NEXT GENERATION TARGETED SEQUENCING IMPROVED THE PRECISE DIAGNOSIS OF PEDIATRIC APLASTIC ANEMIA /INHERITED BONE MARROW FAILURE SYNDROMES
(Abstract release date: 05/19/16)
EHA Library. An W. 06/11/16; 135231; S475
Dr. Wenbin An
Contributions
Contributions
Abstract
Abstract: S475
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 12:15 - 12:30
Location: Room H5
Background
Inherited bone marrow failure syndromes（IBMF）is a group of diseases with heterogeneous clinical and genetic characteristics and overlapping symptoms. Diagnosis of IBMFS depend on classic clinical manifestation like early onset, physical anomalies associated, family history of cancer and/or bone marrow history, and abnormal laboratory test including chromosome breakage testing (MMC and/or DEB), mutation analyses and bone marrow chromosome analyses. At present, more than 70 pathogenic gene mutations had been identified. However, in some patients, physical anomalies is absent or delayed, and were diagnosed as acquired bone marrow failure. Therefore, precise genetic analysis is very important for establish a specific diagnosis, predict cancer risk, direct treatment and genetic counseling.
Aims
In this study, we focus on the application of next generation targeted sequencing in precise diagnosis of pediatric aplastic anemia (AA)/IBMF, and compared the results with clinical manifestation and chromosome breakage testing.
Methods
We designed a targeted capture next generation sequencing (NGS) assay to test a panel of 417 blood disease genes. Pediatic patients (≤14 year old) with suspected diagnosis of AA/IBMF were enrolled. Peripheral blood (PB) was used to genetic analysis and oral epithelia cells or PB from the parents were used to identify whether the gene mutations were somatic mutations or not. The results were validated by Sanger sequencing.
Results
We validated the assay using 30 samples with known mutations. The mean depth for targeted sequencing was 200×, 93.28% of targeted regions were covered with >10×.288 patients with pediatric IBMF were enrolled， including 177 subjects were clinically diagnosed as acquired AA, 49 Fanconi anemia (FA), 26 DBA, 8 DC, 15 SCN, 4 congenital thrombocytopenia, 1 Shwachman-Diamond syndrome (SDS). Totally, 66 (23.6%) subjects have IBMF related genetic mutations. In subjects clinically diagnosed as acquired AA, 17 (9.6%) subjects have IBMF related gene mutations, including 10 subjects finally genetically diagnosed as FA, 3 DC, 2 Epstein syndromes, 1 Wiskott-Aldrich syndrome (WAS). In subjects clinically diagnosed as FA, 21 (42.1%) have FANC related gene mutations and 3 (6.9%) have other IBMF related gene mutations. In subjects clinically diagnosed as DC, 7 (85.7%) have telomere related gene mutations. In subjects clinically diagnosed as DBA, 11(37.9%) have ribosomal protein related gene mutations and 1 FA. In subjects clinically diagnosed as SCN, 5 (33.3%) have SCN-related gene mutations, 1 DC, 1 FA and 1 WHIM syndrome, 1 AKT, 1 GFI1 mutation. In subjects clinically diagnosed as congenital thrombocytopenia, 1 CAMT.In subjects finally genetically diagnosed as AA, FA or DC, 4.4%, 18.2% and 9.1% have family history of BMF or malignant blood diseases respectively, and 3.8%, 9.1%, 0% have family history of solid tumor respectively. In subjects finally genetically diagnosed as AA, FA or DC, 8.8%, 66.75%, 63.3% have physical anomalies respectively. Compared to subjects with AA, subjects with FA or DC are more likely to have physical anomalies of short stature and development retardation, Cafe au lait spots of the skin and finger(toe) malformation (24.2% vs. 0%，P=0.000；21.2% vs.1.2%，P=0.021；36.4% vs.1.2%，P=0.000). Moreover, 8.1% subjects present physical anomalies before or at the same time with bone marrow failure. In subjects finally diagnosed as FA or DC, only 27% and 14% have positive MMC respectively.
Conclusion
Compared to traditional diagnostic methods, the targeted NGS analysis is economic, convenient, easy to update and very important for differential diagnosis, personal treatment and follow-up and genetic counselling， especially for make differential diagnosis between acquired and congenital BMF. Moreover, the targeted NGS can be used to make precise diagnosis between IBMFs which have overlapping clinical symptoms. In subjects with IBMF, the positive rate of MMC is low. Therefore, further gene mutation analysis is necessary for highly suspicious subjects with IBMF who have negative result of MMC.
