Studies On The Pathogenesis Of Age-related Macular Degeneration And Its Correlation With Cardiovascular Heart Disease

Objective: Age related macular degeneration(AMD)is a multigenic genetic disease.Age is the primary risk factor for AMD.In this dissertation,single-cell transcriptome sequencing was used to study the changes of retinal pigment epithelium(RPE)and choroid with age,so as to deepen the understanding the effect of age on the pathogenesis of AMD.Through validating e QTL-SNP loci,the population variation related to AMD disease was identified in Sichuan Han population.There is a certain correlation between coronary atherosclerotic heart disease(CHD)and AMD in plaque formation and pathogenesis,the correlation between them have been established through meta-analysis and clinical research in this dissertation.Methods: The retinal pigment epithelium and choroid from donors of seven age points were analyzed by single-cell transcriptome sequencing,and 18 loci were tested by time-of-flight mass spectrometry to find out the genetic loci related to AMD in the Han population in Sichuan,China.The correlation between AMD and CHD and the risk factors associated with them were analyzed by meta-analysis and case-control study.Results: By sequencing the RNA of more than 300000 single cells from human RPE and choroid,10 major RPE and choroidal cell types and their specific expressed genes were identified across macula and its surrounding regions and seven age groups.This study revealed the differences in the types and distribution of RPE and choroidal cells in macula and its surrounding regions.Moreover,the genes related to the expression of NABA core matrix,VEGFA-VEGFR2,and endocytosis were negatively correlated with age,while the genes related to cell division,apoptosis,autophagy and cytokine signal in the immune system were positively correlated with age.NABA core matrix was related to supramolecular fiber tissue pathway.Overexpression of elastin(ELN)could protect elastin from aging related relaxation and enhance the elasticity of fibrous tissue.The intercellular interaction analysis revealed a wide network of connections between RPE and choroidal cell types.The regulators with the greatest magnitude of change related to transcription in old age are IRF7,IRF8,NFKB1,REL and ELF1,which are relevant to immune and inflammatory responses.Through the analysis of single nucleotide variations(SNV),we found that most of the random variations of somatic cells in RPE and choroid were harmless.The association analysis showed that ARMS2-HTRA1 rs11528744(or =1.391,95% CI = 1.179-1.640,P = 0.000),CTRB2-CTRB1 rs9928736(or = 0.695,95%CI = 0.544-0.888,P = 0.004)and B3 GALTL rs4381465(or = 0.614,95% CI = 0.448-0.841,P = 0.002)were associated with AMD,and ARMS2-HTRA1 rs11528744 had the strongest predictive ability for disease(AUC = 0.594,95% CI: 0.561-0.527).Through the analysis of 132039 samples in the observational studies,we found that MI was the risk factor of AMD(RR = 1.12,95% CI = 1.04-1.21,P = 0.005)and AMD was the risk factor of CHD(RR = 1.18,95% CI = 1.15-1.21,P<0.001).In case control analysis,the incidence rate of AMD in CHD and control groups was 5% and 0.90%(P=0.013)respectively,and the incidence rate of CHD in AMD and control groups was 13.4% and 1%(P<0.001)respectively.Multivariate analysis revealed that the risk factors associated with AMD and CHD were age,smoking,gender,hypertension,diabetes and BMI.Conclusion: This dissertation provides a comprehensive single-cell transcriptional map of macula and surrounding regions and human retinal pigment epithelium and choroid at seven ages,and expands and deepens the understanding of the characteristics of human retinal RPE and choroidal cells,and the molecular mechanism of effect of aging on RPE and choroid.This dissertation suggests that ELN may be used as a candidate gene to improve choroidal relaxation and RPE structure,which lays a foundation for the study of the expression of RPE genes and pinpoint to a pontential target for the development of intervention measures for the treatment of aging or eye diseases.The loci associated with the risk of AMD were ARMS2-HTRA1 rs11528744,BCRA1 rs9928736 and B3 GALTL rs4381465.AMD may increase the risk of CHD,and the impact of diabetes on the two risk is more significant than other factors.Gender has an impact on both.