ANALYSIS OF CLONAL IMMUNOGLOBULIN AND T-CELL RECEPTORGENE REARRANGEMENTS IN ACUTE MYELOID LEUKEMIA
(Abstract release date: 05/19/16)
EHA Library. Huang Y. 06/09/16; 134533; PB1633
Dr. Ying Huang
Contributions
Contributions
Abstract
Abstract: PB1633
Type: Publication Only
Background
Acute myeloid leukemia (AML), the most common form of leukemia, carries a high mortality rate and economic burden. In 2015, approximately 39,130 individuals were diagnosed in the United States and Europe with roughly half that number dying from the disease (1,2). With the advent of more sensitive molecular assays, the complex architecture and functional heterogeneity of AML has become appreciated though not yet fully elucidated. Fundamentally, understanding the hematopoietic stem cell (HSC) self-renewal and differentiation model will aid in this goal. Initial hierarchical hematopoietic models focused on the lymphoid and myeloid lineage groupings to be segregated. Recently studies suggest that lineage commitment of hematopoietic progenitors may be both multidirectional and reversible with changes in lineage caused by both intrinsic and environmental factors (3). Though AML is classified as a myeloid neoplasm, we were interested in determining the prevalence of clonal rearrangements within the immunoglobulin heavy (IGH) and light chain (IGK), T-cell receptor gamma (TRG) loci in AML patient samples.
Aims
Assess the frequency of IGH, IGK, and TRG rearrangements in AML patient samples.
Methods
DNA was extracted by QIAcube from a random sampling of 200 AML anonymized patient residual peripheral blood (PB) or bone marrow (BM) specimens. DNA was quantified by NanoDrop and normalized. Each DNA sample was tested for 6 different PCR master mixes: IdentiClone IGH Tubes A, B, C, which target the framework (FR) 1, 2, and 3 regions, respectively; IGK Tube A, IGK Tube B, and TRG 2.0. Amplicon products were analyzed using the ABI 3500 XL instrument. Based on the florescent signals, clonal (positive) or polyclonal (negative) were assessed per instructions of use accompanying the IdentiClone assay.
Results
The IdentiClone IGH assay identified 23 (12%), 14 (7%) and 16 (8%) clonal positive samples for FR 1, 2 and 3, respectively. Combining all three IGH tubes increased the clonal detection rate to 28 (14%) with 172 (86%) samples determined to be negative. The IdentiClone IGK assays identified 39 (20%) and 29 (15%) clonal positive samples for Tube A and Tube B, respectively. Combining the two IGK tubes increased clonal detection to 56 (28%), with 143 (72%) determined to be negative. Combining all 5 IGH + IGK tubes, 65 (33%) clonal positive samples and 135 (68%) clonal negative samples were identified. The TRG 2.0 assay detected 85 (43%) clonal positive samples and 114 (57%) clonal negative samples. Overall, using 6 tubes of PCR MM across IGH, IGK and TRG assays, 113 (57%) samples were identified as clonal positive and 87 (44%) samples were identified as clonal negative.
Conclusion
Over 50% of AML samples demonstrated at least one clonal IG or TCR gene rearrangement. Although clonal rearrangements occur in individuals over 60 years of age, clonal rearrangement assays combined with outcome data for AML patients may provide clinical utility and further elucidate functional heterogeneity.
Session topic: E-poster
Keyword(s): AML, Clonality, Ig and TCR gene rearrangement
Type: Publication Only
Background
Acute myeloid leukemia (AML), the most common form of leukemia, carries a high mortality rate and economic burden. In 2015, approximately 39,130 individuals were diagnosed in the United States and Europe with roughly half that number dying from the disease (1,2). With the advent of more sensitive molecular assays, the complex architecture and functional heterogeneity of AML has become appreciated though not yet fully elucidated. Fundamentally, understanding the hematopoietic stem cell (HSC) self-renewal and differentiation model will aid in this goal. Initial hierarchical hematopoietic models focused on the lymphoid and myeloid lineage groupings to be segregated. Recently studies suggest that lineage commitment of hematopoietic progenitors may be both multidirectional and reversible with changes in lineage caused by both intrinsic and environmental factors (3). Though AML is classified as a myeloid neoplasm, we were interested in determining the prevalence of clonal rearrangements within the immunoglobulin heavy (IGH) and light chain (IGK), T-cell receptor gamma (TRG) loci in AML patient samples.
