Early Cancer Diagnostics Research Articles

Magnetic nanotechnologies for early cancer diagnostics with liquid biopsies: a review

Magnetic nanotechnologies for early cancer diagnostics with liquid biopsies: a review

Enzyme-free electrochemical biosensor based on double signal amplification strategy for the ultra-sensitive detection of exosomal microRNAs in biological samples

Growing evidence suggests that exosomes-encapsulated miRNAs detection is of paramount significance in early diagnostics of cancer due to protection from degradation by ribonuclease. However, exosomal microRNAs with low abundance and subtle variation have restricted their clinical application. Herein, an electrochemical biosensor for highly sensitive detection of exosomal microRNAs has been fabricated based on the double signal amplification strategy. Our proposed amplifier consists of two steps: target miRNA cyclic signal amplification induced by strand displacement reaction (SDR) and subsequent deposition of silver nanoparticles induced by streptavidin-biotin interaction. Consequently, this method shows ultrahigh sensitivity to detect miRNA. Taking miRNA-21 in exosomes as a model analyte, a detection limit of 0.4 fM (S/N = 3) can be obtained. Meanwhile, the method is relatively simple and low-cost without the requirement of enzyme, which has been applied in biological samples successfully. Therefore, our miRNA assay method has shown great promise as molecular tool in the detection of exosomal miRNA and could be widely used in clinic in the future.

Organization Decisions in Clinical Oncology Based on the Statistics of Malignant Neoplasms

Cancer service parameters in St.-Petersburg, Russia, were studied in relation to patients with urological malignant neoplasms (uMN) in 2014-2018. Material and methods. We studied a number of medical and statistical indicators obtained from the state reporting forms (ff. 7 and 35). Results. The exponential growth of absolute and “crude” indicators of morbidity and morphological verification of the MN of prostate, kidney and bladder in the early stages of the tumor is presented. An increase in the active detection of MN of the prostate gland (by 3.36) and kidney (by 2.26) were observed, due to the increase in oncological alertness at general practitioners. The number of bladder cancer patients who died within a year increased by 37.5%, indirectly indicates potentially aggressive forms of the tumor and the problem of early diagnostics of cancer. The maximum age of patients with newly diagnosed MN of the prostate/urinary bladder was 64-70 years, according to world statistics, kidney cancer diagnosed in patients of any age. Testicular MN was most often founded in patients 25-39 years old with an exponent in 2014-2016, penile MN – was verified at the age of 55-69 years, with a stable rate of increase in incidence. However, defects in the collection of primary medical information were identified, which made it difficult to evaluate cancer indicators. Conclusion. Original directions for the development of information collection medical systems and the provision of specialized medical care to the population of St-Petersburg are proposed - improving the system of registration of tumors and the quality of medical training and studies for stuff of statistical department, conducting research for prevention of the development of malignancies.

A novel xenonucleic acid mediated molecular clamping technology for early colorectal cancer diagnostics.

e16106 Background: Colorectal cancer (CRC) is one of the leading causes of cancer-related death. Early detection is critical to reduce CRC morbidity and mortality. In order to meet this need, we developed a molecular clamping assay called ColoScape for early colorectal cancer diagnostics. Methods: Nineteen mutations in four genes APC, KRAS, BRAF and CTNNB1 associated with early events in CRC pathogenesis are targeted in the ColoScape assay. Xenonucleic Acid (XNA) mediated qPCR clamping technology was applied to minimize the wild-type background amplification in order to improve assay sensitivity of CRC mutation detection. The assay analytical performance was verified and validated, cfDNA and FFPE CRC patient samples were assessed for clinical performance, and a ROC curve was applied to evaluate its performance. Results: The data showed that the assay analytical sensitivity is 0.5% Variant Allele Frequency, corresponding to ~7-8 copies of mutant DNA with 5ng total DNA input per test. This assay is highly reproducible with intra-assay CV < 3% and inter-assay CV < 5%. We have investigated 380 clinical samples including plasma cfDNA and FFPE samples from patients with different stages of CRC. The preliminary assay clinical specificity and sensitivity for cfDNA were 100% (95% CI, 80.3-97.5%) and 92.2% (95% CI, 94.7-100%) respectively with AUC being about 0.96; and 96% (95% CI, 77.6-99.7%) specificity and 92% (95% CI, 86.1-95.6%) sensitivity with AUC 0.94 for FFPE. Conclusions: XNA mediated molecular clamping assay is a rapid, precise, and sensitive assay for the detection of CRC mutations in cfDNA or FFPE samples.