Session topic: Bone marrow failure syndromes incl. PNH - Biology
Keyword(s): Bone marrow failure, Diagnosis, Gene array
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 12:15 - 12:30
Location: Room H5
Background
Inherited bone marrow failure syndromes（IBMF）is a group of diseases with heterogeneous clinical and genetic characteristics and overlapping symptoms. Diagnosis of IBMFS depend on classic clinical manifestation like early onset, physical anomalies associated, family history of cancer and/or bone marrow history, and abnormal laboratory test including chromosome breakage testing (MMC and/or DEB), mutation analyses and bone marrow chromosome analyses. At present, more than 70 pathogenic gene mutations had been identified. However, in some patients, physical anomalies is absent or delayed, and were diagnosed as acquired bone marrow failure. Therefore, precise genetic analysis is very important for establish a specific diagnosis, predict cancer risk, direct treatment and genetic counseling.
Aims
In this study, we focus on the application of next generation targeted sequencing in precise diagnosis of pediatric aplastic anemia (AA)/IBMF, and compared the results with clinical manifestation and chromosome breakage testing.
Methods
We designed a targeted capture next generation sequencing (NGS) assay to test a panel of 417 blood disease genes. Pediatic patients (≤14 year old) with suspected diagnosis of AA/IBMF were enrolled. Peripheral blood (PB) was used to genetic analysis and oral epithelia cells or PB from the parents were used to identify whether the gene mutations were somatic mutations or not. The results were validated by Sanger sequencing.
Results
We validated the assay using 30 samples with known mutations. The mean depth for targeted sequencing was 200×, 93.28% of targeted regions were covered with >10×.288 patients with pediatric IBMF were enrolled， including 177 subjects were clinically diagnosed as acquired AA, 49 Fanconi anemia (FA), 26 DBA, 8 DC, 15 SCN, 4 congenital thrombocytopenia, 1 Shwachman-Diamond syndrome (SDS). Totally, 66 (23.6%) subjects have IBMF related genetic mutations. In subjects clinically diagnosed as acquired AA, 17 (9.6%) subjects have IBMF related gene mutations, including 10 subjects finally genetically diagnosed as FA, 3 DC, 2 Epstein syndromes, 1 Wiskott-Aldrich syndrome (WAS). In subjects clinically diagnosed as FA, 21 (42.1%) have FANC related gene mutations and 3 (6.9%) have other IBMF related gene mutations. In subjects clinically diagnosed as DC, 7 (85.7%) have telomere related gene mutations. In subjects clinically diagnosed as DBA, 11(37.9%) have ribosomal protein related gene mutations and 1 FA. In subjects clinically diagnosed as SCN, 5 (33.3%) have SCN-related gene mutations, 1 DC, 1 FA and 1 WHIM syndrome, 1 AKT, 1 GFI1 mutation. In subjects clinically diagnosed as congenital thrombocytopenia, 1 CAMT.In subjects finally genetically diagnosed as AA, FA or DC, 4.4%, 18.2% and 9.1% have family history of BMF or malignant blood diseases respectively, and 3.8%, 9.1%, 0% have family history of solid tumor respectively. In subjects finally genetically diagnosed as AA, FA or DC, 8.8%, 66.75%, 63.3% have physical anomalies respectively. Compared to subjects with AA, subjects with FA or DC are more likely to have physical anomalies of short stature and development retardation, Cafe au lait spots of the skin and finger(toe) malformation (24.2% vs. 0%，P=0.000；21.2% vs.1.2%，P=0.021；36.4% vs.1.2%，P=0.000). Moreover, 8.1% subjects present physical anomalies before or at the same time with bone marrow failure. In subjects finally diagnosed as FA or DC, only 27% and 14% have positive MMC respectively.
Conclusion
Compared to traditional diagnostic methods, the targeted NGS analysis is economic, convenient, easy to update and very important for differential diagnosis, personal treatment and follow-up and genetic counselling， especially for make differential diagnosis between acquired and congenital BMF. Moreover, the targeted NGS can be used to make precise diagnosis between IBMFs which have overlapping clinical symptoms. In subjects with IBMF, the positive rate of MMC is low. Therefore, further gene mutation analysis is necessary for highly suspicious subjects with IBMF who have negative result of MMC.
