Study protocol: Whole genome sequencing Implementation in standard Diagnostics for Every cancer patient (WIDE).

BACKGROUND: 'Precision oncology' can ensure the best suitable treatment at the right time by tailoring treatment towards individual patient and comprehensive tumour characteristics. In current molecular pathology, diagnostic tests which are part of the standard of care (SOC) only cover a limited part of the spectrum of genomic changes, and often are performed in an iterative way. This occurs at the expense of valuable patient time, available tissue sample, and interferes with 'first time right' treatment decisions. Whole Genome Sequencing (WGS) captures a near complete view of genomic characteristics of a tumour in a single test. Moreover, WGS facilitates faster implementation of new treatment relevant biomarkers. At present, WGS mainly has been applied in study settings, but its performance in a routine diagnostic setting remains to be evaluated. The WIDE study aims to investigate the feasibility and validity of WGS-based diagnostics in clinical practice. METHODS: 1200 consecutive patients in a single comprehensive cancer centre with (suspicion of) a metastasized solid tumour will be enrolled with the intention to analyse tumour tissue with WGS, in parallel to SOC diagnostics. Primary endpoints are (1) feasibility of implementation of WGS-based diagnostics into routine clinical care and (2) clinical validation of WGS by comparing identification of treatment-relevant variants between WGS and SOC molecular diagnostics. Secondary endpoints entail (1) added clinical value in terms of additional treatment options and (2) cost-effectiveness of WGS compared to SOC diagnostics through a Health Technology Assessment (HTA) analysis. Furthermore, the (3) perceived impact of WGS-based diagnostics on clinical decision making will be evaluated through questionnaires. The number of patients included in (experimental) therapies initiated based on SOC or WGS diagnostics will be reported with at least 3 months follow-up. The clinical efficacy is beyond the scope of WIDE. Key performance indicators will be evaluated after every 200 patients enrolled, and procedures optimized accordingly, to continuously improve the diagnostic performance of WGS in a routine clinical setting. DISCUSSION: WIDE will yield the optimal conditions under which WGS can be implemented in a routine molecular diagnostics setting and establish the position of WGS compared to SOC diagnostics in routine clinical care.

The effects of thermal and mechanical material properties on tumorous tissue during hyperthermia treatment.

Although there have been numerous reports in several articles about the viscoelastic properties of biological tissues, no effort has been made to investigate the combined thermal and mechanical behavior of the viscoelastic tissue. At present, the model of thermo-viscoelasticity theory with variable thermal conductivity and rheological properties of the volume is considered to investigate bio-thermo-mechanics behavior in living tissue within the context of the Lord-Shulman theory. The model is applied to a limited thickness, cancerous layer problem. The problem was solved analytically in the transformed domain using Laplace transform as a tool. The exact solution is obtained in the context of transformation Laplace. Numerical results are given and illustrated graphically for the distributions of temperature, displacement, and stress. Some correlations are produced with the results obtained for the absence of the thermal relaxation parameter. The effects of variable thermal and volume materials properties, blood perfusion rate on the behavior of various fields are examined.

The Yin-Yang Regulation of Reactive Oxygen Species and MicroRNAs in Cancer.

Reactive oxygen species (ROS) are highly reactive oxygen-containing chemical species formed as a by-product of normal aerobic respiration and also from a number of other cellular enzymatic reactions. ROS function as key mediators of cellular signaling pathways involved in proliferation, survival, apoptosis, and immune response. However, elevated and sustained ROS production promotes tumor initiation by inducing DNA damage or mutation and activates oncogenic signaling pathways to promote cancer progression. Recent studies have shown that ROS can facilitate carcinogenesis by controlling microRNA (miRNA) expression through regulating miRNA biogenesis, transcription, and epigenetic modifications. Likewise, miRNAs have been shown to control cellular ROS homeostasis by regulating the expression of proteins involved in ROS production and elimination. In this review, we summarized the significance of ROS in cancer initiation, progression, and the regulatory crosstalk between ROS and miRNAs in cancer.

