Clinical validation of the PrecivityAD2 blood test: A mass spectrometry-based test with algorithm combining %p-tau217 and Aß42/40 ratio to identify presence of brain amyloid.

BACKGROUND: With the availability of disease-modifying therapies for Alzheimer's disease (AD), it is important for clinicians to have tests to aid in AD diagnosis, especially when the presence of amyloid pathology is a criterion for receiving treatment. METHODS: High-throughput, mass spectrometry-based assays were used to measure %p-tau217 and amyloid beta (Aß)42/40 ratio in blood samples from 583 individuals with suspected AD (53% positron emission tomography [PET] positive by Centiloid > 25). An algorithm (PrecivityAD2 test) was developed using these plasma biomarkers to identify brain amyloidosis by PET. RESULTS: The area under the receiver operating characteristic curve (AUC-ROC) for %p-tau217 (0.94) was statistically significantly higher than that for p-tau217 concentration (0.91). The AUC-ROC for the PrecivityAD2 test output, the Amyloid Probability Score 2, was 0.94, yielding 88% agreement with amyloid PET. Diagnostic performance of the APS2 was similar by ethnicity, sex, age, and apoE4 status. DISCUSSION: The PrecivityAD2 blood test showed strong clinical validity, with excellent agreement with brain amyloidosis by PET.

Assessment of a Plasma Amyloid Probability Score to Estimate Amyloid Positron Emission Tomography Findings Among Adults With Cognitive Impairment.

Importance: The diagnostic evaluation for Alzheimer disease may be improved by a blood-based diagnostic test identifying presence of brain amyloid plaque pathology. Objective: To determine the clinical performance associated with a diagnostic algorithm incorporating plasma amyloid-ß (Aß) 42:40 ratio, patient age, and apoE proteotype to identify brain amyloid status. Design, Setting, and Participants: This cohort study includes analysis from 2 independent cross-sectional cohort studies: the discovery cohort of the Plasma Test for Amyloidosis Risk Screening (PARIS) study, a prospective add-on to the Imaging Dementia-Evidence for Amyloid Scanning study, including 249 patients from 2018 to 2019, and MissionAD, a dataset of 437 biobanked patient samples obtained at screenings during 2016 to 2019. Data were analyzed from May to November 2020. Exposures: Amyloid detected in blood and by positron emission tomography (PET) imaging. Main Outcomes and Measures: The main outcome was the diagnostic performance of plasma Aß42:40 ratio, together with apoE proteotype and age, for identifying amyloid PET status, assessed by accuracy, sensitivity, specificity, and area under the receiver operating characteristic curve (AUC). Results: All 686 participants (mean [SD] age 73.2 [6.3] years; 368 [53.6%] men; 378 participants [55.1%] with amyloid PET findings) had symptoms of mild cognitive impairment or mild dementia. The AUC of plasma Aß42:40 ratio for PARIS was 0.79 (95% CI, 0.73-0.85) and 0.86 (95% CI, 0.82-0.89) for MissionAD. Ratio cutoffs for Aß42:40 based on the Youden index were similar between cohorts (PARIS: 0.089; MissionAD: 0.092). A logistic regression model (LRM) incorporating Aß42:40 ratio, apoE proteotype, and age improved diagnostic performance within each cohort (PARIS: AUC, 0.86 [95% CI, 0.81-0.91]; MissionAD: AUC, 0.89 [95% CI, 0.86-0.92]), and overall accuracy was 78% (95% CI, 72%-83%) for PARIS and 83% (95% CI, 79%-86%) for MissionAD. The model developed on the prospectively collected samples from PARIS performed well on the MissionAD samples (AUC, 0.88 [95% CI, 0.84-0.91]; accuracy, 78% [95% CI, 74%-82%]). Training the LRM on combined cohorts yielded an AUC of 0.88 (95% CI, 0.85-0.91) and accuracy of 81% (95% CI, 78%-84%). The output of this LRM is the Amyloid Probability Score (APS). For clinical use, 2 APS cutoff values were established yielding 3 categories, with low, intermediate, and high likelihood of brain amyloid plaque pathology. Conclusions and Relevance: These findings suggest that this blood biomarker test could allow for distinguishing individuals with brain amyloid-positive PET findings from individuals with amyloid-negative PET findings and serve as an aid for Alzheimer disease diagnosis.

The PrecivityAD™ test: Accurate and reliable LC-MS/MS assays for quantifying plasma amyloid beta 40 and 42 and apolipoprotein E proteotype for the assessment of brain amyloidosis.

