AZGP1 deficiency promotes angiogenesis in prostate cancer.

BACKGROUND: Loss of AZGP1 expression is a biomarker associated with progression to castration resistance, development of metastasis, and poor disease-specific survival in prostate cancer. However, high expression of AZGP1 cells in prostate cancer has been reported to increase proliferation and invasion. The exact role of AZGP1 in prostate cancer progression remains elusive. METHOD: AZGP1 knockout and overexpressing prostate cancer cells were generated using a lentiviral system. The effects of AZGP1 under- or over-expression in prostate cancer cells were evaluated by in vitro cell proliferation, migration, and invasion assays. Heterozygous AZGP1± mice were obtained from European Mouse Mutant Archive (EMMA), and prostate tissues from homozygous knockout male mice were collected at 2, 6 and 10 months for histological analysis. In vivo xenografts generated from AZGP1 under- or over-expressing prostate cancer cells were used to determine the role of AZGP1 in prostate cancer tumor growth, and subsequent proteomics analysis was conducted to elucidate the mechanisms of AZGP1 action in prostate cancer progression. AZGP1 expression and microvessel density were measured in human prostate cancer samples on a tissue microarray of 215 independent patient samples. RESULT: Neither the knockout nor overexpression of AZGP1 exhibited significant effects on prostate cancer cell proliferation, clonal growth, migration, or invasion in vitro. The prostates of AZGP1-/- mice initially appeared to have grossly normal morphology; however, we observed fibrosis in the periglandular stroma and higher blood vessel density in the mouse prostate by 6 months. In PC3 and DU145 mouse xenografts, over-expression of AZGP1 did not affect tumor growth. Instead, these tumors displayed decreased microvessel density compared to xenografts derived from PC3 and DU145 control cells, suggesting that AZGP1 functions to inhibit angiogenesis in prostate cancer. Proteomics profiling further indicated that, compared to control xenografts, AZGP1 overexpressing PC3 xenografts are enriched with angiogenesis pathway proteins, including YWHAZ, EPHA2, SERPINE1, and PDCD6, MMP9, GPX1, HSPB1, COL18A1, RNH1, and ANXA1. In vitro functional studies show that AZGP1 inhibits human umbilical vein endothelial cell proliferation, migration, tubular formation and branching. Additionally, tumor microarray analysis shows that AZGP1 expression is negatively correlated with blood vessel density in human prostate cancer tissues. CONCLUSION: AZGP1 is a negative regulator of angiogenesis, such that loss of AZGP1 promotes angiogenesis in prostate cancer. AZGP1 likely exerts heterotypical effects on cells in the tumor microenvironment, such as stromal and endothelial cells. This study sheds light on the anti-angiogenic characteristics of AZGP1 in the prostate and provides a rationale to target AZGP1 to inhibit prostate cancer progression.

Pulse Wave Imaging Coupled With Vector Flow Mapping: A Phantom, Simulation, and In Vivo Study.

Pulse wave imaging (PWI) is an ultrasound imaging modality that estimates the wall stiffness of an imaged arterial segment by tracking the pulse wave propagation. The aim of the present study is to integrate PWI with vector flow imaging, enabling simultaneous and co-localized mapping of vessel wall mechanical properties and 2-D flow patterns. Two vector flow imaging techniques were implemented using the PWI acquisition sequence: 1) multiangle vector Doppler and 2) a cross-correlation-based vector flow imaging (CC VFI) method. The two vector flow imaging techniques were evaluated in vitro using a vessel phantom with an embedded plaque, along with spatially registered fluid structure interaction (FSI) simulations with the same geometry and inlet flow as the phantom setup. The flow magnitude and vector direction obtained through simulations and phantom experiments were compared in a prestenotic and stenotic segment of the phantom and at five different time frames. In most comparisons, CC VFI provided significantly lower bias or precision than the vector Doppler method ( ) indicating better performance. In addition, the proposed technique was applied to the carotid arteries of nonatherosclerotic subjects of different ages to investigate the relationship between PWI-derived compliance of the arterial wall and flow velocity in vivo. Spearman's rank-order test revealed positive correlation between compliance and peak flow velocity magnitude ( rs = 0.90 and ), while significantly lower compliance ( ) and lower peak flow velocity magnitude ( ) were determined in older (54-73 y.o.) compared with young (24-32 y.o.) subjects. Finally, initial feasibility was shown in an atherosclerotic common carotid artery in vivo. The proposed imaging modality successfully provided information on blood flow patterns and arterial wall stiffness and is expected to provide additional insight in studying carotid artery biomechanics, as well as aid in carotid artery disease diagnosis and monitoring.

Atherosclerotic plaque mechanical characterization coupled with vector Doppler imaging in atherosclerotic carotid arteries in-vivo.

Methods used in clinical practice to diagnose and monitor atherosclerosis present limitations. Imaging the mechanical properties of the arterial wall has demonstrated the potential evaluate plaque vulnerability and assess the risk for stroke. Adaptive Pulse Wave Imaging (PWI) is a non-invasive ultrasound imaging technique, which automatically detects points of spatial mechanical inhomogeneity along the imaged artery and provides piecewise stiffness characterization. The aims of the present study are to: 1) demonstrate the initial feasibility of adaptive PWI to image the mechanical properties of an atherosclerotic plaque 2) demonstrate the feasibility to combine adaptive PWI with vector Doppler in a single imaging modality in order to simultaneously obtain information plaque mechanical properties and plaque hemodynamics. The common carotid arteries of 1 healthy subject and 2 carotid artery disease patients were scanned in vivo. One of the patients underwent carotid endarterectomy and a plaque sample was retrieved. In this patient, a higher compliance value of the stenotic segment was estimated by Adaptive PWI as compared with the adjacent arterial wall, and the healthy carotid artery. This was corroborated by histological staining of the plaque sample, which revealed the presence of a large necrotic core and a thrombus, characteristics associated with reduced stiffness. Moreover, the same sequence demonstrated the feasibility to obtain both stiffness maps and vector flow information, showing promise in atherosclerosis diagnosis and patient care.