Erythrocyte-cancer hybrid membrane-camouflaged melanin nanoparticles for enhancing photothermal therapy efficacy in tumors.

Cell membrane coating has emerged as an intriguing biomimetic strategy to endow nanomaterials with functions and properties inherent to source cells for various biomedical applications. Hybrid membrane of different types of cells could be coated onto nanoparticle surface to achieve additional functions. Herein, we fused red blood cell (RBC) membrane together with MCF-7 cell membrane and fabricated an erythrocyte-cancer (RBC-M) hybrid membrane-camouflaged melanin nanoparticle (Melanin@RBC-M) platform for enhancing therapeutic efficacy of photothermal therapy (PTT). The fused RBC-M hybrid membrane vesicles retained both RBC and MCF-7 cell membrane proteins and the resultant Melanin@RBC-M exhibited prolonged blood circulation and homotypic targeting to source MCF-7 cells simultaneously. Interestingly, increasing MCF-7 membrane components in RBC-M significantly enhanced the homotypic targeting function of Melanin@RBC-M while increasing RBC membrane components in RBC-M effectively reduced the cellular uptake of Melanin@RBC-M by macrophages and improved their circulation time in the blood. After intravenous injection into MCF-7 tumor-bearing athymic nude mice, Melanin@RBC-M with 1:1 membrane protein weight ratio of RBC to MCF-7 exhibited significantly higher tumor accumulation and better PTT efficacy compared with other Melanin@RBC-M with different membrane protein weight ratios as well as pristine melanin nanoparticles, due to the optimal balance between prolonged blood circulation and homotypic targeting. In addition, in vitro photoacoustic results revealed that Melanin@RBC-M had a photoacoustic signal enhancement with the increase of nanoparticle size (64 → 148 nm) and the photoacoustic amplitudes increased linearly with nanoparticle concentration at the excitation wavelength ranged from 680 nm to 800 nm, which could be used for quantification of Melanin@RBC-M in vivo. Looking forward, coating hybrid membrane onto nanoparticles could add flexibility and controllability in enhancing nanoparticles functionality and offer new opportunities for biomedical applications.

Minimizing Membrane Arachidonic Acid Content as a Strategy for Controlling Cancer: A Review.

Many cancers and pre-cancerous lesions convert membrane-bound arachidonic acid (AA) to eicosanoids that promote the survival, growth, and spread of cancer. In contrast, the long-chain omega-3s eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) can competitively inhibit AA's interaction with the enzymes that give rise to eicosanoids, while acting as precursors for alternative eicosanoids which oppose cancer development and growth. Hence, minimizing the AA content of cancer membranes, while boosting that of EPA and DHA, is a rational strategy for cancer prevention and control. The former goal can be achieved by eating a plant-based diet (inherently free of AA); by avoiding foods high in linoleic acid; by down-regulating the expression of delta-6-desaturase (D6D), rate-limiting for the conversion of linoleic acid to AA; and by competitively decreasing flux of linoleic acid through D6D with a high intake of alpha-linolenic acid (ALA) from flaxseed. ALA and DHA, potent agonists for the farnesoid X receptor, can be expected to suppress D6D transcription, and AMP-activated kinase (AMPK) activators and a cholesterol-free diet also have potential in this regard. Hence, a plant-based diet low in linoleic acid, complemented by an ample intake of flaxseed and supplemental fish oil, with or without metformin and other D6D-antagonist agents, may aid prevention and control of some cancers.

Topotecan effect on the structure of normal and cancer plasma membrane lipid models: A multi-model approach.

