the cardiovascular system such as essential hypertension do not have a clear cause, but are known to run in families. The inheritance patterns of essential hypertension and other cardiac diseases suggest that they are not due to a single defective gene but instead are caused by multiple genetic defects that are inherited together in a patient. This complex inheritance makes it difficult to pinpoint the underlying defects. Here, we describe a panel of genetically-engineered rats, using which we have discovered a novel gene, which does not code for any protein, as a gene required for maintenance of normal blood pressure. Structural defects within this non-coding RNA cause hypertension and cardiac short-QT interval. Further, by performing genome surgery to correct the gene defect, we demonstrate the precise error in nucleotides that was inherited and caused hypertension and cardiac short-QT interval syndrome.

Vyšlo v časopise: Positional cloning of quantitative trait nucleotides for blood pressure and cardiac QT-interval by targeted CRISPR/Cas9 editing of a novel long non-coding RNA. PLoS Genet 13(8): e32767. doi:10.1371/journal.pgen.1006961
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1006961

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1. Merai R, Siegel C, Rakotz M, Basch P, Wright J, Wong B, et al. CDC Grand Rounds: A Public Health Approach to Detect and Control Hypertension. MMWR Morb Mortal Wkly Rep. 2016;65(45):1261–1264. doi: 10.15585/mmwr.mm6545a3 27855138

2. Lip GY, Felmeden DC, Li-Saw-Hee FL, Beevers DG. Hypertensive heart disease. A complex syndrome or a hypertensive 'cardiomyopathy'? Eur Heart J. 2000;21(20):1653–1665. doi: 10.1053/euhj.2000.2339 11032692

3. McInnes GT. Hypertension and coronary artery disease: cause and effect. J Hypertens Suppl. 1995;13(2):S49–56. 8576788

4. Clement DL, De Buyzere ML, Duprez DA. Hypertension in peripheral arterial disease. Curr Pharm Des. 2004;10(29):3615–3620. 15579058

5. Kouremenos N, Zacharopoulou IV, Triantafyllidi H, Zacharopoulos GV, Mornos C, Filippatos G, et al. Genes and genetic variations involved in the development of hypertension: focusing on a Greek patient cohort. Hellenic J Cardiol. 2014;55(1):9–16. 24491930

6. Butler MG. Genetics of hypertension. Current status. J Med Liban. 2010;58(3):175–178. 21462849

7. Basson J, Simino J, Rao DC. Between candidate genes and whole genomes: time for alternative approaches in blood pressure genetics. Curr Hypertens Rep. 2012;14(1):46–61. doi: 10.1007/s11906-011-0241-8 22161147

8. Doggrell SA, Brown L. Rat models of hypertension, cardiac hypertrophy and failure. Cardiovasc Res. 1998;39(1):89–105. 9764192

9. Gopalakrishnan K, Kumarasamy S, Yan Y, Liu J, Kalinoski A, Kothandapani A, et al. Increased Expression of Rififylin in A < 330 Kb Congenic Strain is Linked to Impaired Endosomal Recycling in Proximal Tubules. Front Genet. 2012;3:138. doi: 10.3389/fgene.2012.00138 22891072

10. Kumarasamy S, Gopalakrishnan K, Toland EJ, Yerga-Woolwine S, Farms P, Morgan EE, et al. Refined mapping of blood pressure quantitative trait loci using congenic strains developed from two genetically hypertensive rat models. Hypertens Res. 2011;34(12):1263–1270. doi: 10.1038/hr.2011.116 21814219

11. Pillai R, Waghulde H, Nie Y, Gopalakrishnan K, Kumarasamy S, Farms P, et al. Isolation and high-throughput sequencing of two closely linked epistatic hypertension susceptibility loci with a panel of bicongenic strains. Physiol Genomics. 2013;45(16):729–736. doi: 10.1152/physiolgenomics.00077.2013 23757393

12. Nie Y, Kumarasamy S, Waghulde H, Cheng X, Mell B, Czernik PJ, et al. High-resolution mapping of a novel rat blood pressure locus on chromosome 9 to a region containing the Spp2 gene and colocalization of a QTL for bone mass. Physiol Genomics. 2016;48(6):409–419. doi: 10.1152/physiolgenomics.00004.2016 27113531

13. Moreno C, Kaldunski ML, Wang T, Roman RJ, Greene AS, Lazar J, et al. Multiple blood pressure loci on rat chromosome 13 attenuate development of hypertension in the Dahl S hypertensive rat. Physiol Genomics. 2007;31(2):228–235. doi: 10.1152/physiolgenomics.00280.2006 17566075

14. Cowley AW Jr., Moreno C, Jacob HJ, Peterson CB, Stingo FC, Ahn KWet al. Characterization of biological pathways associated with a 1.37 Mbp genomic region protective of hypertension in Dahl S rats. Physiol Genomics. 2014;46(11):398–410. doi: 10.1152/physiolgenomics.00179.2013 24714719

