MICA polymorphism identified by whole genome array associated with NKG2D-mediated cytotoxicity in T-cell large granular lymphocyte leukemia

Aaron D. Viny, Michael J. Clemente, Monika Jasek, Medhat Askar, Hemant Ishwaran, Amy Nowacki, Aiwen Zhang, Jaroslaw P. Maciejewski

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Background Large granular lymphocyte leukemia is a semi-autonomous clonal proliferation of cytotoxic T cells accompanied by immune cytopenias and various autoimmune conditions. Due to the rarity of this disease and its association with autoimmune diseases, a theoretical germline or somatic mutation might have significant penetrance, thus enabling detection, even from samples of suboptimal size, through genome-wide association studies. Design and Methods To investigate a non-mendelian genetic predisposition to large granular lymphocyte leukemia, we used a step-wise method for gene discovery. First, a modified 'random forests' technique was used for candidate gene identification: this was followed by traditional allele-specific polymerase chain reaction, sequencing modalities, and mechanistic assays. Results Our analysis found an association with MICA, a non-peptide-presenting, tightly regulated, stress-induced MHC-like molecule and cognate receptor for NKG2D, found abundantly on large granular lymphocyte leukemia cells. Sequencing of germline DNA revealed a higher frequency of MICA*00801/A5.1 in patients with large granular lymphocyte leukemia than in matched controls (64% versus 41%, P<0.001, homozygous 40% versus 15%, P<0.001). Flow cytometry was employed to determine the expression of MICA within hematologic compartments, showing that the signal intensity of MICA was increased in granulocytes from neutropenic patients with large granular lymphocyte leukemia in comparison with that in controls (P=0.033). Furthermore, neutrophil counts were inversely correlated with MICA expression (R2=0.50, P=0.035). Finally, large granular lymphocyte leukemia cells were able to selectively kill MICA+ Ba/F3 lymphocytes transfected with human MICA*019 in a dose-dependent manner compared to naïve cells (P<0.001), an effect mitigated by administration of an anti-NKG2D antibody (P=0.033). Conclusions Our results illustrate that MICA-NKG2D played a role in disease pathogenesis in the majority of patients in our cohort of cases of large granular lymphocyte leukemia and further investigation into this signaling axis may provide potent therapeutic targets.

Original languageEnglish
Pages (from-to)1713-1721
Number of pages9
JournalHaematologica
Volume95
Issue number10
DOIs
StatePublished - Oct 1 2010
Externally publishedYes

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Large Granular Lymphocytic Leukemia
Genome
Genetic Association Studies
NK Cell Lectin-Like Receptor Subfamily K
Penetrance
Genome-Wide Association Study
Genetic Predisposition to Disease
DNA Sequence Analysis
Granulocytes
Sample Size
Autoimmune Diseases
Anti-Idiotypic Antibodies
Flow Cytometry
Neutrophils
Alleles
Lymphocytes
T-Lymphocytes
Polymerase Chain Reaction
Mutation

Keywords

  • Genome-wide association study
  • GWAS
  • LGL
  • MICA
  • Neutropenia
  • SNP

ASJC Scopus subject areas

  • Hematology

Cite this

MICA polymorphism identified by whole genome array associated with NKG2D-mediated cytotoxicity in T-cell large granular lymphocyte leukemia. / Viny, Aaron D.; Clemente, Michael J.; Jasek, Monika; Askar, Medhat; Ishwaran, Hemant; Nowacki, Amy; Zhang, Aiwen; Maciejewski, Jaroslaw P.

In: Haematologica, Vol. 95, No. 10, 01.10.2010, p. 1713-1721.

Research output: Contribution to journalArticle

Viny, Aaron D. ; Clemente, Michael J. ; Jasek, Monika ; Askar, Medhat ; Ishwaran, Hemant ; Nowacki, Amy ; Zhang, Aiwen ; Maciejewski, Jaroslaw P. / MICA polymorphism identified by whole genome array associated with NKG2D-mediated cytotoxicity in T-cell large granular lymphocyte leukemia. In: Haematologica. 2010 ; Vol. 95, No. 10. pp. 1713-1721.
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abstract = "Background Large granular lymphocyte leukemia is a semi-autonomous clonal proliferation of cytotoxic T cells accompanied by immune cytopenias and various autoimmune conditions. Due to the rarity of this disease and its association with autoimmune diseases, a theoretical germline or somatic mutation might have significant penetrance, thus enabling detection, even from samples of suboptimal size, through genome-wide association studies. Design and Methods To investigate a non-mendelian genetic predisposition to large granular lymphocyte leukemia, we used a step-wise method for gene discovery. First, a modified 'random forests' technique was used for candidate gene identification: this was followed by traditional allele-specific polymerase chain reaction, sequencing modalities, and mechanistic assays. Results Our analysis found an association with MICA, a non-peptide-presenting, tightly regulated, stress-induced MHC-like molecule and cognate receptor for NKG2D, found abundantly on large granular lymphocyte leukemia cells. Sequencing of germline DNA revealed a higher frequency of MICA*00801/A5.1 in patients with large granular lymphocyte leukemia than in matched controls (64{\%} versus 41{\%}, P<0.001, homozygous 40{\%} versus 15{\%}, P<0.001). Flow cytometry was employed to determine the expression of MICA within hematologic compartments, showing that the signal intensity of MICA was increased in granulocytes from neutropenic patients with large granular lymphocyte leukemia in comparison with that in controls (P=0.033). Furthermore, neutrophil counts were inversely correlated with MICA expression (R2=0.50, P=0.035). Finally, large granular lymphocyte leukemia cells were able to selectively kill MICA+ Ba/F3 lymphocytes transfected with human MICA*019 in a dose-dependent manner compared to na{\"i}ve cells (P<0.001), an effect mitigated by administration of an anti-NKG2D antibody (P=0.033). Conclusions Our results illustrate that MICA-NKG2D played a role in disease pathogenesis in the majority of patients in our cohort of cases of large granular lymphocyte leukemia and further investigation into this signaling axis may provide potent therapeutic targets.",
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T1 - MICA polymorphism identified by whole genome array associated with NKG2D-mediated cytotoxicity in T-cell large granular lymphocyte leukemia