Topotecan is a relatively large, planar, asymmetric and polar molecule with a lactone moiety. In neutral or basic aqueous solutions, this ring opens forming the carboxylate form of Topotecan that is biologically inactive and uncapable of passively cross membranes. Nevertheless, despite this inability to cross membranes at this form, Topotecan may still be able to interact with phospholipid bilayers, disturbing them. In this context, phospholipid models, mimicking normal (DMPC at pH 7.4) and cancer cell lipid membranes (DMPC:DMPS (5:1) at pH 6.5), were used to assess structural modifications upon interaction with Topotecan. Langmuir isotherms of monolayers coupled with Brewster angle microscopy, differential scanning calorimetry of liposomes and X-ray scattering of small and wide angle of stacked multilayers were used as complementary techniques. The overall results show that the interaction of Topotecan with lipid membranes is deeply conditioned by their composition and that Topotecan seems to have a preferential interaction with the glycerol backbone of phosphatidylcholine phospholipids.

Effect of tumor properties on energy absorption, temperature mapping, and thermal dose in 13.56-MHz radiofrequency hyperthermia.

Computational techniques can enhance personalized hyperthermia-treatment planning by calculating tissue energy absorption and temperature distribution. This study determined the effect of tumor properties on energy absorption, temperature mapping, and thermal dose distribution in mild radiofrequency hyperthermia using a mouse xenograft model. We used a capacitive-heating radiofrequency hyperthermia system with an operating frequency of 13.56 MHz for in vivo mouse experiments and performed simulations on a computed tomography mouse model. Additionally, we measured the dielectric properties of the tumors and considered temperature dependence for thermal properties, metabolic heat generation, and perfusion. Our results showed that dielectric property variations were more dominant than thermal properties and other parameters, and that the measured dielectric properties provided improved temperature-mapping results relative to the property values taken from previous study. Furthermore, consideration of temperature dependency in the bio heat-transfer model allowed elucidation of precise thermal-dose calculations. These results suggested that this method might contribute to effective thermoradiotherapy planning in clinics.

Targeting of herbal bioactives through folate receptors: a novel concept to enhance intracellular drug delivery in cancer therapy.

Targeted drug delivery through folate receptor (FR) has emerged as a most biocompatible, target oriented, and non-immunogenic cargoes for the delivery of anticancer drugs. FRs are highly overexpressed in many tumor cells (like ovarian, lung, breast, kidney, brain, endometrial, and colon cancer), and targeting them through conjugates bearing specific ligand with encapsulated nanodrug moiety is undoubtedly, a promising approach toward tumor targeting. Folate, being an endogenous ligand, can be exploited well to affect various cellular events occurring during the progress of tumor, in a more natural and definite way. Thus, the aim of the review lies in summarizing the advancements taken place in the drug delivery system of different therapeutics through FRs and to refine its role as an endogenous ligand, in targeting of synthetic as well as natural bioactives. The review also provides an update on the patents received on the folate-based drug delivery system.

Assessing the Benefits of Drug Delivery by Nanocarriers: A Partico/Pharmacokinetic Framework.

OBJECTIVE: An in vivo kinetic framework is introduced to analyze and predict the quantitative advantage of using nanocarriers to deliver drugs, especially anticancer agents, compared to administering the same drugs in their free form. METHODS: This framework recognizes three levels of kinetics. First is the particokinetics associated with deposition of nanocarriers into tissues associated with drug effect and toxicity, their residence inside those tissues, and elimination of the nanocarriers from the body. Second is the release pattern in time of free drug from the nanocarriers. Third is the pharmacokinetics of free drug, as it relates to deposition and elimination processes in the target and toxicity associated tissues, and total body clearance. A figure of merit, the drug targeting index (DTI), is used to quantitate the benefit of nanocarrier-based drug delivery by considering the effects of preferential deposition of nanoparticles into target tissues and relative avoidance of tissues associated with drug toxicity, compared to drug that is administered in its free form. RESULTS: General methods are derived for calculating DTI when appropriate particokinetic, pharmacokinetic, and drug release rate information is available, and it is shown that relatively simple algebraic forms result when some common assumptions are made. CONCLUSION: This approach may find use in developing and selecting nanocarrier formulations, either for populations or for individuals.

