Participation of protein metabolism in cancer progression and its potential targeting for the management of cancer.

Cancer malignancies may broadly be described as heterogeneous disorders manifested by uncontrolled cellular growth/division and proliferation. Tumor cells utilize metabolic reprogramming to accomplish the upregulated nutritional requirements for sustaining their uncontrolled growth, proliferation, and survival. Metabolic reprogramming also called altered or dysregulated metabolism undergoes modification in normal metabolic pathways for anabolic precursor's generation that serves to continue biomass formation that sustains the growth, proliferation, and survival of carcinogenic cells under a nutrition-deprived microenvironment. A wide range of dysregulated/altered metabolic pathways encompassing different metabolic regulators have been described; however, the current review is focused to explain deeply the metabolic pathways modifications inducing upregulation of proteins/amino acids metabolism. The essential modification of various metabolic cycles with their consequent outcomes meanwhile explored promising therapeutic targets playing a pivotal role in metabolic regulation and is successfully employed for effective target-specific cancer treatment. The current review is aimed to understand the metabolic reprogramming of different proteins/amino acids involved in tumor progression along with potential therapeutic perspective elucidating targeted cancer therapy via these targets.

PHF5A as a new OncoTarget and therapeutic prospects.

PHF5A (PHD-finger domain protein 5A) is a highly conserved protein comprised of 110 amino acids that belong to PHD zinc finger proteins and is ubiquitously expressed in entire eukaryotic nuclei from yeast to man. PHF5A is an essential component of the SF3B splicing complex regulating protein-protein or protein-DNA interactions; particularly involved in pre-mRNA splicing. Besides its basic spliceosome-associated attributes encompassing the regulation of alternative splicing of specific genes, PHF5A also plays a pivotal role in cell cycle regulation and morphological development of cells along with their differentiation into particular tissues/organs, DNA damage repair, maintenance of pluripotent embryonic stem cells (CSCs) embryogenesis and regulation of chromatin-mediated transcription. Presently identification of spliceosome and non-spliceosome-associated attributes of PHF5A needs great attention based on its key involvement in the pathogenesis of cancer malignancies including the prognosis of lung adenocarcinoma, endometrial adenocarcinoma, breast, and colorectal cancer. PHF5A is an essential splicing factor or cofactor actively participating as an oncogenic protein in tumorigenesis via activation of downstream signaling pathway attributed to its regulation of dysregulated splicing or abnormal alternative splicing of targeted genes. Further, the participation of PHF5A in regulating the growth of cancer stem cells might not be ignored. The current review briefly overviews the structural and functional attributes of PHF5A along with its hitherto described role in the propagation of cancer malignancies and its future concern as a potential therapeutic target for cancer management/treatment.

Precision Targeted Ablation of Fine Neurovascular Structures In Vivo Using Dual-mode Ultrasound Arrays.

Carotid bodies (CBs) are chemoreceptors that monitor and register changes in the blood, including the levels of oxygen, carbon dioxide, and pH, and regulate breathing. Enhanced activity of CBs was shown to correlate with a significant elevation in the blood pressure of patients with hypertension. CB removal or denervation were previously shown to reduce hypertension. Here we demonstrate the feasibility of a dual-mode ultrasound array (DMUA) system to safely ablate the CB in vivo in a spontaneously hypertensive rat (SHR) model of hypertension. DMUA imaging was used for guiding and monitoring focused ultrasound (FUS) energy delivered to the target region. In particular, 3D imaging was used to identify the carotid bifurcation for targeting the CBs. Intermittent, high frame rate imaging during image-guided FUS (IgFUS) delivery was used for monitoring the lesion formation. DMUA imaging provided feedback for closed-loop control (CLC) of the lesion formation process to avoid overexposure. The procedure was tolerated well in over 100 SHR and normotensive rats that received unilateral and bilateral treatments. The measured mean arterial pressure (MAP) exhibited measurable deviation from baseline 2-4 weeks post IgFUS treatment. The results suggest that the direct unilateral FUS treatment of the CB might be sufficient to reduce the blood pressure in hypertensive rats and justify further investigation in large animals and eventually in human patients.