BACKGROUND: There is an unmet need for an accessible, less invasive, cost-effective method to facilitate clinical trial enrollment and aid in clinical Alzheimer's disease (AD) diagnosis. APOE genotype affects the clearance and deposition of amyloid-beta (Aß) with APOE4 carriers having increased risk while APOE2 alleles appear to be protective. Lower plasma Aß42/40 correlates with brain amyloidosis. In response, C2N has developed the PrecivityAD™ test; plasma LC-MS/MS assays for Aß isoform quantitation and qualitative APOE isoform-specific proteotyping. METHODS: In accord with CLIA standards, we developed and validated assay performance: precision, accuracy, linearity, limit of detection (LoD), interferences. RESULTS: Within-day precision varied from 1.5-3.0% (Aß40) and 2.5-8.4% (Aß42). Total (within-lab) variability was 2.7-7.7% (Aß40) and 3.1-9.5% (Aß42). Aß40 quantitation was linear from 10 to 1780 pg/mL; Aß42 was linear from 2 to 254 pg/mL. LoD was 11 and 2 pg/mL for Aß40 and Aß42, respectively. APOE proteotypes were 100% concordant with genotype, while LoD (fM) was much lower than APOE concentrations observed in plasma (mM). CONCLUSIONS: The PrecivityAD™ assays are precise, accurate, sensitive, and linear over a wide analytical range, free from significant interferences, and suitable for use in the clinical laboratory.

Framingham risk equation underestimates subclinical atherosclerosis risk in asymptomatic women.

BACKGROUND: Coronary heart disease (CHD) is the leading cause of death among American women. Currently, global risk assessment derived by Framingham risk equation (FRE) is used to identify women at increased risk for CHD. Electron-beam computed tomography (EBCT) derived coronary artery calcium (CAC) scores are validated markers for future CHD events among asymptomatic individuals. However, the adequacy of FRE for identifying asymptomatic women with CAC is unknown. METHODS AND RESULTS: We studied 2447 consecutive non-diabetic asymptomatic females (55 +/- 10 years). Based upon FRE, 90% were classified as low-risk (FRE < or = 9% 10-year risk of hard CHD events), 10% intermediate-risk (10-20%), and none were considered as high-risk (> 20%). Coronary artery calcium was present in 33%, whereas CAC > or = 100 and CAC > or = 400 were seen in 10 and 3% of women, respectively. Overall, 20% of women had age-gender derived > or = 75th percentile CAC. According to FRE, the majority (84%) of women with significant CAC > or = 75th percentile were classified as low-risk. Approximately half (45%) of low-risk women with > or = 2 CHD risk factors and a family history of premature CHD had significant CAC. CONCLUSION: Framingham risk equation frequently classifies women as being low-risk, even in the presence of significant CAC. Determination of CAC may provide incremental value to FRE in identifying asymptomatic women who will benefit from targeted preventative measures.

Difference in atherosclerosis burden in different nations and continents assessed by coronary artery calcium.

We utilized coronary artery calcium scores (CACS) to assess differences in atherosclerosis burden between asymptomatic White populations living in continents with different cardiovascular disease rates. The similarities in the genetic pool between Brazilian and Portuguese Caucasian subjects offered an opportunity to assess the influence of environmental factors on the development of atherosclerosis. We reviewed CACS data from 17,563 individuals (12,378 men and 5169 women) collected in the USA (74% of the subjects), Brazil (15% of the subjects) and Portugal (11% of the subjects). CACS was absent in 80 and 88% of Portuguese men and women, compared with 46 and 62% and 33 and 59% of Brazilian and US counterparts (p<0.0001). Although the US subjects showed the lowest prevalence of risk factors they had a higher median (interquartile range) CACS than the Brazilian and the Portuguese cohorts: 4 (0;87), 1 (0;68) and 0 (0;0), respectively (p<0.0001). After adjusting for differences in age and cardiovascular risk factors, US men showed higher relative risk ratios of having any CACS than either Brazilian or Portuguese men. Brazilian and US women did not differ as far as risk of CACS although they demonstrated a greater risk than Portuguese women. In this study, significant differences in CACS were detected among three nations in different continents. The CACS differences paralleled the respective cardiovascular mortality rates.

The influence of health status, age, and race on screening mammography in elderly women.

BACKGROUND: Screening mammography is controversial for elderly women because of an absence of efficacy data. Decisions to screen are based on individualized assessment of risks and benefits. Our objective was to determine how screening mammography varies by age and race when adjusted for propensity to die. METHODS: In a retrospective cohort study, rates of screening mammogram performed in 2000-2001 based on claims, adjusted for propensity to die in 2000, were determined for a nationally representative 5% random sample of female fee-for-service Medicare beneficiaries 65 years and older in (N = 722,310). RESULTS: The overall rate of screening was 39%. When stratified into quintiles by propensity to die, 2-year rates ranged from 61% in the lowest-risk group to 5% in the highest-risk group. In analyses stratified by age and adjusted for propensity to die, 42% of women aged 65 to 69 years were screened, declining to 26% of women 85 years and older (P<.001). Adjusted screening rates for white women, black women, and women of other races were 40%, 30%, and 25%, respectively (P<.001). Thus, among women with similar health status, the youngest women were 1.61 times more likely to be screened compared with the oldest; compared with black women and women of other races, white women were 1.38 and 1.60 times, respectively, more likely to be screened. CONCLUSIONS: Decisions to screen for breast cancer are related not only to health status but also to age and race. Underuse and overuse of screening mammography likely occurs owing to age- and race-associated decision making. Assessment of life expectancy may more accurately identify women who could benefit from screening.

