Clinical Trial Results. org Based on the Iron (Fe) and Atherosclerosis Study (FeAST) Leo R. Zacharski, MD; Bruce K. Chow, MS; Paula S. Howes, MS, APRN; Galina Shamayeva, MS; John A. Baron, MD; Ronald L. Dalman, MD; David J. Malenka, MD; C. Keith Ozaki, MD; and Philip W. Lavori, PhD Published in The Journal of the American Medical Association February 14, 2007 Reduction of Iron Stores and Cardiovascular Outcomes in Patients with Peripheral Arterial Disease A Randomized Controlled Trial

Clinical Trial Results. org FeAST Trial: Background Accumulation of iron in excess of physiologic requirements has been implicated in risk of cardiovascular disease because of increased iron- catalyzed free radical-mediated oxidative stress.Accumulation of iron in excess of physiologic requirements has been implicated in risk of cardiovascular disease because of increased iron- catalyzed free radical-mediated oxidative stress. The objective of this study was to test the hypothesis that reducing body iron stores through phlebotomy will influence outcomes in a cohort of patients with symptomatic peripheral arterial disease (PAD).The objective of this study was to test the hypothesis that reducing body iron stores through phlebotomy will influence outcomes in a cohort of patients with symptomatic peripheral arterial disease (PAD). Accumulation of iron in excess of physiologic requirements has been implicated in risk of cardiovascular disease because of increased iron- catalyzed free radical-mediated oxidative stress.Accumulation of iron in excess of physiologic requirements has been implicated in risk of cardiovascular disease because of increased iron- catalyzed free radical-mediated oxidative stress. The objective of this study was to test the hypothesis that reducing body iron stores through phlebotomy will influence outcomes in a cohort of patients with symptomatic peripheral arterial disease (PAD).The objective of this study was to test the hypothesis that reducing body iron stores through phlebotomy will influence outcomes in a cohort of patients with symptomatic peripheral arterial disease (PAD). Zacharski et al. JAMA; 2007 Feb. 297(6):

Clinical Trial Results. org FeAST Trial: Study Design  Primary Endpoint: All-cause mortality  Secondary Endpoint: Death plus nonfatal myocardial infarction and stroke  Primary Endpoint: All-cause mortality  Secondary Endpoint: Death plus nonfatal myocardial infarction and stroke Control Group n=641 Control Group n=641 Iron-Reduction Group* n=636 Iron-Reduction Group* n= patients with symptomatic but stable PAD Multicenter. Randomized. Controlled. Single-Blinded. Analysis by intent-to-treat. Patients were stratified by hospital, age, and baseline smoking status, diagnosis of diabetes mellitus, ratio of high-density to low-density lipoprotein cholesterol level, and ferritin level. Excluding those with conditions likely to cause acute-phase increase of the ferritin level or with a diagnosis of visceral malignancy within the preceding 5 years 1277 patients with symptomatic but stable PAD Multicenter. Randomized. Controlled. Single-Blinded. Analysis by intent-to-treat. Patients were stratified by hospital, age, and baseline smoking status, diagnosis of diabetes mellitus, ratio of high-density to low-density lipoprotein cholesterol level, and ferritin level. Excluding those with conditions likely to cause acute-phase increase of the ferritin level or with a diagnosis of visceral malignancy within the preceding 5 years R Zacharski et al. JAMA; 2007 Feb. 297(6): * Phlebotomy reduced iron stores with removal of defined volumes of blood at 6 mo. intervals Up to 3.5 years follow-up

Clinical Trial Results. org Outcome Total (n=1277) Control(n=641) Iron Reduction (n=636) HR (95% CI) P value Primary Endpoint 273 (21.4) 148 (23.1) 125 (19.7) 0.85 ( ) 0.17 Secondary Endpoint 385 (30.1) 205 (32) 180 (28.3) 0.88 ( ) 0.20 MI 119 (9.3) 58 (9) 61 (9.6) 1.01 ( ) 0.95 Stroke 61 (4.8) 29 (4.5) 32 (5) 1.22 ( ) 0.46 FeAST Trial: Primary and Secondary Outcome Events Zacharski et al. JAMA; 2007 Feb. 297(6): Abbreviations: CI, confidence interval; HR, hazard ratio; MI, myocardial infarction Comparison of Control and Iron-Reduction Groups for Primary (All-Cause Mortality) and Secondary (Death Plus Nonfatal MI and Stroke) Outcome Events

Clinical Trial Results. org FeAST Trial: Kaplan-Meir Analysis of Primary Endpoint (All-Cause Mortality) No statistically significant differences between treatment groups was observed for the primary endpoint of all- cause mortality (HR 0.85; 95% CI ; p=0.17).No statistically significant differences between treatment groups was observed for the primary endpoint of all- cause mortality (HR 0.85; 95% CI ; p=0.17). Zacharski et al. JAMA; 2007 Feb. 297(6):