Nonlinear Imaging of Microbubble Contrast Agent Using the Volterra Filter: In Vivo Results.

A nonlinear filtering approach to imaging the dynamics of microbubble ultrasound contrast agents (UCAs) in microvessels is presented. The approach is based on the adaptive third-order Volterra filter (TVF), which separates the linear, quadratic, and cubic components from beamformed pulse-echo ultrasound data. The TVF captures polynomial nonlinearities utilizing the full spectral components of the echo data and not from prespecified bands, e.g., second or third harmonics. This allows for imaging using broadband pulse transmission to preserve the axial resolution and the SNR. In this paper, we present the results from imaging the UCA activity in a 200- [Formula: see text] cellulose tube embedded in a tissue-mimicking phantom using a linear array diagnostic probe. The contrast enhancement was quantified by computing the contrast-to-tissue ratio (CTR) for the different imaging components, i.e., B-mode, pulse inversion (PI), and the TVF components. The temporal mean and standard deviation of the CTR values were computed for all frames in a given data set. Quadratic and cubic images, referred to as QB-mode and CB-mode, produced higher mean CTR values than B-mode, which showed improved sensitivity. Compared with PI, they produced similar or higher mean CTR values with greater spatial specificity. We also report in vivo results from imaging UCA activity in an implanted LNCaP tumor with heterogeneous perfusion. The temporal means and standard deviations of the echogenicity were evaluated in small regions with different perfusion levels in the presence and absence of UCA. The in vivo measurements behaved consistently with the corresponding calculations obtained under microflow conditions in vitro. Specifically, the nonlinear VF components produced larger increases in the temporal mean and standard deviation values compared with B-mode in regions with low to relatively high perfusion. These results showed that polynomial filters such as the TVF can provide an important tool for imaging UCA activity in regions with heterogeneous perfusion as is the case in some tumors and ischemic tissues.

In Vivo application and localization of transcranial focused ultrasound using dual-mode ultrasound arrays.

Focused ultrasound (FUS) has been proposed for a variety of transcranial applications, including neuromodulation, tumor ablation, and blood-brain barrier opening. A flurry of activity in recent years has generated encouraging results demonstrating its feasibility in these and other applications. To date, monitoring of FUS beams has been primarily accomplished using MR guidance, where both MR thermography and elastography have been used. The recent introduction of real-time dual-mode ultrasound array (DMUA) systems offers a new paradigm in transcranial focusing. In this paper, we present first experimental results of ultrasound-guided transcranial FUS (tFUS) application in a rodent brain, both ex vivo and in vivo. DMUA imaging is used for visualization of the treatment region for placement of the focal spot within the brain. This includes the detection and localization of pulsating blood vessels at or near the target point(s). In addition, DMUA imaging is used to monitor and localize the FUS-tissue interactions in real time. In particular, a concave (40 mm radius of curvature), 32-element, 3.5-MHz DMUA prototype was used for imaging and tFUS application in ex vivo and in vivo rat models. The ex vivo experiments were used to evaluate the point spread function of the transcranial DMUA imaging at various points within the brain. In addition, DMUA-based transcranial ultrasound thermography measurements were compared with thermocouple measurements of subtherapeutic tFUS heating in rat brain ex vivo. The ex vivo setting was also used to demonstrate the capability of DMUA to produce localized thermal lesions. The in vivo experiments were designed to demonstrate the ability of the DMUA to apply, monitor, and localize subtherapeutic tFUS patterns that could be beneficial in transient blood-brain barrier opening. The results show that although the DMUA focus is degraded due to the propagation through the skull, it still produces localized heating effects within a sub-millimeter volume. In addition, DMUA transcranial echo data from brain tissue allow for reliable estimation of temperature change.

Theoretical study of microbubble dynamics in sonoporation.