Relation of family history of premature coronary heart disease and metabolic risk factors to risk of coronary arterial calcium in asymptomatic subjects.

We studied 6,141 consecutive, asymptomatic, nondiabetic patients who underwent electron beam tomography and explored the interaction between metabolic risk factors (RFs) and premature family history (FH) of coronary heart disease (CHD) in predicting the presence and severity of coronary arterial calcium (CAC). In the presence of >2 metabolic RFs, patients with a positive FH of premature CHD had a significantly higher prevalence of any CAC, CAC >/=100, and CAC >/=75th age-gender percentile than those without a FH of CHD. Our study demonstrated that a familial propensity to subclinical atherosclerosis interacts with the presence of >/=2 metabolic RFs, magnifying the risks for those exposed to both.

Underlying risk factors incrementally add to the standard risk estimate in detecting subclinical atherosclerosis in low- and intermediate-risk middle-aged asymptomatic individuals.

BACKGROUND: Traditional risk factors predict the 10-year risk of developing coronary heart disease (CHD). Underlying risk factors like physical inactivity, obesity (BMI >30 kg/m2), and family history of premature CHD are independently associated with CHD. High burden of coronary artery calcification (CAC) on electron beam tomography (EBT) is a CHD risk equivalent. GOALS: To determine the association between CAC and traditional risk assessment and whether the presence of added underlying risk factors is associated with advanced CAC in low- and intermediate-risk individuals. METHODS: After excluding patients with CHD, we studied 8549 asymptomatic individuals referred for EBT for cardiac risk assessment. Traditional myocardial infarction risk score was estimated according to Framingham criteria, and individuals were divided into 3 groups: low-risk (< or =9% MI risk over the next 10 years), intermediate-risk (10%-20% risk), and high-risk (>20 % risk). Advanced CAC was defined as a calcium score > or =75th percentile based on sex and age. RESULTS: The prevalence of advanced CAC was 20% in low-risk, 27% in intermediate-risk, and 31% in high-risk individuals (P < .001). Underlying risk factors were incrementally associated with advanced CAC in low- and intermediate-risk individuals (P < .001). A receiver operating characteristic curve analysis revealed that addition of underlying risk factors to traditional risk assessment increased the area under the curve significantly from 0.56 to 0.62 (P < .001). CONCLUSION: A substantial proportion of low- and intermediate-risk individuals have advanced CAC. Adding family history, obesity, and physical inactivity to traditional risk assessment improves prediction of advanced CAC, which may lead these individuals to be treated more aggressively at an earlier age.

Coronary artery calcification and family history of premature coronary heart disease: sibling history is more strongly associated than parental history.

BACKGROUND: The objective of the study was to assess the association of a family history (FH) of premature coronary heart disease (CHD) with coronary artery calcification (CAC) in asymptomatic individuals and to compare the effects of sibling or parental FH on the risk of subclinical atherosclerosis. METHODS AND RESULTS: CAC by electron beam tomography was performed in 8549 asymptomatic individuals (69% men; mean age, 52+/-9 years). The prevalence and odds of any CAC and extent of CAC stratified according to FH of premature CHD were determined. Those with (1) no FH of CHD, (2) FH of premature CHD in parents, or (3) FH in siblings had a prevalence of CAC of 55%, 64%, and 78% (P<0.0001) among men and 27%, 36%, and 56% (P<0.0001) among women, respectively. The multivariate regression analysis demonstrated that the odds ratio (95% confidence interval) for the presence of CAC was 1.3 (1.1 to 1.6) among those with positive FH of premature CHD in parents only, 2.3 (1.7 to 3.1) and 2.5 (1.8 to 3.3) among those in siblings and a combined FH compared with those without FH of CHD in men, respectively. Among women, the corresponding odds ratios were 1.3 (1.0 to 1.8), 2.3 (1.7 to 3.6), and 1.9 (1.3 to 3.1), respectively. A similar trend was observed in the association of FH of premature CHD with increasing CAC scores. CONCLUSIONS: Our study demonstrates a highly significant association between FH of premature CHD and the presence and extent of CAC. Furthermore, within the limits of self-reporting of family history, our findings suggest that a sibling history is more strongly associated with subclinical coronary atherosclerosis than a parental history of premature CHD.