Aims
Assess the frequency of IGH, IGK, and TRG rearrangements in AML patient samples.
Methods
DNA was extracted by QIAcube from a random sampling of 200 AML anonymized patient residual peripheral blood (PB) or bone marrow (BM) specimens. DNA was quantified by NanoDrop and normalized. Each DNA sample was tested for 6 different PCR master mixes: IdentiClone IGH Tubes A, B, C, which target the framework (FR) 1, 2, and 3 regions, respectively; IGK Tube A, IGK Tube B, and TRG 2.0. Amplicon products were analyzed using the ABI 3500 XL instrument. Based on the florescent signals, clonal (positive) or polyclonal (negative) were assessed per instructions of use accompanying the IdentiClone assay.
Results
The IdentiClone IGH assay identified 23 (12%), 14 (7%) and 16 (8%) clonal positive samples for FR 1, 2 and 3, respectively. Combining all three IGH tubes increased the clonal detection rate to 28 (14%) with 172 (86%) samples determined to be negative. The IdentiClone IGK assays identified 39 (20%) and 29 (15%) clonal positive samples for Tube A and Tube B, respectively. Combining the two IGK tubes increased clonal detection to 56 (28%), with 143 (72%) determined to be negative. Combining all 5 IGH + IGK tubes, 65 (33%) clonal positive samples and 135 (68%) clonal negative samples were identified. The TRG 2.0 assay detected 85 (43%) clonal positive samples and 114 (57%) clonal negative samples. Overall, using 6 tubes of PCR MM across IGH, IGK and TRG assays, 113 (57%) samples were identified as clonal positive and 87 (44%) samples were identified as clonal negative.
Conclusion
Over 50% of AML samples demonstrated at least one clonal IG or TCR gene rearrangement. Although clonal rearrangements occur in individuals over 60 years of age, clonal rearrangement assays combined with outcome data for AML patients may provide clinical utility and further elucidate functional heterogeneity.
Session topic: E-poster
Keyword(s): AML, Clonality, Ig and TCR gene rearrangement
Abstract: PB1633
Type: Publication Only
Background
Acute myeloid leukemia (AML), the most common form of leukemia, carries a high mortality rate and economic burden. In 2015, approximately 39,130 individuals were diagnosed in the United States and Europe with roughly half that number dying from the disease (1,2). With the advent of more sensitive molecular assays, the complex architecture and functional heterogeneity of AML has become appreciated though not yet fully elucidated. Fundamentally, understanding the hematopoietic stem cell (HSC) self-renewal and differentiation model will aid in this goal. Initial hierarchical hematopoietic models focused on the lymphoid and myeloid lineage groupings to be segregated. Recently studies suggest that lineage commitment of hematopoietic progenitors may be both multidirectional and reversible with changes in lineage caused by both intrinsic and environmental factors (3). Though AML is classified as a myeloid neoplasm, we were interested in determining the prevalence of clonal rearrangements within the immunoglobulin heavy (IGH) and light chain (IGK), T-cell receptor gamma (TRG) loci in AML patient samples.
Aims
Assess the frequency of IGH, IGK, and TRG rearrangements in AML patient samples.
Methods
DNA was extracted by QIAcube from a random sampling of 200 AML anonymized patient residual peripheral blood (PB) or bone marrow (BM) specimens. DNA was quantified by NanoDrop and normalized. Each DNA sample was tested for 6 different PCR master mixes: IdentiClone IGH Tubes A, B, C, which target the framework (FR) 1, 2, and 3 regions, respectively; IGK Tube A, IGK Tube B, and TRG 2.0. Amplicon products were analyzed using the ABI 3500 XL instrument. Based on the florescent signals, clonal (positive) or polyclonal (negative) were assessed per instructions of use accompanying the IdentiClone assay.