Sonoporation is a promising technology for promoting the transfer of drug or gene into cells using ultrasound-mediated microbubbles that transiently break up the cell membrane. In this article, a model is established to analyze the dynamics of ultrasound-mediated microbubble near the cell membrane, which may be especially useful for understanding the mechanisms of sonoporation. In the model, the velocity potential of fluid on the microbubble surface and on the cell membrane is obtained by the unsteady Bernoulli equations, and it is solved by using the boundary integral equations. By numerically analyzing the model, the typical microbubble dynamics near the cell membrane are enumerated, which may be mainly governed by mechanical index. The model also established the connections among the parameters of ultrasound exposure, microbubble characteristics, and cell membrane properties in sonoporation.

Clinical significance of upregulation of mir-196a-5p in gastric cancer and enriched KEGG pathway analysis of target genes.

BACKGROUND: miRNAs are relatively recently discovered cancer biomarkers which have important implications for cancer early diagnosis, treatment and estimation of prognosis. Here we focussed on expression of mir-196a-5p in gastric cancer tissues and cell lines so as to analyse its significance for clinicopathologic characteristics and generate enriched KEGG pathways clustered by target genes for exploring its potential roles as a biomarker in gastric cancer. MATERIALS AND METHODS: The expression of mir-196a-5p in poorly, moderate and well differentiated gastric cancer cell lines compared with GES-1 was detected by RT-qPCR, and the expression of mir-196a-5p in gastric cancer tissues comparing with adjacent non cancer tissues of 58 cases were also assessed by RT- qPCR. Subsequently, an analysis of clinical significance of mir-196a-5p in gastric cancer and enriched KEGG pathways was executed based on the miRWalk prediction database combined with bioinformatics tools DAVID 6.7 and Mirfocus 3.0. RESULTS: RT-qPCR showed that mir-196a-5p was up-regulated in 6 poorly and moderate differentiated gastric cancer cell lines SGC-7901, MKN-45, MKN-28, MGC-803, BGC-823, HGC-27 compared with GES-1, but down-regulated in the highly differentiated gastric cancer cell line AGS. Clinical data indicated mir-196a-5p to beup-regulated in gastric cancer tissues (47/58). Overexpression of mir-196a-5p was associated with more extensive degree of lymph node metastasis and clinical stage (P <0.05; x2 test). Enriched KEGG pathway analyses of predicted and validated targets in miRWalk combined with DAVID 6.7 and Mirfocus 3.0 showed that the targeted genes regulated by mir-196a-5p were involved in malignancy associated biology. CONCLUSIONS: Overexpression of mir-196a-5p is associated with lymph node metastasis and clinical stage, and enriched KEGG pathway analyses showed that targeted genes regulated by mir-196a-5p may contribute to tumorgenesis, suggesting roles as an oncogenic miRNA biomarker in gastric cancer.

Feasibility of targeting atherosclerotic plaques by high-intensity-focused ultrasound: an in vivo study.

PURPOSE: To investigate the feasibility and acute safety of targeting atherosclerotic plaques by high-intensity-focused ultrasound (US) in vivo through a noninvasive extracorporeal approach. MATERIALS AND METHODS: Four swine were included in this prospective study, three of which were familial hypercholesterolemic swine. The procedure was done under general anesthesia. After US identification of atherosclerotic plaques within the femoral arteries, plaques were targeted by high-intensity focused US with an integrated dual-mode US array system. Different ablation protocols were used to meet the study objectives, and animals were then euthanized at different time points. Targeted arterial segments were stained by hematoxylin and eosin for histopathologic examination. Numeric values are presented as means ± standard deviation. RESULTS: All swine tolerated the procedure well, with no arterial dissection, perforation, or rupture. Discrete lesions were detected in the first two swine, measuring 0.54 mm ± 0.10 and 0.25 mm ± 0.03 in cross-sectional dimensions in the first and 0.50 mm ± 0.12 and 0.24 mm ± 0.15 in the second. Confluent ablation zones were identified in the last two swine, measuring 6.92 mm and 0.93 mm in the third and 2.97 mm and 2.52 mm in the fourth. Lesions showed necrotic cores and peripheral reactive inflammatory infiltration. The endothelium overlying targeted arterial segments remained intact. CONCLUSIONS: The results demonstrate the feasibility and acute safety of targeting atherosclerotic plaques by high-intensity-focused US in vivo. Further long-term studies are needed to assess how induction of these lesions can modify the progression of atherosclerotic plaques.

