| (1)Clinical characteristics and gene analysis of fanconi anemia in childrenPurpose:Fanconi anemia is a rare disease with a low incidence.This article retrospectively analyzes the clinical characteristics and gene types of 22 Han children with fanconi anemia in the Children’s Hospital of Soochow University,and analyzes the prognosis of these patients.It hopes to provide clinical data for early diagnosis of such diseases,timing of treatment,and better prognosis.Methods:Collect clinical data of patients with fanconi anemia:general conditions,first symptoms,congenital abnormalities,laboratory tests,diagnosed disease types,severity groups,treatment process and outcome.Use R language for data analysis and use the method chisq.test()to automatically output X-squared chi-squared value and P-value probability.For the case where the theoretical frequency of the chi-square test is too small,the Fisher exact test method is used for data analysis.P<0.05 considered the difference to be statistically significant.Result:A collection of 22 children with fanconi anemia patients in the Children’s Hospital Of Soochow University,all of whom were Han nationality,including 15 mals and 7 females,with a male-to-female ratio of 2.14:1.1 case has not yet involved in abnormal hematopoiesis.The average age of onset of other patients is 4 years and 8 months,and the median age of onset is 4 years and 9 months(4 months to 10 years).The average time between onset and diagnosis was 1 year,and the median time was 2 months(1 month to 8 years).Three patients have family history.There were 14 cases with congenital abnormalities,accounting for 63.6%.There were 10 cases with finger deformities(45%),5 cases with coffee spots(22.7%),2 cases with obvious growth dysplasia,and other cases such as binocular cleft with pulmonary sequestration in 1 case,duodenal diaphragmatic hernia in 1 case,ectopic renal fusion in 1 case,and congenital hypothyroidism in 1 case.The first manifestation was fever,12 cases(54.5%);4 cases with bleeding(18.1%);4 cases with pale complexion(18.1%);4 cases with physical examination(abnormal blood test results).Fever is the most common first sign in children under 5 years old.The genetic testing was completed in 21 patients.A total of 7 FANC genotypes and 42 variant types were detected,and 25 new variant types were found,with complex and diverse gene variants.Among them,13 cases were FANCA genotype,accounting for 61.9%.After analysis,it is found that the deletion of FANCA gene fragments may be related to the younger age of onset(mostly 0-5 years old).The current disease status is manifested in 20 cases of bone marrow failure(BMF)(13 cases of severe and 7 cases of non-severe disease),and 1 case of acute myeloid leukemia(AML)M5a.After analysis,there was no statistically significant difference between the severity of bone marrow failure and the age of onset,congenital malformations,mutant gene types,gene mutation types and transplant or not.Sixteen patients were treated with hematopoietic stem cell transplantation(72.7%),and 6 patients were not treated with transplantation.As of the follow-up day,the average follow-up time was 4 years(0-11 years),2 cases died,3 cases were lost to follow-up,and 17 cases survived.Fourteen patients treated with hematopoietic stem cell transplantation recovered their hematopoietic function.Unfortunately,primary implantation failure occurred in 1 case after hematopoietic stem cell transplantation and another patient died 9 months after transplantation.Among the patients who did not undergo transplantation,1 patient died of severe infection,1 case was not yet ill,3 patients were lost to follow-up,and one was treated with oral androgen.The 6-month overall survival rate(OS)of 22 patients was 94.7%,the 3-year OS was 88.4%,and the 5year OS was 88.4%.Conclusion:FANCA gene mutation was the most common in 22 Han Fanconi anemia children.Patients with FANCA gene fragments deletion have an earlier age of onset of BMF.We have discovered 25 new mutation types,enriching the FA gene mutation spectrum.(Ⅱ)Mechanism of fanconi anemia based on proteomics technology.Objective:Fanconi Anemia(FA)is a kind of congenital aplastic Anemia,which is characterized by congenital bone marrow failure,malformation and malignant tendency,and can develop into acute myeloid leukemia(AML).However,the mechanism of FA develop to AA,even AML is unclear.In this study,we used proteomics technology to explore the mechanisms among them.Methods:In this study,bone marrow from patients with fanconi anemia and aplastic anemia,as well as normal human bone marrow samples and bone marrow samples from patients with fanconi anemia progressing to acute myeloid leukemia were collected.Proteomic data analysis was performed.Differences in bone marrow samples from patients with fanconi anemia and aplastic anemia,fanconi anemia and AML were studied.Using omics data to carry out Gene Ontology analysis,annotation,visualization and comprehensive research.A deep correlation protein-protein interaction(PPI)network was constructed using the STRING website and visualized.Results:Compared with the normal control samples,five differentially expressed proteins were identified in the bone marrow of fanconi anemia patients,including two upregulated proteins and three down-regulated proteins.Compared with the AA sample,a total of 7 differentially expressed proteins,including 2 up-regulated proteins and 5 downregulated proteins,were found in fanconi anemia samples.The up-regulated proteins are mainly concentrated in the regulation of T cell activation,positive regulation of cell adhesion,trans-membrane receptor protein tyrosine kinase signaling pathway,intracellular signal transduction,immune response activation of cell surface receptor signaling