Contributions from Japan to gastrointestinal endoscopy training and development: Special report.

Contributions from Japan to gastrointestinal endoscopy training and development: Special report.

New generation of oxide-based nanoparticles for the applications in early cancer detection and diagnostics

AbstractHereby the possible applications of oxide nanoparticles in the cancer diagnostics and therapy are presented. Cancer diseases are nowadays one of the most common causes of death in the highly-developed countries. Discussed will be the current clinical cancer detection methods with their shortcomings. The role of nanomedicine in cancer medicine and the potential applications of nanoparticles debated in the literature will be critically evaluated. In the second part, the most common methods for the nanoparticle synthesis will be discussed. Finally, the system for cancer detection based on the enhanced permeation-retention of multimodal high-k oxide nanoparticles doped with lanthanides will be proposed for both for themagnetic resonance imaging (non-gadolinium contrast agents) and for fluorescence guided biopsy and surgery.

Circulating tumor DNA (ctDNA) is not a good proxy for liquid biopsies of tumor tissues for early detection.

The important conclusion that ctDNA is a mediocre proxy for liquid biopsies of tumor tissues for early detection was reached after new data were published recently in Nature Genetics. These data have shown that most mutations found in ctDNA are not related to tumor tissues but rather to the precancerous condition clonal hematopoiesis. Previously, our group has analyzed the sensitivity of the ctDNA test for early detection of cancer and concluded that the achievable sensitivity, especially for small tumors, is not enough to have clinical value. Now, the new data have shown a serious compromise in specificity. We believe that scientists who are interested in early cancer diagnostics should be aware of the limitations of this test, in both sensitivity and specificity. Our work may prompt further work aiming to alleviate these important issues in the cancer diagnostics field.

Design study for a compact laser-driven source for medical x-ray fluorescence imaging

Thomson scattering sources with their hard x-ray pencil beams represent a promising candidate to drive high-resolution X-ray Fluorescence Imaging (XFI). As XFI is a scanning imaging modality, it specifically requires pencil-beam geometries along with a high beam mobility. In combination with laser-wakefield acceleration (LWFA) such sources could provide the compactness needed for a future transition into clinical application. A sufficient flux within a small bandwidth could enable in-vivo high-sensitivity XFI for early cancer diagnostics and pharmacokinetic imaging. We thus report on a specific all-laser driven source design directed at increasing the photon number within the bandwidth and opening angle defined by XFI conditions. Typical parameters of driver lasers and electron bunches from LWFA are utilized and controlled within realistic parameter regions on the basis of appropriate beam optics. An active plasma lens is implemented for chromatic focal control of the bunch. Source performance limits are identified and compared to existing x-ray sources with regard to their potential to be implemented in future clinical XFI.

Clinical application of FISH method in the early diagnostics of superficial bladder cancer

Clinical application of FISH method in the early diagnostics of superficial bladder cancer

Specialized care provided to patients with rectal cancer in Kyiv city.

The major trends in the rectal cancer incidence, detectability, morbidity, and mortality in Kyiv city as well as the state of medical care organization for cancer patients based on the statistical data of Kyiv City Clinical Oncology Center were analyzed. Despite the decrease in morbidity (‒6.3%) and mortality (‒8.1%), the cohort of rectal cancer patients in Kyiv city is increasing (+18.2%). At the same time, one-year mortality (27.0% in 2017), the detection rate at preventive check-ups, the detection rates at the early (I-II) stages are decreasing and at late-stage (IV) are rising (from 14.9% in 2011to 17.6% in 2017). The above indicates the need for improving the organization of medical care of cancer patients and early diagnostics of rectal cancer.

Regulation of multiple energy transfer processes in a simple nano-system for sensitive telomerase detection

Telomerase, as a potential biomarker for early cancer diagnostics and therapies, has attracted considerable interests concerning its detection and monitoring. Herein, we develop a novel method for sensitive detection of telomerase activity by designing a gold nanoparticles/graphene oxide (AuNPs/GO) probe. The AuNPs were functionalized with a telomerase substrate (TS) primer and a 6-carboxy-fluorescein (FAM)-modified complementary DNA (P1). In the absence of telomerase, P1 exists in the random-coiled conformation, and the fluorescence resonance energy transfer (FRET) from FAM to AuNPs and GO results in efficient fluorescence quenching. In the presence of telomerase, the multiple hybridization between TS extension products and P1 leads to the conformation transition of P1 from single-stranded DNA to double-stranded rigid structure, and thus the FRET process can be prevented with the efficient fluorescence recovery. The metal enhanced fluorescence (MEF) effect between FAM and AuNPs can further effectively enhance the fluorescence of FAM, and thus the sensitivity and specificity of telomerase detection can be remarkably improved. It is worth mentioning that the proposed strategy does not need to design complex hairpin structure and allows the measurement of telomerase activity in three crude cell extracts equivalent to 7 HeLa cells, 8 A549 cells and 8 L929 cells in 1 h. In addition, the present sensing platform can be applied to inhibitor screening, in situ telomerase imaging, and intracellular drug delivery.