Clinical Trial Results. org FeAST Trial: Kaplan-Meir Analysis of Secondary Endpoint (Death Plus Nonfatal MI, or Stroke) There were no statistically significant differences between treatment groups for the secondary endpoint (HR 0.88; 95% CI ; p=0.20).There were no statistically significant differences between treatment groups for the secondary endpoint (HR 0.88; 95% CI ; p=0.20). Neither the cumulative incidence nor the time to occurrence of other nonfatal peripheral, coronary, and cerebral vascular events during the follow-up differed between groups.Neither the cumulative incidence nor the time to occurrence of other nonfatal peripheral, coronary, and cerebral vascular events during the follow-up differed between groups. Zacharski et al. JAMA; 2007 Feb. 297(6):

Clinical Trial Results. org FeAST Trial: Limitations Because of lower than expected accrual, the study was underpowered overall and particularly underpowered to definitively assess outcomes in younger patients and smokers.Because of lower than expected accrual, the study was underpowered overall and particularly underpowered to definitively assess outcomes in younger patients and smokers. This study was single-blinded, and primary and secondary endpoints were adjudicated by a committee external to the study; nonetheless, concerns remain about possible bias, particularly in subgroup analyses.This study was single-blinded, and primary and secondary endpoints were adjudicated by a committee external to the study; nonetheless, concerns remain about possible bias, particularly in subgroup analyses. Because of lower than expected accrual, the study was underpowered overall and particularly underpowered to definitively assess outcomes in younger patients and smokers.Because of lower than expected accrual, the study was underpowered overall and particularly underpowered to definitively assess outcomes in younger patients and smokers. This study was single-blinded, and primary and secondary endpoints were adjudicated by a committee external to the study; nonetheless, concerns remain about possible bias, particularly in subgroup analyses.This study was single-blinded, and primary and secondary endpoints were adjudicated by a committee external to the study; nonetheless, concerns remain about possible bias, particularly in subgroup analyses. Zacharski et al. JAMA; 2007 Feb. 297(6):

Clinical Trial Results. org FeAST Trial: Limitations (cont.) Patients with very high ferritin levels were excluded from the study, and the efficacy of iron reduction in individuals with extreme levels of iron stores is unknown.Patients with very high ferritin levels were excluded from the study, and the efficacy of iron reduction in individuals with extreme levels of iron stores is unknown. Zacharski et al. JAMA; 2007 Feb. 297(6):

Clinical Trial Results. org FeAST Trial: Summary Preplanned analyses of the primary (all-cause mortality) and the secondary (death plus nonfatal MI and stroke) endpoints performed on the entire study cohort showed no effect of iron reduction.Preplanned analyses of the primary (all-cause mortality) and the secondary (death plus nonfatal MI and stroke) endpoints performed on the entire study cohort showed no effect of iron reduction. However, there was a significant interaction with age (1of 5 prespecified biological stratifying factors), suggesting that a beneficial effect might exist in younger patients, and observation that coincides with findings of others.However, there was a significant interaction with age (1of 5 prespecified biological stratifying factors), suggesting that a beneficial effect might exist in younger patients, and observation that coincides with findings of others. Preplanned analyses of the primary (all-cause mortality) and the secondary (death plus nonfatal MI and stroke) endpoints performed on the entire study cohort showed no effect of iron reduction.Preplanned analyses of the primary (all-cause mortality) and the secondary (death plus nonfatal MI and stroke) endpoints performed on the entire study cohort showed no effect of iron reduction. However, there was a significant interaction with age (1of 5 prespecified biological stratifying factors), suggesting that a beneficial effect might exist in younger patients, and observation that coincides with findings of others.However, there was a significant interaction with age (1of 5 prespecified biological stratifying factors), suggesting that a beneficial effect might exist in younger patients, and observation that coincides with findings of others. Zacharski et al. JAMA; 2007 Feb. 297(6):

Clinical Trial Results. org FeAST Trial: Summary (cont.) The FeAST data show that it should be possible to test definitively whether controlling iron levels may reduce disease risk and additional research is needed to further define ferrotoxic diseases, stratifying risk reduction with intervention, and clarify mechanisms, especially in younger patients.The FeAST data show that it should be possible to test definitively whether controlling iron levels may reduce disease risk and additional research is needed to further define ferrotoxic diseases, stratifying risk reduction with intervention, and clarify mechanisms, especially in younger patients. Zacharski et al. JAMA; 2007 Feb. 297(6):