pathway,etc.The down-regulated proteins were enriched in the phagocytosis of fC-y receptor signaling pathway,the regulation of stress fiber assembly,and the regulation of actin filament tissue.In addition,contrast bone marrow samples of fanconi anemia with BMF and AML samples of proteomics data,we found 155 differentially expressed proteins(among them,142 upregulated and 13 downregulated proteins)(AML/BMF).The functions affected by upregulated proteins include:leukocyte activation is involved in immune response,bone marrow leukocyte activation,platelet degranulation and actin cytoskeleton reorganization.The functions of down-regulated proteins include coagulation,hemostasis,bicarbonate transport and oxygen binding.Up-regulated protein enrichment signaling pathways includes:pentose phosphate pathway,regulation of actin cytosomes,ribosomes,tight junctions,leukocyte transendothelial migration,and fc-r-mediated phagocytosis.Furthermore,through the constructed protein-protein interaction network,we found that We found that HIST1H1D,HIST1H3A,PSME1 and THRAP3 may play an important role in the progression of fanconi anemia from BMF to AML.In the end of this study,cell experiments confirmed that PSME1 had an important role in the proliferation of leukemia cells.Conclusions:FA showed specific proteomics which involved in abnormal function of T cell immunity,cytoskeleton,and glycometabolism.Overall patient survival analysis showed that high expression of HIST1H1D,Hist1h3a,PSME1 and THRAP3 was associated with poor prognosis of AML in FA patients.(Ⅲ)The analysis of hematopoietic stem cell transplantation on fanconi anemia in childrenPurpose:To explore the timing of hematopoietic stem cell transplantation and how to optimize the conditioning regimen under different donors and different disease states.Reduce acute and chronic graft-versus-host disease(GVHD)and related complications,and provide a reference for improving the prognosis of patients with fanconi anemia.Methods:The clinical data of 16 Han children with fanconi anemia who underwent allogeneic hematopoietic stem cell transplantation(Allo-HSCT)at the Children’s Hospital affiliated Of Soochow University from August 2015 to July 2021 were retrospectively analyzed.The median age of transplantation was 6 years(2 to 15 years).At the time of transplantation,only one patient had acute myeloid leukemia(AML),and the rest showed bone marrow failure(BMF).In the first transplantation,only 1 case of sibling donor transplantation(MSDT)was performed,15 cases underwent alternative donor HSCT,including 7 cases underwent peripheral blood stem cell transplantation(URD-PBSCT)from unrelated donors(4 cases of full compatibility,3 cases of 9/10 HLA-matched);4 cases of unrelated cord blood stem cell transplantation(URD-UCBT)(2 cases with full-matched,2 cases with 8/10 HLA-matched);4 cases with haploid hematopoietic stem cell transplantation(Haplo-HSCT)(1 case with 8/10 HLA-matched 6 cases)/10 HLA matched 2 cases,5/10 HLA matched 1 case),of which 1 case was AML(M5a).Among all patients,2 patients underwent multiple transplants(referring to more than 2 times).Among all patients,2 patients underwent multiple transplantation(more than 2 times),of which case 14 underwent Haplo HSCT for secondary transplantation,15 patients underwent URD-PBSCT for secondary transplantation(the same donor as the first transplantation),but got graft failure again.Then we choose Haplo-HSCT for the third transplantation.Using reduced-dose conditioning regimen:fludarabine(FLU),cyclophosphamide(CTX),rabbit anti-human thymocyte immunoglobulin(ATG),systemic radiotherapy(TBI),busulfan(Bu).Different conditioning regimen were selected according to the total amount of blood transfusion before transplantation,donors and leukemia.The adverse reactions of conditioning regimen,graftversus-host disease,complications,chimerism and treatment outcome were monitored.The follow-up time ranged from 1 month to 6 years,with a median of 4 years.Result:The number of mononuclear cells(MNC)transplanted from sibling donors:22.7 ×108/kg,CD34+ cell number:10.85×106/kg.Median MNC of peripheral blood stem cell transplantation from unrelated donors:8.15(3.87～13.50)× 108/kg,CD34+cell count:7.33(1.77～29.30)× 106/kgMedian MNC of haploid hematopoietic stem cell transplantation:7.47(8.62～10.24)× 108/kg,CD34+cell count:4.31(3.68-14.17)×106/kg;median of MNC for unrelated cord blood stem cell transplantation:0.38(0.14～0.98)×108/kg,CD34+cell count:0.26(0.11～0.84)×106/kg.All 13 cases had sustained engraftment for the first time,3 cases encountered primary graft failure,and 2 cases acquired engraftment after second and third transplantation.The patients acquired engraftment in sibling,haploid and unrelated donor hematopoietic stem cell transplantation with median time of 11 days(7-15 days).The two patients acquired neutropoil engraftment in cord blood stem cell transplantation with time of 15 days and 12 days,while the platelet engraftment with time of time 28 days and 36 days.The incidence of grade Ⅱ-Ⅳ aGVHD was 20%,and one patient developed cGVHD(6.67%).As of the follow-up day,all 15 children were in good condition,and regular monitoring of their condition indicated no progress.The gene mosaic rate of 14 donors and recipients was stable above 95%.One child died of severe infection 9 months after transplantation.The 3year disease-free survival rate was 91.7%.Conclusion:HSCT with low intensity of conditional regimen is still the only way to cure FA hematological abnormalities.Primary engraftment failure of cord blood transplantation is relatively high in FA and alternative donor is the effective option to save engraftment failure.Reducing the complications after transplantation will effectively improve the survival after FA transplantation. |
