Identification Of The Causative Mutations For Two Hereditary Nephropathy

Genetic diseases are caused by one or more abnormalities in the genome.They are hereditary,intractable and lifelong.They are often determined at birth(congenital)or acquired.For example,Congenital Stupidity,Polydactyly,Congenital Deafness and Muteness,Hemophilia and so on,these genetic diseases are completely determined by genetic factors,and only after birth for a certain period of time,sometimes it takes several years,more than ten years or even decades to appear obvious symptoms.At present,about 4000 kinds of genetic diseases have been found,and new genetic diseases are found every year.Although genetic diseases are rare,the incidence rate is from several thousand to several million parts,but the harm is extremely serious,which will bring heavy burden to families and society.So far,there is no reliable and effective treatment for genetic diseases,but with the rapid development of cytology and molecular genetics,genetic diagnosis has become the main way of disease prevention and control and targeted therapy.In the first section of the thesis,we explored the significance of VHL in the cellular process,prognosis and drug selection of renal cell carcinoma.The TCGA database downloads information on 539 patients with renal cell carcinoma(RCC),of which 226 have VHL mutations.The survival and drug sensitivity of RCC after VHL mutation were analyzed by cBioPortal and GDSC.The results showed that after VHL mutation,the expression of VHL gene was down-regulated in the tumor tissues of RCC patients,and the drug resistance was changed.The differential expression genes(DEGs)of VHL mutations were further explored,and 22 genes were up-regulated and 670 genes were down-regulated.Using DAVID to analyze the 692 DEGs identified from the functional level,GO enrichment showed that DEGs were significantly enriched mainly in peptidase activity,endopeptidase activity,proteolysis,negative regulation of hydrolase activity,chloride ion transport across membranes,and pH regulation Wait.KEGG enrichment is mainly in retinol metabolism,cytochrome P450 exogenous substance metabolism,chemical carcinogenicity,complement system,etc.Using GESA to explore related functions and pathways,the results showed that the VHL mutation Renin Angiotensin System(RAS)pathway was more active,while the Vibrio Cholerae Infection,Steroid Biosynthesis,and Cardiac Muscle contraction pathways were silent.In order to systematically analyze the related mechanism of VHL mutation,the protein interaction network was screened through the String database,and the hub gene was screened using Cytoscape software.The top 10 genes were identified as central genes,including AHSG,ALB,APOA1,APOA2,SERP INC1,ITIH2,APOA5,SPP2,SERPIND1,KNG1,AHSG have the highest node degree among the central genes.The functional annotation analysis of the three most significant gene modules in the PPI network shows that modules 1-3 are mainly related to cholesterol metabolism,insulin secretion,glucagon signaling pathway,neurotransmitter receptor activity regulation,and signaling Regulating receptor activity.Based on the above results,this study believes that VHL mutations in renal cell carcinoma may affect its prognosis and disease progression.It is more cautious and individualized treatment for RCC patients with VHL mutations.This study has identified the main pathways and genes related to VHL mutations in RCC,and may provide new targets for RCC treatment by formulating treatment strategies for such special subtypes.In the second section of the thesis,a family of 23 people with autosomal dominant polycystic kidney disease was analyzed.The proband’s polycystic kidney disease was diagnosed through the results of clinical examination and B-ultrasonic examination.The clinical data of the family were collected,whole exome sequencing(WES)was used to find the pathogenic mutation in the family,and family members,The genetic variants detected in 100 normal people and 100 end-stage renal disease patients were verified by first-generation sequencing.Bioinformatics analysis was performed on the mutated genes:SIFT and NCBI were used to analyze the pathogenicity and conservatism of the mutated genes to predict the pathogenicity of the mutated sites.The structure of PKD1 and the mutant protein was predicted and mapped using Swiss-Model.This study found that the proband has a new mutation that has not yet been reported.9484delC,further first-generation sequencing results confirmed that 1 patient in this family had the same mutation,while normal members of the family did not have this mutation.Bioinformatics analysis indicated that the frameshift mutation changed the arginine at position 3162 to alanine,and the stop codon was shifted.The protein sequence was shortened from 4302 amino acids to 3314 amino acids,and the protein structure changed greatly,PLAT/LH2 domain is damaged.At the same time,the site area is highly conserved and may cause disease after mutation.This study reported a new mutation site in the PKD1 gene of polycystic kidney disease.Through molecular genetic analysis,the pathogenicity of this site was analyzed to promote the diagnosis,genetic counseling and treatment of polycystic kidney disease.