| Objective:In this study,outpatients with short stature were clinically evaluated and data collected by us.According to the criteria for screening,12 patients with suspected single gene mutation were selected for short stature.The whole exome sequencing method was used to screen for possible pathogenic genes and loci.Based on the type and location of the mutated gene,genetic verification was performed through first-generation sequencing,and the harmfulness and pathogenicity of the mutation site were analyzed using bioinformatics related software,thereby revealing the role of ANKRD11 gene mutation in the pathogenesis of patients with short stature.Content:In this study,the whole exon sequencing method was used to conduct genetic studies on suspected single-gene variant short stature patients and their immediate family members in Jining,thereby revealing variant genes related to familial short stature onset.According to the inclusion and exclusion criteria(see the specific research methods for inclusion criteria and exclusion criteria),12 short patients with suspected single gene mutations were screened out,and the patients were included in the electronic medical record system of the affiliated hospital of Jining Medical University to collect clinical data,biochemistry and imaging Data,and collect blood samples of their parents and siblings,and use blood genomic DNA extraction kits to extract DNA,then perform whole exome sequencing on the DNA of the blood samples,verify the mutation sites by first-generation sequencing,and conduct biological information analysis.Method:We conducted a comprehensive medical evaluation based on clinical and genetic characteristics,including the collection of basic information(including ID number,gender,age,birth status,parental height)and physical examination indicators(including height,weight,Tanner stage,abnormal signs).All enrolled patients had perfect blood routine,trigonometry,liver function,kidney function,electrolytes,insulin-like growth factor-1(IGF-1),and insulin-like growth factor binding protein-3(Insulin-like growth factor binding protein-3,IGFBP-3),bone age film,pituitary magnetic resonance imaging,etc.,completed two growth hormone provocation tests including: insulin hypoglycemia growth hormone provocation test and levodopa growth hormone provocation test.According to the inclusion and exclusion criteria,blood samples of 12 patients who were suspected of single gene mutation and their parents and siblings were collected.The DNA was extracted using the blood genomic DNA extraction kit,followed by whole exon sequencing and Sanger sequencing for detection and Identify genetic mutations associated with the patient ’s short stature.Various computer prediction tools and sequence variation libraries further predict the pathogenicity of mutation sites.And one of the patients was followed up for 24 months to observe the growth rate and treatment effect of patients treated with recombinant human growth hormone.Results:According to the function of genes and the genetic pattern of genes leading to diseases,possible pathogenic genes were selected.After the above steps of screening and filtering,a total of 10 genes and 11 mutation sites were selected.According to ACMG’s recommendation,the mutation sites were classified.Among them,only the ANKRD11(c.2579C>T)mutations related to the short stature phenotype and classified as Pathogenic gene mutation sites.A heterozygous point mutation(c.2579C>T)was confirmed in the patient ’s ANKRD11 gene,which was inherited from his mother.The mutation site is located in the highly conserved region of ANKRD11 protein,and it is predicted to be highly pathogenic in computer prediction programs and sequence variation repositories.In addition,patients receiving growth hormone replacement therapy for 24 months showed good height gains.Conclusion:A heterozygous point mutation of AKNRD11 gene was identified in a Chinese patient with short stature phenotype.The patient was treated effectively with growth hormone supplementation. |
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