15. Cowley AW Jr., Yang C, Kumar V, Lazar J, Jacob H, Geurts AM, et al. Pappa2 is linked to salt-sensitive hypertension in Dahl S rats. Physiol Genomics. 2016;48(1):62–72. doi: 10.1152/physiolgenomics.00097.2015 26534937

16. Herrera VL, Tsikoudakis A, Ponce LR, Matsubara Y, Ruiz-Opazo N. Sex-specific QTLs and interacting loci underlie salt-sensitive hypertension and target organ complications in Dahl S/jrHS hypertensive rats. Physiol Genomics. 2006;26(3):172–179. doi: 10.1152/physiolgenomics.00285.2005 16720678

17. Herrera VL, Pasion KA, Tan GA, Ruiz-Opazo N. Dahl (S x R) rat congenic strain analysis confirms and defines a chromosome 17 spatial navigation quantitative trait locus to <10 Mbp. PLoS One. 2013;8(2):e58280. doi: 10.1371/journal.pone.0058280 23469157

18. Chauvet C, Charron S, Menard A, Xiao C, Roy J, Deng AY. Submegabase resolution of epistatically interacting quantitative trait loci for blood pressure applicable for essential hypertension. J Hypertens. 2008;26(5):893–901. doi: 10.1097/HJH.0b013e3282f85ded 18398331

19. Deng AY, Chauvet C, Menard A. Alterations in Fibronectin Type III Domain Containing 1 Protein Gene Are Associated with Hypertension. PLoS One. 2016;11(4):e0151399. doi: 10.1371/journal.pone.0151399 27064407

20. Gopalakrishnan K, Morgan EE, Yerga-Woolwine S, Farms P, Kumarasamy S, Kalinoski A, et al. Augmented rififylin is a risk factor linked to aberrant cardiomyocyte function, short-QT interval and hypertension. Hypertension. 2011;57(4):764–771. doi: 10.1161/HYPERTENSIONAHA.110.165803 21357277

21. Saad Y, Yerga-Woolwine S, Saikumar J, Farms P, Manickavasagam E, Toland EJ, et al. Congenic interval mapping of RNO10 reveals a complex cluster of closely-linked genetic determinants of blood pressure. Hypertension. 2007;50(5):891–898. doi: 10.1161/HYPERTENSIONAHA.107.097105 17893371

22. Garrett MR, Dene H, Walder R, Zhang QY, Cicila GT, Assadnia S, et al. Genome scan and congenic strains for blood pressure QTL using Dahl salt-sensitive rats. Genome Res. 1998;8(7):711–723. 9685318

23. Garrett MR, Zhang X, Dukhanina OI, Deng AY, Rapp JP. Two linked blood pressure quantitative trait loci on chromosome 10 defined by dahl rat congenic strains. Hypertension. 2001;38(4):779–785. 11641286

24. Saad Y, Garrett MR, Manickavasagam E, Yerga-Woolwine S, Farms P, Radecki T, et al. Fine-mapping and comprehensive transcript analysis reveals nonsynonymous variants within a novel 1.17 Mb blood pressure QTL region on rat chromosome 10. Genomics. 2007;89(3):343–353. doi: 10.1016/j.ygeno.2006.12.005 17218081

25. Saad Y, Toland EJ, Yerga-Woolwine S, Farms P, Joe B. Congenic mapping of a blood pressure QTL region on rat chromosome 10 using the Dahl salt-sensitive rat with introgressed alleles from the Milan normotensive strain. Mamm Genome. 2008;19(2):85–91. doi: 10.1007/s00335-007-9084-7 18175179

26. Newton-Cheh C, Eijgelsheim M, Rice KM, de Bakker PI, Yin X, Estrada K, et al. Common variants at ten loci influence QT interval duration in the QTGEN Study. Nat Genet. 2009;41(4):399–406. doi: 10.1038/ng.364 19305408

27. Arking DE, Pulit SL, Crotti L, van der Harst P, Munroe PB, Koopmann TT, et al. Genetic association study of QT interval highlights role for calcium signaling pathways in myocardial repolarization. Nat Genet. 2014;46(8):826–836. doi: 10.1038/ng.3014 24952745

28. Gruber AR, Lorenz R, Bernhart SH, Neubock R, Hofacker IL. The Vienna RNA websuite. Nucleic Acids Res. 2008;36(Web Server issue):W70–74. doi: 10.1093/nar/gkn188 18424795

29. Franceschini N, Fox E, Zhang Z, Edwards TL, Nalls MA, Sung YJ, et al. Genome-wide association analysis of blood-pressure traits in African-ancestry individuals reveals common associated genes in African and non-African populations. Am J Hum Genet. 2013;93(3):545–554. doi: 10.1016/j.ajhg.2013.07.010 23972371