New non-invasive safe, quick, economical method of detecting various cancers was found using QRS complex or rising part of T-wave of recorded ECGs. Cancers can be screened along with their biochemical parameters & therapeutic effects of any cancer treatments can be evaluated using recorded ECGs of the same individual.

There are many methods of detecting cancers including detection of cancer markers by blood test, (which is invasive, time consuming and relatively expensive), detection of cancers by non-invasive methods such as X-Ray, CT scan, and MRI & PET Scan (which are non-invasive and quick but very expensive). Our research was performed to develop new non-invasive, safe, quick economical method of detecting cancers and the 1st author already developed for clinically important non-invasive new methods including early stage of present method using his method of localizing accurate organ representation areas of face, eyebrows, upper lip, lower lip, surface and dorsal part of the tongue, surface backs, and palm side of the hands. This accurate localization of the organ representation area of the different parts of the body was performed using electromagnetic field resonance phenomenon between 2 identical molecules or tissues based on our US patented non-invasive method in 1993. Since year 2000, we developed the following non-invasive diagnostic methods that can be quickly identified by the patented simple non-invasive method without using expensive or bulky instrument at any office or field where there is no electricity or instrument available. The following are examples of non-invasive quick method of diagnosis and treatment of cancers using different approaches: 1) Soft red laser beam scanning of different parts of body; 2) By speaking voice; 3) Visible and invisible characteristic abnormalities on different organ representation areas of the different parts of the body, and 4) Mouth, Hand, and Foot Writings of both right and left side of the body. As a consequence of our latest research, we were able to develop a simple method of detecting cancer from existing recorded electrocardiograms. In this article, we are going to describe the method and result of clinical applications on many different cancers of different organs including lung, esophagus, breast, stomach, colon, uterus, ovary, prostate gland, as well as common bone marrow related malignancies such as Hodgkin's Lymphoma, Non-Hodgkin's Lymphoma, Multiple Myeloma as well as Leukemia.

Time-resolved ICP-MS analysis of mineral element contents and distribution patterns in single cells.

Novel single cell techniques are attracting growing interest for clinical applications, because they can elucidate the cellular diversity and heterogeneity instead of the average masked by bulk measurements. Herein, time-resolved ICP-MS for the determination of essential mineral elements in single cells has been developed and is used to analyze the contents and distribution patterns of Fe, Cu, Zn, Mn, P and S in two types of cancer cells (HeLa and A549) and one type of normal cells (16HBE). The results show that there are obvious differences in contents and distribution patterns of the elements among the three types of cells. The mass of Fe, Zn, Cu, Mn, P, and S in individual HeLa cells is significantly higher and span a broader range of values than in the single 16HBE and A549 cells. The contents of Fe, Zn, and Cu follow log-normal distributions, and Mn, P, and S follow Poisson distributions with high λ values in single HeLa cells, indicating a large cell-to-cell variance. Comparatively, the contents of Cu, Zn, P, and S in 16HBE cells show the narrowest distribution range among the three tested cells, demonstrating the homogenous distribution of the elements in the cells. The method of single cell ICP-MS (SC-ICP-MS) provides potential applications for the monitoring of the variation of mineral elements at a single cell level.

The antimitotic potential of PARP inhibitors, an unexplored therapeutic alternative.

ADP-ribosylation or PARsylation is one of the most abundant modifications of proteins and DNA. Although the usual context for PARsylation involves the detection and repair of DNA damage in the cell, poly(ADP-ribose) polymerases are known to regulate a number of biological processes besides maintaining genome integrity. One of these processes is the assembly and maintenance of the mitotic spindle where the presence of PARP-1 and tankyrase 1 (TNKS1), two of the best-characterized members of the PARP superfamily, is of critical importance. Here, we recapitulate the biological implications of the absence of poly(ADP-ribose) polymerases and depletion of PARsylation occurrence in mitosis in order to better understand the antimitotic effects of PARP inhibitors. In this regard, we also present an overview of the existing and more relevant molecules, with a special attention to the historical development of their pharmacological properties and structures, as well as a brief summary of clinical trials involving PARP inhibitors.

