Discovery and validation of cell cycle arrest biomarkers in human acute kidney injury.

INTRODUCTION: Acute kidney injury (AKI) can evolve quickly and clinical measures of function often fail to detect AKI at a time when interventions are likely to provide benefit. Identifying early markers of kidney damage has been difficult due to the complex nature of human AKI, in which multiple etiologies exist. The objective of this study was to identify and validate novel biomarkers of AKI. METHODS: We performed two multicenter observational studies in critically ill patients at risk for AKI - discovery and validation. The top two markers from discovery were validated in a second study (Sapphire) and compared to a number of previously described biomarkers. In the discovery phase, we enrolled 522 adults in three distinct cohorts including patients with sepsis, shock, major surgery, and trauma and examined over 300 markers. In the Sapphire validation study, we enrolled 744 adult subjects with critical illness and without evidence of AKI at enrollment; the final analysis cohort was a heterogeneous sample of 728 critically ill patients. The primary endpoint was moderate to severe AKI (KDIGO stage 2 to 3) within 12 hours of sample collection. RESULTS: Moderate to severe AKI occurred in 14% of Sapphire subjects. The two top biomarkers from discovery were validated. Urine insulin-like growth factor-binding protein 7 (IGFBP7) and tissue inhibitor of metalloproteinases-2 (TIMP-2), both inducers of G1 cell cycle arrest, a key mechanism implicated in AKI, together demonstrated an AUC of 0.80 (0.76 and 0.79 alone). Urine [TIMP-2]·[IGFBP7] was significantly superior to all previously described markers of AKI (P <0.002), none of which achieved an AUC >0.72. Furthermore, [TIMP-2]·[IGFBP7] significantly improved risk stratification when added to a nine-variable clinical model when analyzed using Cox proportional hazards model, generalized estimating equation, integrated discrimination improvement or net reclassification improvement. Finally, in sensitivity analyses [TIMP-2]·[IGFBP7] remained significant and superior to all other markers regardless of changes in reference creatinine method. CONCLUSIONS: Two novel markers for AKI have been identified and validated in independent multicenter cohorts. Both markers are superior to existing markers, provide additional information over clinical variables and add mechanistic insight into AKI. TRIAL REGISTRATION: ClinicalTrials.gov number NCT01209169.

The use of B-type natriuretic peptide to assess volume status in patients with end-stage renal disease.

BACKGROUND: Although B-type natriuretic peptide (BNP) levels correlate with volume overload in congestive heart failure, its usefulness in patients with renal dysfunction has been questioned. A simple test to estimate volume overload and assist in the management of dry weight in hemodialysis (HD) patients would be useful. METHODS: Thirty-nine patients--aged 64 +/- 2 years (mean +/- SEM), male-female ratio of 37:2--undergoing HD thrice weekly for at least 30 days were studied. Samples were collected at the start and end of each of 3 consecutive HD sessions. Pre- and postsession weights and blood pressures were recorded. Left ventricular ejection fractions were obtained from echocardiograms performed within 1 year of enrollment. The first session was the dialysis session after a 72-hour interdialytic period, whereas the second and third sessions were after a 48-hour period. Plasma volume changes were measured in a subset of 13 patients. RESULTS: Pre- and postdialysis BNP levels for each of the 3 sessions were 434 and 343 pg/mL, 347 and 231 pg/mL, and 249 and 202 pg/mL, respectively. The values for body weights were 82.6 +/- 3.6 and 78.6 +/- 3.5 kg, 81.5 +/- 3.6 and 78.2 +/- 3.5 kg, and 81.5 +/- 3.46 and 78.3 +/- 3.5 kg, respectively. The values of mean systolic blood pressures were 150 +/- 4 and 134 +/- 3 mm Hg, 142 +/- 4 and 134 +/- 4 mm Hg, and 142 +/- 4 and 131 +/- 4 mm Hg, respectively. The values for mean diastolic blood pressures were 81 +/- 2.5 and 70 +/- 2.4 mm Hg, 74 +/- 2.4 and 72.1 +/- 2.2 mm Hg, and 76 +/- 2.9 and 72 +/- 2.9 mm Hg, respectively. There was no correlation between changes in intradialytic BNP values and other measured parameters. Plasma volume changed minimally during dialysis. CONCLUSIONS: Values of BNP are elevated in patients with end-stage renal disease and decline after each dialysis session. Over the course of a week, BNP levels gradually declined irrespective of changes in weight or blood pressure. The lack of correlation between changes in BNP and changes in measured clinical parameters is partly explained by a lack of a significant change in plasma volume. The highest BNP values were seen in patients with systolic dysfunction.