Real-time implementation of a dual-mode ultrasound array system: in vivo results.

A real-time dual-mode ultrasound array (DMUA) system for imaging and therapy is described. The system utilizes a concave (40-mm radius of curvature) 3.5 MHz, 32 element array, and modular multichannel transmitter/receiver. The system is capable of operating in a variety of imaging and therapy modes (on transmit) and continuous receive on all array elements even during high-power operation. A signal chain consisting of field-programmable gate arrays and graphical processing units is used to enable real time, software-defined beamforming and image formation. Imaging data, from quality assurance phantoms as well as in vivo small- and large-animal models, are presented and discussed. Corresponding images obtained using a temporally-synchronized and spatially-aligned diagnostic probe confirm the DMUA's ability to form anatomically-correct images with sufficient contrast in an extended field of view around its geometric center. In addition, high-frame rate DMUA data also demonstrate the feasibility of detection and localization of echo changes indicative of cavitation and/or tissue boiling during high-intensity focused ultrasound exposures with 45-50 dB dynamic range. The results also show that the axial and lateral resolution of the DMUA are consistent with its f(number) and bandwidth with well-behaved speckle cell characteristics. These results point the way to a theranostic DMUA system capable of quantitative imaging of tissue property changes with high specificity to lesion formation using focused ultrasound.

Realtime control of multiple-focus phased array heating patterns based on noninvasive ultrasound thermography.

A system for the realtime generation and control of multiple-focus ultrasound phased-array heating patterns is presented. The system employs a 1-MHz, 64-element array and driving electronics capable of fine spatial and temporal control of the heating pattern. The driver is integrated with a realtime 2-D temperature imaging system implemented on a commercial scanner. The coordinates of the temperature control points are defined on B-mode guidance images from the scanner, together with the temperature set points and controller parameters. The temperature at each point is controlled by an independent proportional, integral, and derivative controller that determines the focal intensity at that point. Optimal multiple-focus synthesis is applied to generate the desired heating pattern at the control points. The controller dynamically reallocates the power available among the foci from the shared power supply upon reaching the desired temperature at each control point. Furthermore, anti-windup compensation is implemented at each control point to improve the system dynamics. In vitro experiments in tissue-mimicking phantom demonstrate the robustness of the controllers for short (2-5 s) and longer multiple-focus high-intensity focused ultrasound exposures. Thermocouple measurements in the vicinity of the control points confirm the dynamics of the temperature variations obtained through noninvasive feedback.

Real-time 2-D temperature imaging using ultrasound.

We have previously introduced methods for noninvasive estimation of temperature change using diagnostic ultrasound. The basic principle was validated both in vitro and in vivo by several groups worldwide. Some limitations remain, however, that have prevented these methods from being adopted in monitoring and guidance of minimally invasive thermal therapies, e.g., RF ablation and high-intensity-focused ultrasound (HIFU). In this letter, we present first results from a real-time system for 2-D imaging of temperature change using pulse-echo ultrasound. The front end of the system is a commercially available scanner equipped with a research interface, which allows the control of imaging sequence and access to the RF data in real time. A high-frame-rate 2-D RF acquisition mode, M2D, is used to capture the transients of tissue motion/deformations in response to pulsed HIFU. The M2D RF data is streamlined to the back end of the system, where a 2-D temperature imaging algorithm based on speckle tracking is implemented on a graphics processing unit. The real-time images of temperature change are computed on the same spatial and temporal grid of the M2D RF data, i.e., no decimation. Verification of the algorithm was performed by monitoring localized HIFU-induced heating of a tissue-mimicking elastography phantom. These results clearly demonstrate the repeatability and sensitivity of the algorithm. Furthermore, we present in vitro results demonstrating the possible use of this algorithm for imaging changes in tissue parameters due to HIFU-induced lesions. These results clearly demonstrate the value of the real-time data streaming and processing in monitoring, and guidance of minimally invasive thermotherapy.

