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Effects of Single Nucleotide Polymorphisms of the β2 Adrenergic Receptor and of Adenylate Cyclase on Sickle Red Cell Adhesion to Laminin.

Publication ,  Conference
Eyler, CE; Jackson, TL; De Castro, LM; Zennadi, R; Vance, JM; Ashley-Koch, A; Telen, MJ
Published in: Blood
November 16, 2004

Single nucleotide polymorphisms (SNPs) of the β2 adrenergic receptor (β2AR) gene (ADRB2) on chromosome 5 have been implicated in clinical variability of several cardiopulmonary disorders. Also, Hoppe et al. [2004] have recently found evidence that polymorphisms of ADRB2 correlate with the incidence of stroke in sickle cell disease (SCD). Stimulation of sickle red cell (SS RBC) adrenergic receptors by epinephrine, as well as stimulation of adenylate cyclase by forskolin, activates the B-CAM/LU laminin receptor [Hines et al. 2003]. There is also evidence that activation of B-CAM/LU on SS RBCs occurs in vivo [Zen et al. 2004]. However, SS RBCs from different individuals show variable baseline adhesion to laminin, as well as variable responsiveness to stimulation by epinephrine. We have found that butoxamine, which specifically inhibits the β2AR, blocked epinephrine-induced SS RBC adhesion by 91%, thus implicating β2ARs in activation of adhesion. Thus, we explored whether SNPs of the genes encoding β2AR (the mediator of epinephrine effects) and adenylate cyclase (a downstream effector of β2AR) affect the degree of baseline adhesion to laminin, as well as the degree to which RBCs upregulate adhesion in response to epinephrine. All samples studied were obtained from non-transfused Hb SS patients in steady state. SS RBCs from 14 of 20 patients studied showed increased adhesion in response to epinephrine, with a mean fold increase of 1.98±0.9. These results are similar to those previously described by Hines. Four SNPs of ADRB2 were examined by direct sequencing; two of these reside in the leader sequence, while the other two encode amino acid substitutions. Four intronic SNPs in ADCY6 were studied by Taqman assays. We found that the arg16gly substitution in β2AR is associated with elevation of baseline SS RBC adhesion measured at 1 dyne/cm2: arg/arg=122, arg/gly=149, gly/gly=262 (mean adherent cells/mm2, n=3, 11, 6 respectively, p=0.03 for gly/gly vs arg/arg + arg/gly). However, the arg16gly substitution was not found to be associated with a statistically significant difference either in the maximal adhesion observed after stimulation by epinephrine: arg/arg=116, arg/gly=263, gly/gly=286 (mean adherent cells/mm2), or in the fold increase observed after stimulation. In addition, two linked SNPs (HCV1244841 and RS3730070, r2=0.8) of ADCY6, which is encoded by chromosome 12, also had a statistically significant effect on baseline but not on epinephrine-stimulated SS RBC adhesion to laminin. For HCV1244841, mean baseline adhesion to laminin at 1 dyne/cm2 was 118 and 303 cells/mm2 for AA+AG and GG, respectively (n=13 and 6, p=0.007), while for RS3730070 mean baseline adhesion was 130 and 327 cells/mm2 for CC+CG and GG, respectively (n=15 and 5, p=0.01). However, these ADCY6 SNPs also showed no statistically significant effect on epinephrine-stimulated adhesion. Overall, both the β2AR and ADCY6 polymorphisms appeared to significantly affect baseline adhesion but not epinephrine-stimulated adhesion in this study. We conclude that β2AR and adenylate cyclase polymorphisms affect red cell adhesive function. We theorize that signaling pathway polymorphisms, by affecting baseline activation of SS RBC adhesion, may influence the severity of SCD.

Duke Scholars

Published In

Blood

DOI

EISSN

1528-0020

ISSN

0006-4971

Publication Date

November 16, 2004

Volume

104

Issue

11

Start / End Page

3565 / 3565

Publisher

American Society of Hematology

Related Subject Headings

  • Immunology
  • 3213 Paediatrics
  • 3201 Cardiovascular medicine and haematology
  • 3101 Biochemistry and cell biology
  • 1114 Paediatrics and Reproductive Medicine
  • 1103 Clinical Sciences
  • 1102 Cardiorespiratory Medicine and Haematology
 

Citation

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MLA
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Eyler, C. E., Jackson, T. L., De Castro, L. M., Zennadi, R., Vance, J. M., Ashley-Koch, A., & Telen, M. J. (2004). Effects of Single Nucleotide Polymorphisms of the β2 Adrenergic Receptor and of Adenylate Cyclase on Sickle Red Cell Adhesion to Laminin. In Blood (Vol. 104, pp. 3565–3565). American Society of Hematology. https://doi.org/10.1182/blood.v104.11.3565.3565
Eyler, Christine E., Terry L. Jackson, Laura M. De Castro, Rahima Zennadi, Jeffery M. Vance, Allison Ashley-Koch, and Marilyn J. Telen. “Effects of Single Nucleotide Polymorphisms of the β2 Adrenergic Receptor and of Adenylate Cyclase on Sickle Red Cell Adhesion to Laminin.” In Blood, 104:3565–3565. American Society of Hematology, 2004. https://doi.org/10.1182/blood.v104.11.3565.3565.
Eyler CE, Jackson TL, De Castro LM, Zennadi R, Vance JM, Ashley-Koch A, et al. Effects of Single Nucleotide Polymorphisms of the β2 Adrenergic Receptor and of Adenylate Cyclase on Sickle Red Cell Adhesion to Laminin. In: Blood. American Society of Hematology; 2004. p. 3565–3565.
Eyler, Christine E., et al. “Effects of Single Nucleotide Polymorphisms of the β2 Adrenergic Receptor and of Adenylate Cyclase on Sickle Red Cell Adhesion to Laminin.Blood, vol. 104, no. 11, American Society of Hematology, 2004, pp. 3565–3565. Crossref, doi:10.1182/blood.v104.11.3565.3565.
Eyler CE, Jackson TL, De Castro LM, Zennadi R, Vance JM, Ashley-Koch A, Telen MJ. Effects of Single Nucleotide Polymorphisms of the β2 Adrenergic Receptor and of Adenylate Cyclase on Sickle Red Cell Adhesion to Laminin. Blood. American Society of Hematology; 2004. p. 3565–3565.

Published In

Blood

DOI

EISSN

1528-0020

ISSN

0006-4971

Publication Date

November 16, 2004

Volume

104

Issue

11

Start / End Page

3565 / 3565

Publisher

American Society of Hematology

Related Subject Headings

  • Immunology
  • 3213 Paediatrics
  • 3201 Cardiovascular medicine and haematology
  • 3101 Biochemistry and cell biology
  • 1114 Paediatrics and Reproductive Medicine
  • 1103 Clinical Sciences
  • 1102 Cardiorespiratory Medicine and Haematology