30. International Consortium for Blood Pressure Genome-Wide Association S, Ehret GB, Munroe PB, Rice KM, Bochud M, Johnson AD, et al. Genetic variants in novel pathways influence blood pressure and cardiovascular disease risk. Nature. 2011;478(7367):103–109. doi: 10.1038/nature10405 21909115

31. Kato N, Takeuchi F, Tabara Y, Kelly TN, Go MJ, Sim X, et al. Meta-analysis of genome-wide association studies identifies common variants associated with blood pressure variation in east Asians. Nat Genet. 2011;43(6):531–538. doi: 10.1038/ng.834 21572416

32. Levy D, Ehret GB, Rice K, Verwoert GC, Launer LJ, Dehghan A, et al. Genome-wide association study of blood pressure and hypertension. Nat Genet. 2009;41(6):677–687. doi: 10.1038/ng.384 19430479

33. Levy D, Larson MG, Benjamin EJ, Newton-Cheh C, Wang TJ, Hwang SJ, et al. Framingham Heart Study 100K Project: genome-wide associations for blood pressure and arterial stiffness. BMC Med Genet. 2007;8 Suppl 1:S3.

34. Lu X, Wang L, Lin X, Huang J, Charles Gu C, He M, et al. Genome-wide association study in Chinese identifies novel loci for blood pressure and hypertension. Hum Mol Genet. 2015;24(3):865–874. doi: 10.1093/hmg/ddu478 25249183

35. Padmanabhan S, Melander O, Johnson T, Di Blasio AM, Lee WK, Gentilini D, et al. Genome-wide association study of blood pressure extremes identifies variant near UMOD associated with hypertension. PLoS Genet. 2010;6(10):e1001177. doi: 10.1371/journal.pgen.1001177 21082022

36. Salvi E, Kutalik Z, Glorioso N, Benaglio P, Frau F, Kuznetsova T, et al. Genomewide association study using a high-density single nucleotide polymorphism array and case-control design identifies a novel essential hypertension susceptibility locus in the promoter region of endothelial NO synthase. Hypertension. 2012;59(2):248–255. doi: 10.1161/HYPERTENSIONAHA.111.181990 22184326

37. Wain LV, Verwoert GC, O'Reilly PF, Shi G, Johnson T, Johnson AD, et al. Genome-wide association study identifies six new loci influencing pulse pressure and mean arterial pressure. Nat Genet. 2011;43(10):1005–1011. doi: 10.1038/ng.922 21909110

38. Cowley AW Jr. The genetic dissection of essential hypertension. Nat Rev Genet. 2006;7(11):829–840. doi: 10.1038/nrg1967 17033627

39. Rapp JP. Genetic analysis of inherited hypertension in the rat. Physiol Rev. 2000;80(1):135–172. 10617767

40. Gopalakrishnan K, Kumarasamy S, Mell B, Joe B. Genome-wide identification of long noncoding RNAs in rat models of cardiovascular and renal disease. Hypertension. 2015;65(1):200–210. doi: 10.1161/HYPERTENSIONAHA.114.04498 25385761

41. Abiola O, Angel JM, Avner P, Bachmanov AA, Belknap JK, Bennett B, et al. The nature and identification of quantitative trait loci: a community's view. Nat Rev Genet. 2003;4(11):911–916. doi: 10.1038/nrg1206 14634638

42. Winterton SJ, Turner MA, O'Gorman DJ, Flores NA, Sheridan DJ. Hypertrophy causes delayed conduction in human and guinea pig myocardium: accentuation during ischaemic perfusion. Cardiovasc Res. 1994;28(1):47–54. 8068073

43. McIntyre H, Fry CH. Abnormal action potential conduction in isolated human hypertrophied left ventricular myocardium. J Cardiovasc Electrophysiol. 1997;8(8):887–894. 9261715

44. Cheng X, Waghulde H, Mell B, Smedlund K, Vazquez G, Joe B. Pleiotropic Effect of a High Resolution Mapped Blood Pressure QTL on Tumorigenesis. PLoS One. 2016;11(4):e0153519. doi: 10.1371/journal.pone.0153519 27073989

45. Cicila GT, Morgan EE, Lee SJ, Farms P, Yerga-Woolwine S, Toland EJ, et al. Epistatic genetic determinants of blood pressure and mortality in a salt-sensitive hypertension model. Hypertension. 2009;53(4):725–732. doi: 10.1161/HYPERTENSIONAHA.108.126649 19255363

46. Morgan EE, Faulx MD, McElfresh TA, Kung TA, Zawaneh MS, Stanley WC, et al. Validation of echocardiographic methods for assessing left ventricular dysfunction in rats with myocardial infarction. Am J Physiol Heart Circ Physiol. 2004;287(5):H2049–2053. doi: 10.1152/ajpheart.00393.2004 15475530