Session topic: Bone marrow failure syndromes incl. PNH - Biology
Keyword(s): Bone marrow failure, Diagnosis, Gene array
Abstract: S475
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 12:15 - 12:30
Location: Room H5
Background
Inherited bone marrow failure syndromes（IBMF）is a group of diseases with heterogeneous clinical and genetic characteristics and overlapping symptoms. Diagnosis of IBMFS depend on classic clinical manifestation like early onset, physical anomalies associated, family history of cancer and/or bone marrow history, and abnormal laboratory test including chromosome breakage testing (MMC and/or DEB), mutation analyses and bone marrow chromosome analyses. At present, more than 70 pathogenic gene mutations had been identified. However, in some patients, physical anomalies is absent or delayed, and were diagnosed as acquired bone marrow failure. Therefore, precise genetic analysis is very important for establish a specific diagnosis, predict cancer risk, direct treatment and genetic counseling.
Aims
In this study, we focus on the application of next generation targeted sequencing in precise diagnosis of pediatric aplastic anemia (AA)/IBMF, and compared the results with clinical manifestation and chromosome breakage testing.
Methods
We designed a targeted capture next generation sequencing (NGS) assay to test a panel of 417 blood disease genes. Pediatic patients (≤14 year old) with suspected diagnosis of AA/IBMF were enrolled. Peripheral blood (PB) was used to genetic analysis and oral epithelia cells or PB from the parents were used to identify whether the gene mutations were somatic mutations or not. The results were validated by Sanger sequencing.
Results
We validated the assay using 30 samples with known mutations. The mean depth for targeted sequencing was 200×, 93.28% of targeted regions were covered with >10×.288 patients with pediatric IBMF were enrolled， including 177 subjects were clinically diagnosed as acquired AA, 49 Fanconi anemia (FA), 26 DBA, 8 DC, 15 SCN, 4 congenital thrombocytopenia, 1 Shwachman-Diamond syndrome (SDS). Totally, 66 (23.6%) subjects have IBMF related genetic mutations. In subjects clinically diagnosed as acquired AA, 17 (9.6%) subjects have IBMF related gene mutations, including 10 subjects finally genetically diagnosed as FA, 3 DC, 2 Epstein syndromes, 1 Wiskott-Aldrich syndrome (WAS). In subjects clinically diagnosed as FA, 21 (42.1%) have FANC related gene mutations and 3 (6.9%) have other IBMF related gene mutations. In subjects clinically diagnosed as DC, 7 (85.7%) have telomere related gene mutations. In subjects clinically diagnosed as DBA, 11(37.9%) have ribosomal protein related gene mutations and 1 FA. In subjects clinically diagnosed as SCN, 5 (33.3%) have SCN-related gene mutations, 1 DC, 1 FA and 1 WHIM syndrome, 1 AKT, 1 GFI1 mutation. In subjects clinically diagnosed as congenital thrombocytopenia, 1 CAMT.In subjects finally genetically diagnosed as AA, FA or DC, 4.4%, 18.2% and 9.1% have family history of BMF or malignant blood diseases respectively, and 3.8%, 9.1%, 0% have family history of solid tumor respectively. In subjects finally genetically diagnosed as AA, FA or DC, 8.8%, 66.75%, 63.3% have physical anomalies respectively. Compared to subjects with AA, subjects with FA or DC are more likely to have physical anomalies of short stature and development retardation, Cafe au lait spots of the skin and finger(toe) malformation (24.2% vs. 0%，P=0.000；21.2% vs.1.2%，P=0.021；36.4% vs.1.2%，P=0.000). Moreover, 8.1% subjects present physical anomalies before or at the same time with bone marrow failure. In subjects finally diagnosed as FA or DC, only 27% and 14% have positive MMC respectively.
Conclusion
Compared to traditional diagnostic methods, the targeted NGS analysis is economic, convenient, easy to update and very important for differential diagnosis, personal treatment and follow-up and genetic counselling， especially for make differential diagnosis between acquired and congenital BMF. Moreover, the targeted NGS can be used to make precise diagnosis between IBMFs which have overlapping clinical symptoms. In subjects with IBMF, the positive rate of MMC is low. Therefore, further gene mutation analysis is necessary for highly suspicious subjects with IBMF who have negative result of MMC.