Tissue ACE phenotyping in prostate cancer.

Epithelial cells of prostate express significant level of ACE and, as a result, seminal fluid has 50-fold more ACE than plasma. The substitution of highly specialized prostate epithelial cells by tumor cells results in dramatic decrease in ACE production in prostate tissues. We performed detailed characterization of ACE status in prostate tissues from patients with benign prostate hyperplasia (BPH) and prostate cancer (PC) using new approach- ACE phenotyping, that includes evaluation of: 1) ACE activity with two substrates (HHL and ZPHL); 2) the ratio of the rates of their hydrolysis (ZPHL/HHL ratio); 3) the ratio of immunoreactive ACE protein to ACE activity; 4) the pattern of mAbs binding to different epitopes on ACE – ACE conformational fingerprint - to reveal conformational changes in prostate ACE due to prostate pathology. ACE activity dramatically decreased and the ratio of immunoreactive ACE protein to ACE activity increased in PC tissues. The catalytic parameter, ZPHL/HHL ratio, increased in prostate tissues from all patients with PC, but was did not change for most |BPH patients. Nevertheless, prostate tissues of several patients diagnosed with BPH based on histology, also demonstrated decreased ACE activity and increased immunoreactive ACE protein/ACE activity and ZPHL/HHL ratios, that could be considered as more early indicators of prostate cancer development than routine histology. Thus, ACE phenotyping of prostate biopsies has a potential to be an effective approach for early diagnostics of prostate cancer or at least for differential diagnostics of BPH and PC.

Gold-nanoparticle-decorated boron-doped graphene/BDD electrode for tumor marker sensor

A boron-doped graphene (BG)/boron-doped diamond (BDD) composite film was obtained by continuously growing BDD and BG by chemical vapor deposition. In this process, the sp3 CC bonds in the diamond phase structure are directly converted into sp2 CC bonds in graphene to combine the two carbon materials, while BG nanosheets are vertically grown on the exposed planes of BDD crystals. These process features impart the vertical BG (VBG)/BDD film a large specific surface area and high electrocatalytic activity, resulting in an electroactive surface area of 2.35 cm2. Electrodeposition can be used to load a large number of Au nanoparticles (NPs) on the VBG/BDD electrode. The Au-VBG/BDD electrode was used to construct an immunosensor capable of simultaneously quantifying carcinoembryonic antigen (CEA) and carcinoma antigen 125 (CA 125) at concentrations of 0.5–100 pg mL–1 and 0.5–100 mU mL–1, respectively, with corresponding detection limits of 0.15 pg mL–1 and 0.09 mU mL–1. This method is suitable for the practical detection of CEA, CA125, and other tumor markers in human body fluids, allowing early cancer diagnostics and timely treatment.

Elucidation of Differential Nano-Textural Attributes for Normal Oral Mucosa and Pre-Cancer.

Computational analysis on altered micro-nano-textural attributes of the oral mucosa may provide precise diagnostic information about oral potentially malignant disorders (OPMDs) instead of an existing handful of qualitative reports. This study evaluated micro-nano-textural features of oral epithelium from scanning electron microscopic (SEM) images and the sub-epithelial connective tissue from light microscopic (LM) and atomic force microscopic (AFM) images for normal and OPMD (namely oral sub-mucous fibrosis, i.e., OSF). Objective textural descriptors, namely discrete wavelet transform, gray-level co-occurrence matrix (GLCM), and local binary pattern (LBP), were extracted and fed to standard classifiers. Best classification accuracy of 87.28 and 93.21%; sensitivity of 93 and 96%; specificity of 80 and 91% were achieved, respectively, for SEM and AFM. In the study groups, SEM analysis showed a significant (p < 0.01) variation for all the considered textural descriptors, while for AFM, a remarkable alteration (p < 0.01) was only found in GLCM and LBP. Interestingly, sub-epithelial collagen nanoscale and microscale textural information from AFM and LM images, respectively, were complementary, namely microlevel contrast was more in normal (0.251) than OSF (0.193), while nanolevel contrast was more in OSF (0.283) than normal (0.204). This work, thus, illustrated differential micro-nano-textural attributes for oral epithelium and sub-epithelium to distinguish OPMD precisely and may be contributory in early cancer diagnostics.

