Differential Expression And Functional Analysis Of LncRNA In MYBPC3 Mutant Familial Hypertrophic Cardiomyopathy

Objective:1 To explore the differential expression of peripheral blood LncRNA and mRNA in FHCM patients subject toMYBPC3 gene mutation;2 To predict the role of biological function and signal transduction pathway of LncRNA with differential expression in HCM through bioinformatic analysis.Method:1 By collecting information about FHCM patients who received treatment in the Department of Cardiology,Fujian Provincial Hospital from Oct.2017 to Oct.2018 as well as their families to obtain basic clinical data and detecting the familial pathogenic genes with the gene-targeted capture high-throughput sequencing technology.3 FHCM families with MYBPC3 gene mutation are concluded by doing so.2 By using Aksomics' Arraystar LncRNA 4.0 expression profile chip to detect the expression level of LncRNA in peripheral blood of the HCM patients and the Health control group so as to determine the differential expression profiles of LncRNA and mRNA in the HCM patients and the control group and screen out the LncRNA with obvious differential expression.3 By conducting GO analysis and KEGG analysis on differentially expressed mRNA and creating a LncRNA-mRNA co-expression network to indicate GO functions of LncRNA and predict the molecular functional mechanism of differentially expressed LncRNA and its possibly engaged biological processes and structural components.4 By conducting correlation analysis?correlation coefficient ?0.9?on obtained LncRNA with obvious differential expression to get co-expressed mRNA and creating a co-expression network to predict its target mRNA and speculate on possibly engaged biological activities or information transmission pathways.Result:1 Three cases of FHCM patents with MYBPC3 gene mutation are collected,of which,two families are affected by c.3624 delC heterozygous deletion mutation in MYBPC3 gene and Family 2 is affected by c.3369₃370insC heterozygous mutation in MYBPC3 gene.In all of the three families,there are members who have been identified as HOCM patients suffering from such complications as syncope and atrial fibrillation indicating poor prognosis,and two of them have a history of sudden death.2 The detection using Arraystar LncRNA 4.0 expression profile chip is made to get the data about differential expression profile of lncRNA and mRNA in the selected patients and the control group.In total,9,961 LncRNAs are featured with up-regulated expression and 14,333 LncRNAs with down-regulated expression,among which,there are 257 up-regulated LncRNAs and 263 down-regulated LcnRNAs where P<0.05 and fold change>1.5.In total,8,100 mRNAs are featured with up-regulated expression and 7,042 mRNAs with down-regulated expression,among which,there are 256 up-regulated mRNAs and 115 down-regulated mRNAs where P<0.05 and fold change>1.5.Adjustment of P?0.05 and LncRNA length<3000 nucleotides,and exclusion of LncRNA not recorded in the relevant database are made to get 12 LncRNAs with significant differential expression?NR₀01566,uc003 mxa.3,ENST00000570919,NR₀28597,NR₀38946,NR₁26010,ENST00000607036,ENST00000428769,ENST00000529298,ENST00000558885,ENST00000412153 and NR₁21660?.3 GO analysis is made on differentially expressed mRNA.For molecular functions,the mRNAs with up-regulated differential expression mainly work to activate phosphatases,N-acetyltransferases,MHC receptors and steroid hormone receptors;for cellular components,they are mainly located in organelles,membrane-bound organelles,cytoplasm,nucleoplasm and catalytic complexes;for biological process,they play a key role in programmed cell death,phospholipid synthesis,protein acetylation and cellular interleukin-4 reaction.On the other hand,the mRNAs with down-regulated differential expression mainly work to activate protein-serine/threonine kinase,long-chain fatty acid-CoA ligase and phosphotransferase,are located in cation channels,chromosomes,nuclear spots and transmembrane transport complexes,and play a main part in folic acid synthesis and long-chain fatty acid-acyl-CoA synthesis.4 The result shows that the differentially expressed up-regulated mRNAs are mainly involved in adherens junction,toxoplasmosis,NF-kappa B signaling pathway,AMPK signaling pathway,terpenoid backbone biosynthesis,cell adhesion molecules,long-term depression,toll-like signal pathway and other signaling pathways,while the down-regulated ones are involved in cGMP-PKG signaling pathway,fatty acid biosynthesis,protein export,fatty acid degradation,fatty acid metabolism and other signaling pathways.5 Correlation analysis is conducted on the selected 12 LncRNAs,resulting in 181 co-expressed mRNAs.With the support of KEGG pathway analysis,it is anticipated that the target mRNAs are involved in glycerophospholipid metabolism,AMPK signaling pathway,adherens junction,long-term depression,glucagon signaling pathway,glycosylphosphatidylinositol?GPI?-anchor biosynthesis and other signaling pathways.LncRNA.Among them,NR₀01566 works in the long-term depression?hsa04730?pathway through the target mRNAs PP2 A and PPP1R17.Conclusion:1 Differential expression of LncRNA and mRNA is found in FHCM patients with MYBPC3 gene mutation and the abovementioned 12 LncRNAs with significant differential expression?NR₀01566,uc003 mxa.3,ENST00000570919,NR₀28597,NR₀38946,NR₁26010,ENST00000607036,ENST00000428769,ENST00000529298,ENST00000558885,ENST00000412153 and NR₁21660?are likely to become diagnostic markers for FHCM and potential targets for gene therapy.2 LncRNA NR₀01566 may play a regulatory role in the process of cardiac hypertrophy through the hsa04730 pathway where its co-expressed mRNAs PPP1R17 and PP2 A are involved.