AU - Viny, Aaron D.

AU - Clemente, Michael J.

AU - Jasek, Monika

AU - Askar, Medhat

AU - Ishwaran, Hemant

AU - Nowacki, Amy

AU - Zhang, Aiwen

AU - Maciejewski, Jaroslaw P.

PY - 2010/10/1

Y1 - 2010/10/1

N2 - Background Large granular lymphocyte leukemia is a semi-autonomous clonal proliferation of cytotoxic T cells accompanied by immune cytopenias and various autoimmune conditions. Due to the rarity of this disease and its association with autoimmune diseases, a theoretical germline or somatic mutation might have significant penetrance, thus enabling detection, even from samples of suboptimal size, through genome-wide association studies. Design and Methods To investigate a non-mendelian genetic predisposition to large granular lymphocyte leukemia, we used a step-wise method for gene discovery. First, a modified 'random forests' technique was used for candidate gene identification: this was followed by traditional allele-specific polymerase chain reaction, sequencing modalities, and mechanistic assays. Results Our analysis found an association with MICA, a non-peptide-presenting, tightly regulated, stress-induced MHC-like molecule and cognate receptor for NKG2D, found abundantly on large granular lymphocyte leukemia cells. Sequencing of germline DNA revealed a higher frequency of MICA*00801/A5.1 in patients with large granular lymphocyte leukemia than in matched controls (64% versus 41%, P<0.001, homozygous 40% versus 15%, P<0.001). Flow cytometry was employed to determine the expression of MICA within hematologic compartments, showing that the signal intensity of MICA was increased in granulocytes from neutropenic patients with large granular lymphocyte leukemia in comparison with that in controls (P=0.033). Furthermore, neutrophil counts were inversely correlated with MICA expression (R2=0.50, P=0.035). Finally, large granular lymphocyte leukemia cells were able to selectively kill MICA+ Ba/F3 lymphocytes transfected with human MICA*019 in a dose-dependent manner compared to naïve cells (P<0.001), an effect mitigated by administration of an anti-NKG2D antibody (P=0.033). Conclusions Our results illustrate that MICA-NKG2D played a role in disease pathogenesis in the majority of patients in our cohort of cases of large granular lymphocyte leukemia and further investigation into this signaling axis may provide potent therapeutic targets.

AB - Background Large granular lymphocyte leukemia is a semi-autonomous clonal proliferation of cytotoxic T cells accompanied by immune cytopenias and various autoimmune conditions. Due to the rarity of this disease and its association with autoimmune diseases, a theoretical germline or somatic mutation might have significant penetrance, thus enabling detection, even from samples of suboptimal size, through genome-wide association studies. Design and Methods To investigate a non-mendelian genetic predisposition to large granular lymphocyte leukemia, we used a step-wise method for gene discovery. First, a modified 'random forests' technique was used for candidate gene identification: this was followed by traditional allele-specific polymerase chain reaction, sequencing modalities, and mechanistic assays. Results Our analysis found an association with MICA, a non-peptide-presenting, tightly regulated, stress-induced MHC-like molecule and cognate receptor for NKG2D, found abundantly on large granular lymphocyte leukemia cells. Sequencing of germline DNA revealed a higher frequency of MICA*00801/A5.1 in patients with large granular lymphocyte leukemia than in matched controls (64% versus 41%, P<0.001, homozygous 40% versus 15%, P<0.001). Flow cytometry was employed to determine the expression of MICA within hematologic compartments, showing that the signal intensity of MICA was increased in granulocytes from neutropenic patients with large granular lymphocyte leukemia in comparison with that in controls (P=0.033). Furthermore, neutrophil counts were inversely correlated with MICA expression (R2=0.50, P=0.035). Finally, large granular lymphocyte leukemia cells were able to selectively kill MICA+ Ba/F3 lymphocytes transfected with human MICA*019 in a dose-dependent manner compared to naïve cells (P<0.001), an effect mitigated by administration of an anti-NKG2D antibody (P=0.033). Conclusions Our results illustrate that MICA-NKG2D played a role in disease pathogenesis in the majority of patients in our cohort of cases of large granular lymphocyte leukemia and further investigation into this signaling axis may provide potent therapeutic targets.

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KW - Neutropenia

KW - SNP

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