Transcriptional profiling in coronary artery disease: indications for novel markers of coronary collateralization.

BACKGROUND: The development of collateral circulation plays an important role in protecting tissues from ischemic damage, and its stimulation has emerged as one of principal approaches to therapeutic angiogenesis. Clinical observations have documented substantial differences in the extent of collateralization among patients with coronary artery disease (CAD), with some individuals demonstrating marked abundance and others showing nearly complete absence of these vessels. Recent studies have suggested that circulating monocytes play a major role in collateral growth. The present study was undertaken to determine transcriptional profiles of circulating monocytes in CAD patients with different extents of collateral growth. METHODS AND RESULTS: Monocyte transcriptomes from CAD patients with and without collateral vessels were obtained by use of high-throughput expression profiling. Using a newly developed redundancy-based data mining method, we have identified a set of molecular markers characteristic of a "noncollateralgenic" phenotype. Moreover, we show that these transcriptional abnormalities are independent of the severity of CAD or any other known clinical parameter thought to affect collateral development and correlated with protein expression levels in monocytes and plasma. CONCLUSIONS: Monocyte transcription profiling identifies sets of patients with extensive versus poorly developed collateral circulation. Thus, genetic factors may heavily influence coronary collateral vessel growth in CAD and affect prognosis and response to therapeutic interventions.

Impact of mouse strain differences in innate hindlimb collateral vasculature.

OBJECTIVE: To assess the importance of genetic background for collateral artery development. METHODS AND RESULTS: C57BL/6, BALB/c and 129S2/Sv mice were studied after femoral artery ligation by laser Doppler imaging, visible light oximetry, time-of-flight-magnetic resonance imaging, and treadmill testing; C57BL/6 and BALB/c also underwent electron paramagnetic resonance (EPR) oximetry, x-ray angiography, and histology. C57BL/6 had the least initial distal ischemia and most complete recovery. BALB/c had the most severe initial ischemia and poorest recovery. BALB/c had some vasodilatory reserve in their ligated limbs not seen in the other strains at 3 weeks. By in vivo TOF-magnetic resonance angiography, C57BL/6 had larger preexistent and developed collaterals. By x-ray angiography, C57BL/6 had a higher collateral-dependent filling score and number of visible collaterals immediately after femoral ligation and a higher number of visible collaterals at 1 week but not at 4 weeks. EPR oximetry and histology revealed hypoxia and tissue damage in regions of collateral growth of BALB/c but not C57BL/6 mice. In C57BL/6 BrdUrd uptake in the thigh was limited to larger vessels and isolated perivascular cells. Proliferative activity in collateral arterioles was similar in both strains. CONCLUSIONS: Genetic differences in preexistent collateral vasculature can profoundly affect outcome and milieu for compensatory collateral artery growth after femoral artery occlusion.

Automated migration analysis based on cell texture: method & reliability.

BACKGROUND: In this paper, we present and validate a way to measure automatically the extent of cell migration based on automated examination of a series of digital photographs. It was designed specifically to identify the impact of Second Hand Smoke (SHS) on endothelial cell migration but has broader applications. The analysis has two stages: (1) preprocessing of image texture, and (2) migration analysis. RESULTS: The output is a graphic overlay that indicates the front lines of cell migration superimposed on each original image, with automated reporting of the distance traversed vs. time. Expert preference compares to manual placement of leading edge shows complete equivalence of automated vs. manual leading edge definition for cell migration measurement. CONCLUSION: Our method is indistinguishable from careful manual determinations of cell front lines, with the advantages of full automation, objectivity, and speed.

Detection of exercise-induced ischemia by changes in B-type natriuretic peptides.

OBJECTIVES: The purpose of this study was to examine the effect of exercise-induced ischemia on levels of B-type natriuretic peptide (BNP) and its inactive N-terminal fragment (NT-pro-BNP)and to determine whether measurement of these peptides can improve the diagnostic accuracy of exercise testing. BACKGROUND: The ability of exercise testing to detect coronary artery disease (CAD) is limited by modest sensitivity and specificity. B-type natriuretic peptides (NT-pro-BNP and BNP) are released by ventricular myocytes in response to wall stress. We hypothesized that exercise-induced ischemia results in increased wall stress and triggers release of NT-pro-BNP and BNP. METHODS: A total of 74 patients with known CAD, normal left ventricular function, and normal resting levels of NT-pro-BNP and BNP who were referred for exercise testing with radionuclide imaging, and 21 healthy volunteers, were enrolled. Blood was drawn before and after maximal exercise and analyzed for NT-pro-BNP and BNP. RESULTS: Of the patients with CAD, 40 had ischemia on perfusion images and 34 did not. Median post-exercise increases in NT-pro-BNP and BNP (DeltaNT-pro-BNP and DeltaBNP) were approximately four-fold higher in the ischemic group than in the nonischemic group (DeltaNT-pro-BNP 14.5 vs. 4 pg/ml, p < 0.0001; DeltaBNP 36.5 vs. 7.5 pg/ml, p < 0.0001). In volunteers, median DeltaNT-pro-BNP was almost identical to that of the nonischemic patient group. At equal specificity to the electrocardiogram (ECG) (58.8%), the sensitivities of DeltaNT-pro-BNP and DeltaBNP for detecting ischemia were 90% and 80%, respectively; in contrast, the sensitivity of the exercise ECG was 37.5%. CONCLUSIONS: Measurement of exercise-induced increases in BNPs more than doubles the sensitivity of the exercise test for detecting ischemia with no loss of specificity.