Relation of degree of physical activity to coronary artery calcium score in asymptomatic individuals with multiple metabolic risk factors.

Lack of physical activity (PA) increases risk of coronary heart disease. Metabolic risk factors increase the risk of coronary heart disease and development of advanced coronary artery calcium (CAC). We hypothesized that, in a population with multiple metabolic risk factors (> or =2), the degree of PA would be inversely associated with the degree and prevalence of CAC. After excluding subjects who had known diabetes and coronary heart disease, we studied 779 asymptomatic patients referred for electron beam tomography. All patients had > or =2 of the following metabolic risk factors: blood pressure > 130/85 mm Hg, serum triglycerides >150 mg/dl, serum high-density lipoprotein < 40 and < 50 mg/dl in men and women, respectively, and body mass index >30 kg/m(2). Advanced CAC was defined as a calcium score >75th percentile based on gender and age. In the study, 37% patients were sedentary, 26% engaged in moderate-duration (< 30 minutes 1 to 2 times/week) PA, and 37% engaged in long-duration (> or =30 minutes > or =3 times/week) PA. The median CAC scores were 24 (sedentary), 18 (moderate PA), and 11 (long-duration PA; p <0.002). Advanced CAC was prevalent in 26% of sedentary patients, 24% of patients who performed moderate PA, and 16% of patients who engaged in long-duration PA (p <0.05). On logistic regression analysis, long-duration PA had an independent inverse association with advanced CAC. Thus, asymptomatic patients who have > or =2 metabolic risk factors and who regularly engage in long-duration PA have a lower prevalence of CAC than do those who are sedentary or participate in moderate-duration PA.

Coronary artery calcium volume scores on electron beam tomography in 12,936 asymptomatic adults.

We developed age- and gender-specific normative tables of calcium volume scores by using data from 12,936 asymptomatic patients who underwent electron beam tomographic scanning and compared the volume with the Agatston scores obtained in the same subjects. The 2 scores increased as the number of atherosclerotic risk factors increased. The volume scores were statistically smaller than the Agatston scores at the upper quartile level.

Prospective evaluation of the relationship between platelet-leukocyte conjugate formation and recurrent myocardial ischemia in patients with acute coronary syndromes.

Platelet-leukocyte conjugates are increased in patients with coronary artery disease but the relationship between conjugate formation and myocardial ischemic outcome is unknown. We prospectively evaluated the relationship between conjugate formation and the development of recurrent myocardial ischemia in patients with acute coronary syndromes (ACS). Platelet-leukocyte conjugate formation (induced by thrombin receptor activating peptide (TRAP)) and platelet aggregation (induced by ADP and arachidonic acid) were assessed in 30 patients with unstable angina or non-ST elevation myocardial infarction. All patients were treated with beta-blockers, aspirin, heparin, and GPIIb-IIIa antagonists and were followed for in-hospital recurrent myocardial ischemia. Troponin I and C-reactive protein (CRP) were also measured. Seven patients (23%) experienced recurrent ischemia. Platelet-neutrophil conjugates were greater in ischemic patients (59 +/- 9 and 36 +/- 4%, P = 0.007, for + ischemia and -ischemia, respectively). Platelet aggregation did not differ between ischemic and nonischemic patients, and there was no significant relationship between aspirin resistance and ischemic outcome. Troponin I was greater in patients who developed recurrent ischemia (3.04 +/- 1.73 vs. 0.70 +/- 0.21 ng/ml, P = 0.03, for +ischemia and -ischemia, respectively) but CRP was not. TRAP-induced platelet-neutrophil conjugate formation was an independent predictor of ischemic outcome (OR 1.07, 95% CI 1.00-1.15, for each 1% increase in conjugate formation). Receiver operator characteristic analysis showed platelet-neutrophil conjugates to have good ability to discriminate between ischemic and nonischemic patients (AUC of 0.84, P < 0.05). TRAP induced platelet-neutrophil conjugate formation is related to in vivo ischemic risk in ACS patients.

Reducing global risk for cardiovascular disease: using lifestyle changes and pharmacotherapy.

Cardiovascular disease (CVD) is the leading cause of death and disability in industrialized societies, due in large part to the lack of a comprehensive approach to control the risk factors for atherosclerosis. One strategy for reducing an individual's global CVD risk relies on a targeted approach that modifies each of the major independent risk factors prevalent in both symptomatic (secondary prevention) and asymptomatic (primary prevention) patients. These interventions include lipid lowering, smoking cessation, blood pressure control, glycemic control, regular exercise, and the use of various medications. This review offers an evidence-based strategy toward reducing an individual's global risk for CVD by addressing the modifiable, major independent risk factors.