Target driven preclinical screening for new antimitotic chemotherapy agents.

Currently approved antimitotic therapies used in chemotherapy are microtubule-targeting agents (MTAs). Despite they achieved some level of success, they have limited efficacy as single agents, with issues of slippages and resistance, and cause significant side effects. The advances in the identification of other mitosis-related targets led to the development of new mitotic regulators aimed to perturb mitosis without interfering with microtubule dynamics in non-dividing cells trying to reduce side effects in patients. Some of these compounds like those targeted to entry and mitotic kinases, mitotic kinesins/motor proteins, and multiprotein complexes have been evaluated in vitro and in animal models, and some of them have reached clinical trials. Despite promising preclinical results, in many cases, the efficacy demonstrated by these new antimitotics was not better than current microtubule inhibitors. In this paper we review present and future strategies on the search for new antimitotic compounds based on identification of new protein targets and development of multifunctional inhibitors of mitosis in cancer cells.

Bromelain surface modification increases the diffusion of silica nanoparticles in the tumor extracellular matrix.

Tumor extracellular matrix (ECM) represents a major obstacle to the diffusion of therapeutics and drug delivery systems in cancer parenchyma. This biological barrier limits the efficacy of promising therapeutic approaches including the delivery of siRNA or agents intended for thermoablation. After extravasation due to the enhanced penetration and retention effect of tumor vasculature, typical nanotherapeutics are unable to reach the nonvascularized and anoxic regions deep within cancer parenchyma. Here, we developed a simple method to provide mesoporous silica nanoparticles (MSN) with a proteolytic surface. To this extent, we chose to conjugate MSN to Bromelain (Br-MSN), a crude enzymatic complex, purified from pineapple stems, that belongs to the peptidase papain family. This surface modification increased particle uptake in endothelial, macrophage, and cancer cell lines with minimal impact on cellular viability. Most importantly Br-MSN showed an increased ability to digest and diffuse in tumor ECM in vitro and in vivo.

6-iodolactone, key mediator of antitumoral properties of iodine.

An iodinated derivative of arachidonic acid, 5-hydroxy-6-iodo-8,11,14-eicosatrienoic acid, δ-lactone (6-IL) has been implicated as a possible intermediate in the autoregulation of the thyroid gland by iodine. In addition to antiproliferative and apoptotic effects observed in thyrocytes, this iodolipid could also exert similar actions in cells derived from extrathyroidal tissues like mammary gland, prostate, colon, or the nervous system. In mammary cancer (solid tumors or tumor cell lines), 6-IL has been detected after molecular iodine (I2) supplement, and is a potent activator of peroxisome proliferator-activated receptor type gamma (PPARγ). These observations led us to propose I2 supplement as a novel coadjutant therapy which, by inducing differentiation mechanisms, decreases tumor progression and prevents chemoresistance. Some kinds of tumoral cells, in contrast to normal cells, contain high concentrations of arachidonic acid, making the I2 supplement a potential "magic bullet" that enables local, specific production of 6-IL, which then exerts antineoplastic actions with minimal deleterious effects on normal tissues.

Mesoporous silica nanoparticles in medicine--recent advances.

MSNs have attracted increasing interest as drug carriers due to promising in vivo results in small-animal disease models, especially related to cancer therapy. In most cases small hydrophobic drugs have been used, but recent in vitro studies demonstrate that MSNs are highly interesting for gene delivery applications. This review covers recent advances related to the therapeutic use of mesoporous silica nanoparticles (MSNs) administered intravenously, intraperitoneally or locally. We also cover the use of MSNs in alternative modes of therapy such as photodynamic therapy and multidrug therapy. We further discuss the current understanding about the biodistribution and safety of MSNs. Finally, we critically discuss burning questions especially related to experimental design of in vivo studies in order to enable a fast transition to clinical trials of this promising drug delivery platform.