Magnetic resonance imaging demonstrates improved regional systolic wall motion and thickening and myocardial perfusion of myocardial territories treated by laser myocardial revascularization.

OBJECTIVES: This study was designed to investigate the use of magnetic resonance (MR) functional and perfusion imaging to evaluate laser myocardial revascularization (LMR). BACKGROUND: Most clinical studies of LMR have shown improvements in angina class and exercise capacity, with minimal or absent improvements in myocardial perfusion and function. METHODS: Fifteen patients who underwent percutaneous Biosense-guided holmium:yttrium aluminum garnet LMR to areas of viable but ischemic myocardium were followed clinically and underwent functional and perfusion MRI at baseline, 30 days and 6 months. RESULTS: The mean age was 64 +/- 11 years; four patients were women. The ejection fraction was 47.4 +/- 14.0%. Angina class at baseline was 3.4 +/- 0.6 and improved to 2.5 +/- 1.4 at six months (p = 0.054). Exercise time at baseline was 298 +/- 97 s and increased to 350 +/- 95 s at 30 days and 365 +/- 79 s at six months, p = 0.04. There were no significant changes in nuclear perfusion scans. Although MR determined that resting radial motion and thickening of the target wall were significantly less than normal at baseline (p < 0.001), they improved significantly during follow-up (wall thickening: baseline, 30.6 +/- 11.7%; day 30, 41.2 +/- 13.3% and day 180, 44.2 +/- 11.9%, p = 0.01). The size of the underperfused myocardial area was 14.5 +/- 5.4% at baseline and was reduced to 6.3 +/- 2.8% at 30 days and 7.7 +/- 3.7% at 6 months (p < 0.001). CONCLUSIONS: This small phase I, open-label, uncontrolled study of MR functional and perfusion imaging in patients undergoing Biosense-guided LMR suggests a beneficial effect of this treatment strategy on myocardial function and perfusion. The efficacy of Biosense-guided LMR is being evaluated in a large phase II, randomized, blinded placebo-controlled trial with an MRI substudy (DIRECT).

Medical imaging techniques in the evaluation of strategies for therapeutic angiogenesis.

Recent advancements in our understanding of the basic biology of angiogenesis have prompted a focus on practical applications, both in cardiovascular disease and in oncology. The focus on practical applications has stimulated development of novel noninvasive tools that provide serial assessment of ongoing vessel growth in vivo. Nuclear imaging (SPECT, PET) and x-ray angiography have been used to assess changes in perfusion and anatomic appearance, respectively, after induced neovascular development. New MRI techniques provide the ability to identify early changes in vivo that are more sensitive to detection of the effects of new vessel growth than x-ray angiography or nuclear imaging. These new MRI techniques include measurement of blood delivery to the myocardium, development of intramyocardial vasculature, and incremental changes in regional myocardial contractile function. With the combination of methods now available, we expect to be able to track key steps of angiogenesis in vivo and to assess the efficacy of angiogenic therapies. These new imaging capabilities offer crucial information which we hope will hasten the identification and deployment of effective pharmaceutical therapies as an adjunct or alternative to invasive treatments of ischemic disease by targeted stimulation of angiogenesis, and of cancer, by targeted inhibition of angiogenesis.

Adenoviral PR39 improves blood flow and myocardial function in a pig model of chronic myocardial ischemia by enhancing collateral formation.

Angiogenic therapy with individual growth factors or "master switch" genes is being evaluated for treatment of advanced coronary artery disease. In this study, we investigated the efficacy and mechanism of PR39, a gene capable of activating VEGF and fibroblast growth factor (FGF)-2-dependent pathways. PR39 enhances hypoxia-inducible factor-1alpha (HIF-1alpha)-dependent gene expression by selectively inhibiting proteasome degradation of this transcription factor. In addition, PR39 also stimulates expression of the FGF receptors (FGFR)-1 and syndecan-4. In a pig model of chronic myocardial ischemia, we used angiography, MRI, and microsphere regional blood flow to evaluate the efficacy of intramyocardial adenoviral protein arginine-rich peptide (Ad-PR39) injections. Ad-PR39 improved collateral scores, regional perfusion, and regional function in a dose-dependent manner. Local VEGF, VEGFR-1, VEGFR-2, syndecan, and FGFR-1 levels were 16-75% upregulated after Ad-PR39 injections as assessed by real-time PCR, suggesting upregulation of VEGF and FGF pathways. PR39 is an angiogenic peptide that improves perfusion and function of ischemic myocardium, at least in part, through collateral formation. The dual mechanism, i.e., stimulation of HIF-1alpha and FGF receptor expression, likely accounts for the functional benefits of PR39.

Arteriogenesis: noninvasive quantification with multi-detector row CT angiography and three-dimensional volume rendering in rodents.