Results
The IdentiClone IGH assay identified 23 (12%), 14 (7%) and 16 (8%) clonal positive samples for FR 1, 2 and 3, respectively. Combining all three IGH tubes increased the clonal detection rate to 28 (14%) with 172 (86%) samples determined to be negative. The IdentiClone IGK assays identified 39 (20%) and 29 (15%) clonal positive samples for Tube A and Tube B, respectively. Combining the two IGK tubes increased clonal detection to 56 (28%), with 143 (72%) determined to be negative. Combining all 5 IGH + IGK tubes, 65 (33%) clonal positive samples and 135 (68%) clonal negative samples were identified. The TRG 2.0 assay detected 85 (43%) clonal positive samples and 114 (57%) clonal negative samples. Overall, using 6 tubes of PCR MM across IGH, IGK and TRG assays, 113 (57%) samples were identified as clonal positive and 87 (44%) samples were identified as clonal negative.
Conclusion
Over 50% of AML samples demonstrated at least one clonal IG or TCR gene rearrangement. Although clonal rearrangements occur in individuals over 60 years of age, clonal rearrangement assays combined with outcome data for AML patients may provide clinical utility and further elucidate functional heterogeneity.
Session topic: E-poster
Keyword(s): AML, Clonality, Ig and TCR gene rearrangement
Type: Publication Only
Background
Acute myeloid leukemia (AML), the most common form of leukemia, carries a high mortality rate and economic burden. In 2015, approximately 39,130 individuals were diagnosed in the United States and Europe with roughly half that number dying from the disease (1,2). With the advent of more sensitive molecular assays, the complex architecture and functional heterogeneity of AML has become appreciated though not yet fully elucidated. Fundamentally, understanding the hematopoietic stem cell (HSC) self-renewal and differentiation model will aid in this goal. Initial hierarchical hematopoietic models focused on the lymphoid and myeloid lineage groupings to be segregated. Recently studies suggest that lineage commitment of hematopoietic progenitors may be both multidirectional and reversible with changes in lineage caused by both intrinsic and environmental factors (3). Though AML is classified as a myeloid neoplasm, we were interested in determining the prevalence of clonal rearrangements within the immunoglobulin heavy (IGH) and light chain (IGK), T-cell receptor gamma (TRG) loci in AML patient samples.
Aims
Assess the frequency of IGH, IGK, and TRG rearrangements in AML patient samples.
Methods
DNA was extracted by QIAcube from a random sampling of 200 AML anonymized patient residual peripheral blood (PB) or bone marrow (BM) specimens. DNA was quantified by NanoDrop and normalized. Each DNA sample was tested for 6 different PCR master mixes: IdentiClone IGH Tubes A, B, C, which target the framework (FR) 1, 2, and 3 regions, respectively; IGK Tube A, IGK Tube B, and TRG 2.0. Amplicon products were analyzed using the ABI 3500 XL instrument. Based on the florescent signals, clonal (positive) or polyclonal (negative) were assessed per instructions of use accompanying the IdentiClone assay.
Results
The IdentiClone IGH assay identified 23 (12%), 14 (7%) and 16 (8%) clonal positive samples for FR 1, 2 and 3, respectively. Combining all three IGH tubes increased the clonal detection rate to 28 (14%) with 172 (86%) samples determined to be negative. The IdentiClone IGK assays identified 39 (20%) and 29 (15%) clonal positive samples for Tube A and Tube B, respectively. Combining the two IGK tubes increased clonal detection to 56 (28%), with 143 (72%) determined to be negative. Combining all 5 IGH + IGK tubes, 65 (33%) clonal positive samples and 135 (68%) clonal negative samples were identified. The TRG 2.0 assay detected 85 (43%) clonal positive samples and 114 (57%) clonal negative samples. Overall, using 6 tubes of PCR MM across IGH, IGK and TRG assays, 113 (57%) samples were identified as clonal positive and 87 (44%) samples were identified as clonal negative.
Conclusion
Over 50% of AML samples demonstrated at least one clonal IG or TCR gene rearrangement. Although clonal rearrangements occur in individuals over 60 years of age, clonal rearrangement assays combined with outcome data for AML patients may provide clinical utility and further elucidate functional heterogeneity.
Session topic: E-poster
Keyword(s): AML, Clonality, Ig and TCR gene rearrangement
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