A sandwich-type electrochemical immunosensor using polythionine/AuNPs nanocomposites as label for ultrasensitive detection of carcinoembryonic antigen

A sandwich-type electrochemical immunosensor was developed in this study for sensitive detection of carcinoembryonic antigen (CEA) based on rGO/CS/AuNPs as a sensing platform and PTh-Au composites as signal labels. Reduced graphene oxide (rGO), Chitosan (CS) and AuNPs were layer by layer modified on glass carbon electrode (GCE), which effectively enhanced electron transfer and provided excellent large electrochemical active surface area. Polythionine-Au composites prepared by chemical reduction method greatly amplified electrochemical signal with high conductivity and large surface area. The prepared immunosensor showed high sensitivity, good stability and selectivity for detection of CEA with a wide range of 0.3 ng/mL to 30 ng/mL (R2 = 0.9972) and a detection limit as low as 0.1471 pg/mL. Furthermore, this sandwich-type immunosensor might be as a versatile platform for reliable early cancer diagnostics clinical.

Label-free microfluidic paper-based electrochemical aptasensor for ultrasensitive and simultaneous multiplexed detection of cancer biomarkers

Simultaneous detection of multiple tumor biomarkers in body fluids could facilitate early diagnosis of lung cancer, so as to provide scientific reference for clinical treatment. This paper depicted a multi-parameter paper-based electrochemical aptasensor for simultaneous detection of carcinoembryonic antigen (CEA) and neuron-specific enolase (NSE) in a clinical sample with high sensitivity and specificity. The paper-based device was fabricated through wax printing and screen-printing, which enabled functions of sample filtration and sample auto injection. Amino functional graphene (NG)-Thionin (THI)- gold nanoparticles (AuNPs) and Prussian blue (PB)- poly (3,4- ethylenedioxythiophene) (PEDOT)- AuNPs nanocomposites were synthesized respectively. They were used to modify the working electrodes not only for promoting the electron transfer rate, but also for immobilization of the CEA and NSE aptamers. A label-free electrochemical method was adopted, enabling a rapid simple point-of-care testing. Experimental results showed that the proposed multi-parameter aptasensor exhibited good linearity in ranges of 0.01–500 ng mL−1 for CEA (R2 = 0.989) and 0.05–500 ng mL−1 for NSE (R2 = 0.944), respectively. The limit of detection (LOD) was 2 pg mL−1 for CEA and 10 pg mL−1 for NSE. In addition, the device was evaluated using clinical serum samples and received a good correlation with large electrochemical luminescence (ECL) equipment, which would offer a new platform for early cancer diagnostics, especially in those resource-limit areas.

Towards Nanomaterials for Cancer Theranostics: A System of DNA-Modified Magnetic Nanoparticles for Detection and Suppression of RNA Marker in Cancer Cells

Theranostics of cancer using smart biocompatible materials can enable early cancer diagnostics and treatment. Here, we report on a DNA-nanoparticle functional material, which can simultaneously report the presence of an mRNA cancer biomarker and trigger its degradation in cultured cells. The nanodevice consists of two species of magnetic beads, each of which is conjugated with different components of a multicomponent deoxyribozyme (DZ) sensor. The system is activated only under two conditions: (i) in the presence of a specific target mRNA and (ii) when a magnetic field is applied. We demonstrate that delivery of such a system is markedly enhanced by the application of a magnetic field. The system not only fluorescently detects target mRNA in cultured MCF-7 cancer cells, but also induces its downregulation. Thus, the two-component magnetic nanoparticle system has characteristics of a material that can be used for cancer theranostics.