Real-time two-dimensional temperature imaging using ultrasound.

We present a system for real-time 2D imaging of temperature change in tissue media using pulse-echo ultrasound. The frontend of the system is a SonixRP ultrasound scanner with a research interface giving us the capability of controlling the beam sequence and accessing radio frequency (RF) data in real-time. The beamformed RF data is streamlined to the backend of the system, where the data is processed using a two-dimensional temperature estimation algorithm running in the graphics processing unit (GPU). The estimated temperature is displayed in real-time providing feedback that can be used for real-time control of the heating source. Currently we have verified our system with elastography tissue mimicking phantom and in vitro porcine heart tissue, excellent repeatability and sensitivity were demonstrated.

Viscoelastic characterization of thin tissues using acoustic radiation force and model-based inversion.

By means of the viscoelastodynamic model for a two-layer solid-fluid system and a detailed account of the locally induced acoustic radiation force, a rational analytical and computational framework is established for the viscoelastic characterization of thin tissues from high-frequency ultrasound (HFUS) measurements. For practical applications, the back-analysis is set up to interpret the frequency response function, signifying the tissue's axial displacement (captured by the imaging transducer) per squared voltage driving the 'pushing' transducer, as experimental input. On parametrizing the tissue's viscoelastic behavior in terms of the standard linear model, the proposed methodology is applied to a set of measurements performed on tissue-mimicking phantom constructs with thicknesses ranging from 0.5 to 4 mm. The results demonstrate that the model-based inversion, which carefully mimics the local boundary conditions and applied ultrasound excitation, yields viscoelastic properties for the phantom that are virtually invariant over the range of specimen thicknesses tested. Beyond its immediate application to in vitro viscoelastic characterization of thin excised tissues and tissue constructs, the proposed methodology may also find use in the characterization of skin or skin lesions over bone in vivo.

Viscoelastic property measurement in thin tissue constructs using ultrasound.

We present a dual-element concave ultrasound transducer system for generating and tracking of localized tissue displacements in thin tissue constructs on rigid substrates. The system is comprised of a highly focused PZT-4 5-MHz acoustic radiation force (ARF) transducer and a confocal 25-MHz polyvinylidene fluoride imaging transducer. This allows for the generation of measurable displacements in tissue samples on rigid substrates with thickness values down to 500 microm. Impulse-like and longer duration sine-modulated ARF pulses are possible with intermittent M-mode data acquisition for displacement tracking. The operations of the ARF and imaging transducers are strictly synchronized using an integrated system for arbitrary waveform generation and data capture with a shared timebase. This allows for virtually jitter-free pulse-echo data well suited for correlation-based speckle tracking. With this technique we could faithfully capture the entire dynamics of the tissue axial deformation at pulse-repetition frequency values up to 10 kHz. Spatio-temporal maps of tissue displacements in response to a variety of modulated ARF beams were produced in tissue-mimicking elastography phantoms on rigid substrates. The frequency response was measured for phantoms with different modulus and thickness values. The frequency response exhibited resonant behavior with the resonance frequency being inversely proportional to the sample thickness. This resonant behavior can be used in obtaining high-contrast imaging using magnitude and phase response to sinusoidally modulated ARF beams. Furthermore, a second order forced harmonic oscillator (FHO) model was shown to capture this resonant behavior. Based on the FHO model, we used the extended Kalman filter (EKF) for tracking the apparent modulus and viscosity of samples subjected to dc and sinusoidally modulated ARF. The results show that the stiffness (apparent modulus) term in the FHO is largely time-invariant and can be estimated robustly using the EKF. On the other hand, the damping (apparent viscosity) is time varying. These findings were confirmed by comparing the magnitude response of the FHO (with parameters obtained using the EKF) with the measured ones for different thin tissue constructs.