Galectin 3-ß-galactobiose interactions.

Force spectroscopy has been used to investigate the interaction between the disaccharide ß-galactobiose and the pro-metastatic regulatory protein galectin-3 (Gal3). The studies revealed specific interactions characterised by an off-rate dissociation constant k(off)=0.33 s(-1) and interaction distance x=0.2 nm at zero applied force. These data suggest a lifetime for the interaction of 3.0 s. The results are consistent with the hypothesis that oral consumption of modified citrus pectin controls cancer metastasis by inhibiting the role of Gal3. The modification is considered to facilitate binding of pectin-derived galactan sidechains to Gal3 and inhibition of the roles of Gal3 as a pro-metastatic regulatory protein.

[Comparative study on tumor cell apoptosis in vitro induced by extracts of Stellera chamaejasme].

OBJECTIVE: To in vitro compare the induction of extracts of Stellera chamaejasme ESC, ESC-1 and ESC-2 on NCI-H157 cell apoptotic. METHOD: The apoptosis rate was inspected by flow cytometry; caspase-3, 8, 9 activities was measured by spectrophotometry. Fas, Fas-L, TNF-alpha, Trail-R, Cyto-C, Smac/diablo protein expressions of apoptosis pathway was observed by Elisa method. RESULT: Compared with the control group, ESC, ESC-1, ESC-2 can significantly improve the apoptosis rate of NCI-H157 cell. ESC significantly improved cells caspase-3, 8 activity, ESC-2 can significantly improve the activity of caspase-3, 8, 9. ESC, ESC-1, ESC-2 significantly increased Fas expression and ESC significantly increased Fas/Fas-L ratio. ESC, ESC-1, ESC-2 significantly increased TNF-alpha protein expression. ESC-1 significantly lowered TRAIL-R expression. ESC, ESC-1, ESC-2 had no significant effect on Cyto-C. ESC-1, ESC-2 significantly reduced Smac protein expression. CONCLUSION: The apoptotic effect induced by ESCs may be related to the regulation of death receptor pathway proteins. Induction mechanisms of ESCs were so complicated that it may have a two-way regulatory effect. Its induction in apoptosis is a result from comprehensive regulation and control.

Multifunctionality of indocyanine green-loaded biodegradable nanoparticles for enhanced optical imaging and hyperthermia intervention of cancer.

The aim of this study was to develop and characterize multifunctional biodegradable and biocompatible poly lactic-co-glycolic acid (PLGA) nanoparticles loaded with indocyanine green (ICG) as an optical-imaging contrast agent for cancer imaging and as a photothermal therapy agent for cancer treatment. PLGA-ICG nanoparticles (PIN) were synthesized with a particle diameter of 246±11 nm, a polydispersity index of 0.10±0.03, and ICG loading efficiency of 48.75±5.48%. PIN were optically characterized with peak excitation and emission at 765 and 810±5 nm, a fluorescence lifetime of 0.30±0.01 ns, and peak absorbance at 780 nm. The cytocompatibility study of PIN showed 85% cell viability till 1-mg/ml concentration of PIN. Successful cellular uptake of ligand conjugated PIN by prostate cancer cells (PC3) was also obtained. Both phantom-based and in vitro cell culture results demonstrated that PIN (1) have the great potential to induce local hyperthermia (i.e., temperature increase of 8 to 10°C) in tissue within 5 mm both in radius and in depth; (2) result in improved optical stability, excellent biocompatibility with healthy cells, and a great targeting capability; (3) have the ability to serve as an image contrast agent for deep-tissue imaging in diffuse optical tomography.