Univariate and classification analysis reveals potential diagnostic biomarkers for early stage ovarian cancer Type 1 and Type 2

Biomarkers for early detection of ovarian tumors are urgently needed. Tumors of the ovary grow within cysts and most are benign. Surgical sampling is the only way to ensure accurate diagnosis, but often leads to morbidity and loss of female hormones. The present study explored the deep proteome in well-defined sets of ovarian tumors, FIGO stage I, Type 1 (low-grade serous, mucinous, endometrioid; n = 9), Type 2 (high-grade serous; n = 9), and benign serous (n = 9) using TMT–LC–MS/MS. Data are available via ProteomeXchange with identifier PXD010939. We evaluated new bioinformatics tools in the discovery phase. This innovative selection process involved different normalizations, a combination of univariate statistics, and logistic model tree and naive Bayes tree classifiers. We identified 142 proteins by this combined approach. One biomarker panel and nine individual proteins were verified in cyst fluid and serum: transaldolase-1, fructose-bisphosphate aldolase A (ALDOA), transketolase, ceruloplasmin, mesothelin, clusterin, tenascin-XB, laminin subunit gamma-1, and mucin-16. Six of the proteins were found significant (p < .05) in cyst fluid while ALDOA was the only protein significant in serum. The biomarker panel achieved ROC AUC 0.96 and 0.57 respectively. We conclude that classification algorithms complement traditional statistical methods by selecting combinations that may be missed by standard univariate tests. SignificanceIn the discovery phase, we performed deep proteome analyses of well-defined histology subgroups of ovarian tumor cyst fluids, highly specified for stage and type (histology and grade). We present an original approach to selecting candidate biomarkers combining several normalization strategies, univariate statistics, and machine learning algorithms. The results from validation of selected proteins strengthen our prior proteomic and genomic data suggesting that cyst fluids are better than sera in early stage ovarian cancer diagnostics.

A novel cytosensor for capture, detection and release of breast cancer cells based on metal organic framework PCN-224 and DNA tetrahedron linked dual-aptamer

A convenient and efficient strategy for detecting cancer cells at preclinical stages is one of the dominant challenges in early cancer diagnostics. In this work, a sandwich-type cytosensor was structured to analyze cancer cells based on metal organic framework PCN-224 and DNA tetrahedron linked dual-aptamer. Firstly, the tetrahedral DNA nanostructures linked dual aptamers (AS1411 and MUC1) immobilizing on the gold electrode (GE) surface as bio-recognition elements can capture the MCF-7 cancer cells and improve the density and orientation of surface nanoprobes. Then, the novel metal organic framework PCN-224 homogeneously decorated by Pt nanoparticles was modified by the G-quadruplex/hemin DNAzyme, horseradish peroxidase (HRP) and dual-aptamer. The fabricated nanoprobes were applied to catalyze the oxidation of hydroquinone (HQ) with hydrogen peroxide (H2O2) for amplifying the electrochemical signal. The results indicated that the linear response of the aptasensor ranged from 20 to 1 × 107 cells/mL and the detection limit was 6 cells/mL. Finally, we performed an electrochemical reductive desorption to break goldthiol bond and release all compounds on the electrode surface for analyzing the viability of collected cells and regenerating the cytosensor. This work can be extended to other cancer cells by using specific type aptamers and has widely applications prospect in point-of-care cancer diagnosis.

Evaluation of nonlinear dynamics of the cell structure damage as a promising method for personalized cancer diagnostics

Background:Modulation interference microscopy is one of the promising technologies for early personalized cancer diagnostics, while it gives the possibility to obtain real-time images of living functioning cells at a nanometer resolution, as well as to assess spatio-temporal changes of their intracellular structures.Aim:To identify the markers of malignant transformation of cells based on a multiple scale spatio-temporal of the data obtained by lase interferometry duringin situregistration of changes in intracellular structures.Materials and methods:The study was performed with the breast adenocarcinoma MCF-7 cell line and human kidney epithelial cells HEK 293 provided by the N.N. Blokhin National Medical Research Centre of Oncology (Moscow, Russia). Changes in fluctuation of the living cell phase thickness were measured with a laser modulation interference microscope MIM-340 (PA UOMZ, Russia). The multifractal detrended fluctuation analysis (MF-DFA) was used to assess the results obtained.Results:By comparison of the scaling exponential spectra and corresponding spectra of the singular fluctuations in the minimal and maximal diameters of the phase pictures of HEK 293 cells and MCF-7 cells, it was shown that the cancer cell dynamics has definite monofractal properties: a linear dependency of the scaling exponentsτfrom the actualqparameter and “dotted” singularity spectrum (the spectral width does not exceed 0.1). The dynamics of healthy cell demonstrated multifractal properties, such as a non-linear dependencyτ(q)and a bell shaped singularity spectrum (the spectral width over 0.1).Conclusion:We have proposed a new approach to assess the intracellular structural damage during oncogenesis. The signs of monofractality are objective criteria of malignization. The results obtained seems promising as early diagnostic markers of cancer transformation (damage) of cells and determination of tumor cell sensitivity / resistance to anti-tumor agents in an individual patient.