Methods for treatment with bucindolol based on genetic targeting

Abstract

The present invention concerns the use of methods for evaluating bucindolol treatment for a patient, particularly one with heart failure. It concerns methods for determining whether to administer or prescribe bucindolol to a patient based on whether the patient is homozygous for the Arg 389 polymorphism in the β1-adrenergic receptor (AR).

1 .- 70 . (canceled) 
     
     
         71 . A method for evaluating a difference in response by polymorphic β 1  adrenergic receptor (AR) polypeptides comprising:
 obtaining cells transfected with a nucleic acid encoding a β 1 AR having a glycine at position 389 (β 1 AR Gly389) and incubating β 1 AR Gly389-expressing cells with bucindolol; 
 measuring cAMP accumulation in β 1 AR Gly389-expressing cells after incubation with bucindolol; 
 obtaining cells transfected with a nucleic acid encoding a β 1 AR having an arginine at position 389 (β 1 AR Arg389) and contacting β 1 AR Arg389-expressing cells with bucindolol; 
 measuring cAMP accumulation in β 1 AR Arg389-expressing cells after incubation with bucindolol; and, 
 comparing the levels of cAMP accumulation in 131AR Gly389-expressing cells and β 1 AR Arg389-expressing cells. 
 
     
     
         72 . The method of  claim 72 , wherein some of the β 1 AR Gly389-expressing cells and β 1 AR Arg389-expressing cells that are incubated with bucindolol are also incubated with norepinephrine, and the method further comprises:
 measuring cAMP accumulation in β 1 AR Gly389-expressing cells after incubation with bucindolol and norepineprhine; 
 measuring cAMP accumulation in β 1 AR Arg389-expressing cells after incubation with bucindolol and norepinephrine; and, 
 comparing the levels of cAMP accumulation in β 1 AR Gly389-expressing cells and β 1 AR Arg389-expressing cells exposed to both bucindolol and norepinephrine. 
 
     
     
         73 . The method of  claim 72 , wherein the amount of norepinephrine cells are incubated with is 10 10 μM. 
     
     
         74 . The method of  claim 71 , further comprising determining expression levels of β 1 AR in the β 1 AR Gly389-expressing cells and β 1 AR Arg389-expressing cells. 
     
     
         75 . The method of  claim 74 , wherein expression is determined using radioligand binding. 
     
     
         76 . The method of  claim 71 , wherein the levels of cAMP accumulation are measured using [ 3 H]cAMP and column chromatography. 
     
     
         77 . A method for evaluating differences in propanol effects on expression of proteins in ventricles of the heart from different β 1 AR polymorphisms comprising
 administering propanol to a transgenic mouse expressing human β 1 AR Arg389 and a transgenic mouse expressing human β 1 AR Gly389; 
 measuring expression in the ventricles of both the β 1 AR Arg389-expressing mouse and the β 1 AR-Gly389-expressing mouse of one or more of the following proteins: Gαs, Gαi2, G-protein coupled receptor kinase-2 (GRK2), adenylyl cyclase type 5 (AC5), total phospholamban (T-PLN), phosphorylated phospholamban (P-PLN) and sarcoplasmic endoplasmic reticulum calcium ATPase-2A (SERCA). 
 
     
     
         78 . The method of  claim 77 , wherein propanol is administered to the transgenic mice in drinking water with a concentration of 0.5 mg/ml. 
     
     
         79 . The method of  claim 77 , wherein propanol is administered to the transgenic mice for 6 months. 
     
     
         80 . The method of  claim 77 , wherein protein expression is measured using Western blotting.