Session topic: Bone marrow failure syndromes incl. PNH - Biology
Keyword(s): Bone marrow failure, Diagnosis, Gene array
Type: Oral Presentation
Presentation during EHA21: On Saturday, June 11, 2016 from 12:15 - 12:30
Location: Room H5
Background
Inherited bone marrow failure syndromes（IBMF）is a group of diseases with heterogeneous clinical and genetic characteristics and overlapping symptoms. Diagnosis of IBMFS depend on classic clinical manifestation like early onset, physical anomalies associated, family history of cancer and/or bone marrow history, and abnormal laboratory test including chromosome breakage testing (MMC and/or DEB), mutation analyses and bone marrow chromosome analyses. At present, more than 70 pathogenic gene mutations had been identified. However, in some patients, physical anomalies is absent or delayed, and were diagnosed as acquired bone marrow failure. Therefore, precise genetic analysis is very important for establish a specific diagnosis, predict cancer risk, direct treatment and genetic counseling.
Aims
In this study, we focus on the application of next generation targeted sequencing in precise diagnosis of pediatric aplastic anemia (AA)/IBMF, and compared the results with clinical manifestation and chromosome breakage testing.
Methods
We designed a targeted capture next generation sequencing (NGS) assay to test a panel of 417 blood disease genes. Pediatic patients (≤14 year old) with suspected diagnosis of AA/IBMF were enrolled. Peripheral blood (PB) was used to genetic analysis and oral epithelia cells or PB from the parents were used to identify whether the gene mutations were somatic mutations or not. The results were validated by Sanger sequencing.
Results
We validated the assay using 30 samples with known mutations. The mean depth for targeted sequencing was 200×, 93.28% of targeted regions were covered with >10×.288 patients with pediatric IBMF were enrolled， including 177 subjects were clinically diagnosed as acquired AA, 49 Fanconi anemia (FA), 26 DBA, 8 DC, 15 SCN, 4 congenital thrombocytopenia, 1 Shwachman-Diamond syndrome (SDS). Totally, 66 (23.6%) subjects have IBMF related genetic mutations. In subjects clinically diagnosed as acquired AA, 17 (9.6%) subjects have IBMF related gene mutations, including 10 subjects finally genetically diagnosed as FA, 3 DC, 2 Epstein syndromes, 1 Wiskott-Aldrich syndrome (WAS). In subjects clinically diagnosed as FA, 21 (42.1%) have FANC related gene mutations and 3 (6.9%) have other IBMF related gene mutations. In subjects clinically diagnosed as DC, 7 (85.7%) have telomere related gene mutations. In subjects clinically diagnosed as DBA, 11(37.9%) have ribosomal protein related gene mutations and 1 FA. In subjects clinically diagnosed as SCN, 5 (33.3%) have SCN-related gene mutations, 1 DC, 1 FA and 1 WHIM syndrome, 1 AKT, 1 GFI1 mutation. In subjects clinically diagnosed as congenital thrombocytopenia, 1 CAMT.In subjects finally genetically diagnosed as AA, FA or DC, 4.4%, 18.2% and 9.1% have family history of BMF or malignant blood diseases respectively, and 3.8%, 9.1%, 0% have family history of solid tumor respectively. In subjects finally genetically diagnosed as AA, FA or DC, 8.8%, 66.75%, 63.3% have physical anomalies respectively. Compared to subjects with AA, subjects with FA or DC are more likely to have physical anomalies of short stature and development retardation, Cafe au lait spots of the skin and finger(toe) malformation (24.2% vs. 0%，P=0.000；21.2% vs.1.2%，P=0.021；36.4% vs.1.2%，P=0.000). Moreover, 8.1% subjects present physical anomalies before or at the same time with bone marrow failure. In subjects finally diagnosed as FA or DC, only 27% and 14% have positive MMC respectively.
Conclusion
Compared to traditional diagnostic methods, the targeted NGS analysis is economic, convenient, easy to update and very important for differential diagnosis, personal treatment and follow-up and genetic counselling， especially for make differential diagnosis between acquired and congenital BMF. Moreover, the targeted NGS can be used to make precise diagnosis between IBMFs which have overlapping clinical symptoms. In subjects with IBMF, the positive rate of MMC is low. Therefore, further gene mutation analysis is necessary for highly suspicious subjects with IBMF who have negative result of MMC.
Session topic: Bone marrow failure syndromes incl. PNH - Biology
Keyword(s): Bone marrow failure, Diagnosis, Gene array
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