Construction And Application Of The Patient Registry Study Database Of Dust Mite Allergic Asthma In Children

PART1 CONSTRUCTION THE PATIENT REGISTRY STUDY DATABASE OF DUST MITE ALLERGIC ASTHMA IN CHILDRENBackground: Asthma is a heterogeneous disease with different phenotypes,endotypes and response to treatment.Dust mite allergic asthma is the most common type of asthma in children.Compared with randomized controlled trial(RCT),patient registry study(PRS)can reflect the real physical condition,clinical diagnosis and treatment process of children more comprehensively.Objective: By constructing the patient registry study database of dust mite allergic asthma in children,we can do clustering analysis for children with different clinical phenotypes.At the same time,we can use the blood samples in the database to further study the different pathophysiology mechanism,so as to understand the causes of children’s different reactions to treatment.Then we can provide more individualized and targeted treatment for children.Methods: According to the method of developing patient registry databases,we constructed the database of dust mite allergic asthma in children.From August 2018 to February 2021,children visited the asthma clinic of the Children’s Hospital of Chongqing Medical University,who met the inclusion criteria and agreed to participate,were enrolled in.Clinical information,laboratory tests and peripheral blood samples were collected after the informed consent was signed.Serum,plasma and peripheral blood mononuclear cells(PBMC)were isolated from peripheral blood.Serum IL-4,IL-5,IL-13,IFN-γ and other common inflammatory factors were detected.The rest of the blood,serum,plasma and PBMC were frozen at-80℃ in the laboratory’s refrigerator or the biological sample bank of the hospital.Results: So far,259 children have been enrolled in.Preliminary analysis of existing data found that there were 164 male and 95 female children.54.83% of the children had a family history of allergic diseases,48.26% had cigarette exposure,and 69.88% were delivered by cesarean section.The median of total IgE(t IgE)was 469.00 k U/L,ratio of Dermatophagoides pteronyssinus(Der P)specific IgE to total IgE(Der p s IgE/t IgE)and Dermatophagoides farina(Der F)specific IgE to total IgE(Der f s IgE/t IgE)were 11.77% and 14.00%,respectively.The median of eosinophil percentage(E%)was 7%.all above the normal value.According to the c-ACT score after ICS treatment,the children were divided into asthma controlled group and uncontrolled group.Preliminary comparison of the indicators between the two groups,it was found that the levels of t IgE,E%,Fe NO and Hb in the uncontrolled group were higher than in the controlled group,while the levels of FEV1% and IL-17 in the uncontrolled group were lower.Conclusions: We have preliminarily constructed a registered study database of children with dust mite allergic asthma.The database will continue to be expanded and improved in the future.By clustering analysis different children and using the clinical blood samples in the database,we can further study the pathophysiological mechanism of different children and the causes of different responses to treatment,so as to provide more individualized and targeted treatment for children.PART2 APPLICATION THE PATIENT REGISTRY STUDY DATABASE OF DUST MITE ALLERGIC ASTHMA IN CHILDRENSECTION 1 THE ROLE OF FLNA IN ALLERGIC ASTHMAAND ITS RESPONSE TO ICSBackground: Exhaled nitric oxide(Fe NO)can accurately reflect airway eosinophilic inflammation,and can be used to predict the response to inhaled corticosteroids(ICS).According to the Fe NO level after ICS treatment,the children enrolled in our datebase were divided into two groups.The differentially expressed genes on peripheral blood eosinophils of the two groups were obtained by RNA-seq.The bioinformatics analysis of RNA-seq results showed that the sharpest gene change between the two groups was filamin A(FLNA),which expression in persistent airway inflammation group was lower than in remission group.At present,there is no research on the role of FLNA in the pathogenesis of asthma or response to ICS treatment.Objective: To explore the role of FLNA in allergic asthma and its response to ICS.Methods: The results of RNA-seq were analyzed by Cytoscape.The gene FLNA with the sharpest change in expression was selected.Verified the results of RNA-seq on PBMC of asthmatic children with airway inflammation remission and persistent after ICS treatment.Analyse the correlation between FLNA and other clinical indicators,laboratory indicators and differentially expressed genes.At the same time,the asthmatic mouse model was sensitized by HDM.The expression of FLNA in the lung tissue was compared with that in the control mice.Results: The expression of FLNA in PBMC of children with allergic asthma after ICS treatment was consistent with that of RNA-seq.The expression of FLNA in persistent airway inflammation group was lower than in remission group(p = 0.0449).In the correlation analysis with other clinical indicators,laboratory indicators and differentially expressed genes,it was positively correlated with FEV1%(r = 0.499,p = 0.03);Negatively correlated with the expression of ITGB4(r = 0.417,p = 0.043)and with the expression of ITGB7(r = 0.488,p = 0.015).In the lung of mice,the expression of FLNA in the asthma group was lower than that in the control group(p = 0.0004),and the expression of ITGB7 in the asthma group was higher than that in the control group(p = 0.0105).FLNA was negatively correlated with ITGB7(r = 0.845,p = 0.0082),but positively correlated with ITGB4(r = 0.608,p = 0.110).Conclusion: FLNA may be involved in airway inflammation and response to ICS in children with allergic asthma by acting on ITGB7.SECTION 2 ANALYSIS OF EXON SEQUENCING IN CHILDREN WITH ALLERGIC ASTHMAAND ABNORMAL ELEVATED SERUM IGEBackground: High IgE syndrome(HIES)is a rare primary immunodeficiency disease.The serum IgE level of patients with this disease is significantly increased,usually more than 2000 Ku/L.These patients may only show asthma,eczema and other atopic symptoms before diagnosis.Diagnosis needs gene screening,but the cost of gene screening is still relatively expensive at present.In our database,we found a small number of children whose total serum IgE was more than 2000 Ku/L.Whether these asthmatic children need to consider HIES and perfect the genetic test,there is no relevant research on this problem now.Objective: To determine whether the children with allergic asthma,who has the abnormally elevated serum IgE,requires gene screening for high IgE syndrome routinely.Methods: The children in our datebase with serum t IgE > 2000 Ku/L were screened out from August 2018 to December 2019.The data of the children were analyzed,and the blood was sent to do whole exon sequencing.Screen out the mutation gene associated with high IgE syndrome from OMIM database and literatures,and find out whether there is mutation in exon sequencing results.The results of exon sequencing were verified by one generation sequencing in patients with mutation gene.Results: From August 2018 to December 2019,there were 7 children with serum t IgE > 2000 Ku/L in our database.Among them,4 cases had heterozygous mutation gene of high IgE syndrome,and 3 cases had no mutation gene of high IgE syndrome.Further,the first generation sequencing did not detect the mutations suggested by exon sequencing in 4children.Conclusion: For children with allergic asthma whose serum IgE is abnormally elevated,routine gene screening is not necessary when there are no other clinical manifestations related to high IgE syndrome.