Detection Of Tumor Biomarkers Research Articles

Aptamer-Based Chemiluminescent Optical Fiber Immunosensor with Enhanced Signal Amplification for Ultrasensitive Detection of Tumor Biomarkers

Aptamer-Based Chemiluminescent Optical Fiber Immunosensor with Enhanced Signal Amplification for Ultrasensitive Detection of Tumor Biomarkers

Fluorescence turn-on immunosensing of HE4 biomarker and ovarian cancer cells based on target-triggered metal-enhanced fluorescence of carbon dots

Rapid and sensitive detection of tumor biomarkers and cancer cells is of crucial importance for the early diagnosis and prognosis prediction of cancer. The present report describes a target-induced fluorescence enhancement immunosensor that utilizes the optical property of carbon dots (CDs) and the metal-enhanced fluorescence effect (MEF) property of silver nanoparticles (AgNPs) for the sensitive detection of the cancer biomarker human epididymis protein 4 (HE4) and ovarian cancer cells. Nitrogen and sulfur co-doped CDs with a quantum yield of 85.6% were prepared and served as the fluorophore in MEF. The HE4 antibody (Ab) specific to the HE4 antigen was linked covalently to the surface of the synthesized CDs as the capture. The HE4 Ab-conjugated AgNPs (AgNPs-Ab) were prepared and utilized as signal amplification elements. In the presence of the target HE4, composite sandwich structures were formed between the labeled CDs-Ab and AgNPs-Ab, which brought the CDs and AgNPs into proximity, resulting in the fluorescence of CDs enhancement owing to MEF. The intensity of fluorescence enhancement was positively correlated with the HE4 concentration in the clinically important range of 0.01–200 nM with a limit detection of 2.3 pM. Moreover, the immunosensor was also successfully applied to specific fluorescence labeling and quantitative determination of HE4-positive ovarian cancer cells. The proposed target-triggered MEF sensor platform demonstrated high sensitivity, excellent anti-interference ability, along with successful validation in complex biological matrices, providing a new approach for HE4 detection in early diagnosis and therapeutic monitoring.

Ultrasensitive electrochemical detection of circulating tumor DNA by hollow polymeric nanospheres and dual enzyme assisted target amplification strategy

Detection of tumor biomarker plays a crucial role in the early diagnosis and treatment of cancer. Herein, hollow polymeric nanospheres with a high loading efficiency of ferrocene molecules (Fc-HPNs) were prepared by self-assembly route between polyethylenimine-Fc (PEI-Fc) and poly acrylic acid (PAA) on SiO2 nanoparticles. Using Fc-HPNs as an efficient electrochemical tag and combining dual-enzyme assisted target amplification strategy, an ultrasensitive electrochemical biosensor for the detection of circulating tumor DNA (ctDNA) was developed. The electrochemical signals were greatly enhanced due to the high loading of Fc tags on the hollow polymeric nanospheres and the catalytic effect of ascorbic acid on the reduction reaction of Fc. Furthermore, the use of the magnetic electrode avoided the tedious and complex modification process of the traditional electrode, which made the fabrication of the electrochemical biosensor facile and time-saving. The developed biosensor exhibited a fine linear response to ctDNA concentrations from 10 fM to 10 nM with a low detection limit of 1.6 fM, which shows great potential in early-stage diagnosis and treatment of cancer.

Nanotechnology in Tumor Biomarker Detection: The Potential of Liganded Nanoclusters as Nonlinear Optical Contrast Agents for Molecular Diagnostics of Cancer.

Simple SummaryThe key factor in preventing premature death from cancer is an early and accurate diagnosis. While common diagnostic procedures are successful in the detection and rough description of a tumor mass, a deeper insight into cancer’s molecular features is needed to optimize the treatment and increase the chances of survival. Nanotechnology can aid the molecular diagnostics of cancers through a design of nanomaterials that can simultaneously recognize specific cancer-associated molecules, so-called tumor biomarkers, and allow for their visualization by different imaging techniques. With a recent explosion in the production of various nanomaterials, the selection of the most suitable nanomaterial for tumor biomarker detection becomes a challenge. In this article, we review recent advances in the molecular diagnostics of cancer using nanotechnology and focus on liganded noble metal quantum nanoclusters, a class of ultrasmall nanomaterials with distinctive structural and optical properties, as tools in tumor biomarker detection.Cancer is one of the leading causes of premature death, and, as such, it can be prevented by developing strategies for early and accurate diagnosis. Cancer diagnostics has evolved from the macroscopic detection of malignant tissues to the fine analysis of tumor biomarkers using personalized medicine approaches. Recently, various nanomaterials have been introduced into the molecular diagnostics of cancer. This has resulted in a number of tumor biomarkers that have been detected in vitro and in vivo using nanodevices and corresponding imaging techniques. Atomically precise ligand-protected noble metal quantum nanoclusters represent an interesting class of nanomaterials with a great potential for the detection of tumor biomarkers. They are characterized by high biocompatibility, low toxicity, and suitability for controlled functionalization with moieties specifically recognizing tumor biomarkers. Their non-linear optical properties are of particular importance as they enable the visualization of nanocluster-labeled tumor biomarkers using non-linear optical techniques such as two-photon-excited fluorescence and second harmonic generation. This article reviews liganded nanoclusters among the different nanomaterials used for molecular cancer diagnosis and the relevance of this new class of nanomaterials as non-linear optical probe and contrast agents.