Hemodialysis access assessment with intravascular ultrasound.

Reliable identification and treatment of specific hemodialysis access complications may improve access patency and result in significant cost reduction. Angiography is the gold standard for the evaluation of vascular access; however, it has significant limitations. Intravascular ultrasound (IVUS) is a relatively new technique capable of detecting subtle vascular abnormalities. To investigate the safety, feasibility, and accuracy of IVUS imaging to detect hemodialysis access complications, including stenoses, graft deterioration, and thrombus, we performed 31 IVUS imaging studies in 22 hemodialysis patients. Nineteen studies were performed in the dialysis unit, and 12 studies in the angiography suite. The IVUS catheter was inserted into the graft through the access used for hemodialysis. Findings of 21 studies (17 patients) imaged on the same day by both angiography and IVUS were compared. Grafts and vessels were successfully imaged using IVUS in 29 of 31 studies. There were no adverse effects caused by IVUS. Angiography assessed 17 of 54 vessel segments as normal versus 9 of 54 segments by IVUS (P < 0.001). Angiography detected lesions in 25 segments as opposed to 33 segments by IVUS (P < 0.001). A thrombus was detected in 32 of 54 vessel segments by IVUS, but in only 1 of 54 segments by angiography (P < 0.001). In conclusion, IVUS imaging is feasible and safe to assess hemodialysis access in the angiographic suite and dialysis unit. IVUS detected more vascular abnormalities than angiography. IVUS may be a useful independent imaging and screening modality in the assessment of dialysis access complications, which may help increase graft patency and reduce cost.

41Ca and accelerator mass spectrometry to monitor calcium metabolism in end stage renal disease patients.

BACKGROUND: Monitoring bone resorption with measurements of bone density and biochemical markers is indirect. We hypothesized that bone resorption can be studied directly by serial measurements of the ratio (41)Ca/Ca in serum after in vivo labeling of calcium pools with (41)Ca. We report the preparation of an intravenous (41)Ca dose suitable for humans, an analytical method for determining (41)Ca/Ca isotope ratios in biological samples, and studies in human volunteers. METHODS: (41)Ca was formulated and aliquoted into individual vials, and to the extent possible, the (41)Ca doses were tested according to US Pharmacopeia (USP) guidelines. A 10 nCi dose of (41)Ca was administered intravenously to 4 end stage renal disease (ESRD) patients on hemodialysis and 4 healthy control individuals. Distribution kinetics were determined over 168 days. Calcium was isolated with 3 precipitation steps and a cation-exchange column, and (41)Ca/Ca ratios in serum were then measured by accelerator mass spectrometry. RESULTS: The dosing solution was chemically and radiologically pure, contained <0.1 endotoxin unit/mL, and passed USP sterility tests. Quantification of (41)Ca/Ca ratios was linear from 6 x 10(-14) to 9.1 x 10(-10). The run-to-run imprecision (as CV) of the method was 4% at 4.6 x 10(-11) and 6% at 9.1 x 10(-10). The area under the curve of (41)Ca in the central compartment vs time was significantly less for ESRD patients than for controls (P < 0.005). CONCLUSIONS: Isotope ratios spanning 5 orders of magnitude can be measured by accelerator mass spectrometry with excellent precision in the range observed in samples collected from patients who have received 10 nCi of (41)Ca. The (41)Ca at this dose caused no adverse effects in 8 volunteers. This is the first report of the use of (41)Ca to monitor differences in bone turnover between healthy individuals and ESRD patients.