PURPOSE: To evaluate two-dimensional (2D) multi-detector row computed tomographic (CT) angiography and three-dimensional (3D) volume rendering for depiction of patterns of arterial growth and quantification of blood vessel density and volume. MATERIALS AND METHODS: The institutional animal care and use committee approved this study. The right femoral artery and its branches were ligated and excised in 16 inbred Lewis rats; animals were randomly assigned to receive 70 microL Dulbecco's modified Eagle's medium (DMEM) or 1.5 x 10(7) bone marrow-derived mononuclear cells (BMC) from isogenic donor rats in 70 microL DMEM. At 2 weeks, CT angiography was performed with injection of 0.45 mL barium sulfate suspension at 0.7 mL/min, followed by silver staining. Number of blood vessels, area, mean area, volume, and blood vessel size distribution derived from digitally subtracted 2D CT angiographic sections were quantified; 3D images were reconstructed. Two-way analysis of variance and paired and unpaired Student t tests were performed. RESULTS: CT angiography showed two patterns of arterial growth: collateral arterial formation and branching arteriogenesis. Two-way analysis of variance indicated that differences within subjects (ischemic vs nonischemic legs) and between subjects (BMC vs DMEM treatment) were significant for total blood vessel area, total blood vessel volume, and mean of blood vessel area (P < .001). In the BMC group, there were significantly more arteries (mean, 241.6 +/- 77.0 [standard deviation] vs 196.4 +/- 75.2, P = .028), but mean cross-sectional area of these arteries was smaller in ischemic versus nonischemic legs (5.4 mm(2) +/- 1.2 vs 6.8 mm(2) +/- 1.3, P = .006). Total arterial area and volume did not differ significantly between ischemic and nonischemic legs. CONCLUSION: BMC injection had a substantial effect on arteriogenesis, with normalization of total arterial area and volume in the BMC group; this effect was successfully depicted.

Novel double contrast MRI technique for intramyocardial detection of percutaneously transplanted autologous cells.

Bone marrow cells (BMC) labeled with iron particles can be injected into the heart and detected with MRI. Improvement in conspicuity of labelled cells would be advantageous. This study examined if double contrast with iron oxide and Gd-DTPA enhances cell MRI after transvascular transplantation in myocardial infarction. Ten pigs with week-old myocardial infarction had transvascular peri-infarct delivery of microspheres alone (Group I, n = 3) or mixed with iron-labeled BMCs (Group II, n = 7). Gradient-echo MRI before and 1 min after systemic Gd-DTPA administration produced regions of interest with hypoenhancement that were compared to contralateral regions for contrast-to-noise (CNR) and signal-to-noise (SNR) ratios. All hearts were harvested for gross and microscopic analysis. Areas of focal hypoenhancement corresponding to the BMCs were detected in the myocardium in Group II. Early after administration of Gd-DTPA CNR increased from 17.58 +/- 8.5 to 27.25 +/- 15.8 (P < 0.05) and SNR from 24.87 +/- 9.6 to 35.08 +/- 15.5 (P < 0.05). There was no hypoenhancement in Group I. Tissue examination confirmed presence of iron-containing cells and microspheres in corresponding segments of the heart. The distribution of microspheres was similar between the groups. Double contrast with cellular iron and Gd-DTPA in surrounding myocardium resulted in improved cell localization by MRI.

An extensible framework for sharing clinical guidelines and services.

Accurate and descriptive information from clinical studies guides improvements in health care. Clinical guidelines established by authoritative medical organizations provide such information in a standard form for medical professionals' reference. Previous work on electronically sharing clinical guidelines focuses on the idea of building unified clinical terminologies and sharing resources through centralized data repositories. In this paper we propose a novel five-layer framework called the Extensible Clinical Guidelines and Services Sharing Architecture (ECGSSA). This framework provides for clinical guideline sharing among autonomous service providers over a distributed architecture. Requests for exchange of guidelines are disseminated through Web Services through a registry mechanism. Currently we have adopted the Guideline Interchange Format (GLIF) from InterMed as the representation format and use the Open Grid Services Architecture (OGSA) to attain virtual organization of shared guideline and service resources. This approach will allow more flexibility for medical professionals to exchange their practice guidelines in an effort to improve quality of health care. Also, it extends the possibility of solving clinic-related computational problems through collaborative sharing of analytical services. A sample scenario is presented to explain the application of ECGSSA in distributed task assignment and service matching in medical image processing.

BP-neural network based-characterization of electrographic magnetohydrodynamic signals in MR.

Electrocardiographs (ECG) signal collected during magnetic resonance (MR) imaging is affected by signal artifact because magnetic fields produce competing signals, from moving conductors in the large vessels. That is called the magnetohydrodynamic effect, which makes it difficult to recognize ST-T changes from ECG signal collected in a magnetic field (MRI). Resolving that problem is important both for accurate triggering (elimination of false triggers from tall peaked T waves) and for monitoring (identifying if or when patient develops ischemia or myocardial injury). This work describes an algorithm based on neural network that is designed to cancel this artifact for ECG signal acquired during MR imaging.