PIWI-interacting RNAs are aberrantly expressed and may serve as novel biomarkers for diagnosis of lung adenocarcinoma.

BackgroundLung adenocarcinoma (LUAD) is the main subtype of primary lung cancer and is a leading cause of cancer‐related death worldwide. PIWI‐interacting RNAs (piRNAs) are a type of small non‐coding RNAs that may play crucial roles in cancer progression and serve as biomarkers for tumor detection. This study aimed to explore the expression profiles and diagnostic values of piRNAs in LUAD.MethodsSmall RNA sequencing was performed to investigate tissue piRNA profiles of LUAD. The expression of selected upregulated piRNAs were detected in tissues and serum exosome samples by quantitative real‐time polymerase chain reaction (qRT‐PCR). Serum exosomes were identified by transmission electron microscope, nanoparticle tracking analysis, and western blot analysis. Receiver operating characteristic (ROC) curve was adopted to quantify the diagnostic potentials of piRNAs in LUAD. Finally, a piRNA panel was developed by multivariate logistic regression model.ResultsWe identified that 76 piRNAs were overexpressed and 9 piRNAs were underexpressed in LUAD tissues compared with adjacent non‐tumor tissues. Among the top 10 overexpressed piRNAs, 4 piRNAs (piR‐hsa‐26925, piR‐hsa‐5444, piR‐hsa‐30636, and piR‐hsa‐8757) were verified by qRT‐PCR to be significantly upregulated in LUAD tissues. Moreover, piR‐hsa‐26925 and piR‐hsa‐5444 had a significantly higher level in serum exosome samples of LUAD patients than those of healthy controls. We finally established a 2‐piRNA panel composed of piR‐hsa‐26925 and piR‐hsa‐5444, which showed higher diagnostic performance for LUAD with an AUC of 0.833.ConclusionsOur finding revealed the abnormally expressed piRNAs in LUAD, and serum exosomal piR‐hsa‐26925 and piR‐hsa‐5444 could serve as potential biomarkers for LUAD diagnosis.

Biomimetic Nanostructure Platform for Cancer Diagnosis Based on Tumor Biomarkers.

Biomarker discovery and its clinical use have attracted considerable attention since early cancer diagnosis can significantly decrease mortality. Cancer biomarkers include a wide range of biomolecules, such as nucleic acids, proteins, metabolites, sugars, and cytogenetic substances present in human biofluids. Except for free-circulating biomarkers, tumor-extracellular vesicles (tEVs) and circulating tumor cells (CTCs) can serve as biomarkers for the diagnosis and prognosis of various cancers. Considering the potential of tumor biomarkers in clinical settings, several bioinspired detection systems based on nanotechnologies are in the spotlight for detection. However, tremendous challenges remain in detection because of massive contamination, unstable signal-to-noise ratios due to heterogeneity, nonspecific bindings, or a lack of efficient amplification. To date, many approaches are under development to improve the sensitivity and specificity of tumor biomarker isolation and detection. Particularly, the exploration of natural materials in biological frames has encouraged researchers to develop new bioinspired and biomimetic nanostructures, which can mimic the natural processes to facilitate biomarker capture and detection in clinical settings. These platforms have substantial influence in biomedical applications, owing to their capture ability, significant contrast increase, high sensitivity, and specificity. In this review, we first describe the potential of tumor biomarkers in a liquid biopsy and then provide an overview of the progress of biomimetic nanostructure platforms to isolate and detect tumor biomarkers, including in vitro and in vivo studies. Capture efficiency, scale, amplification, sensitivity, and specificity are the criteria that will be further discussed for evaluating the capability of platforms. Bioinspired and biomimetic systems appear to have a bright future to settle obstacles encountered in tumor biomarker detection, thus enhancing effective cancer diagnosis.

Liquid biopsy in cholangiocarcinoma: Current status and future perspectives.

Cholangiocarcinoma (CCA) are a heterogeneous group of tumors in terms of aetiology, natural history, morphological subtypes, molecular alterations and management, but all sharing complex diagnosis, management, and poor prognosis. Several mutated genes and epigenetic changes have been detected in CCA, with the potential to identify diagnostic and prognostic biomarkers and therapeutic targets. Accessing tumoral components and genetic material is therefore crucial for the diagnosis, management and selection of targeted therapies; but sampling tumor tissue, when possible, is often risky and difficult to be repeated at different time points. Liquid biopsy (LB) represents a way to overcome these issues and comprises a diverse group of methodologies centering around detection of tumor biomarkers from fluid samples. Compared to the traditional tissue sampling methods LB is less invasive and can be serially repeated, allowing a real-time monitoring of the tumor genetic profile or the response to therapy. In this review, we analysis the current evidence on the possible roles of LB (circulating DNA, circulating RNA, exosomes, cytokines) in the diagnosis and management of patients affected by CCA.

Abstract PO-046: MethylationToActivity: A deep-learning framework that reveals promoter activity landscapes from DNA methylomes in individual tumors

Abstract Although genome-wide DNA methylomes have demonstrated their clinical value as reliable biomarkers for tumor detection, subtyping, and classification, their direct biological impacts at the individual gene level remain elusive. Here we present MethylationToActivity (M2A), a machine learning framework that uses convolutional neural networks to infer promoter activities based on H3K4me3 and H3K27ac enrichment, from DNA methylation patterns for individual genes. Using publicly available datasets in real-world test scenarios, we demonstrate that M2A is highly accurate and robust in revealing promoter activity landscapes in various pediatric and adult cancers, including both solid and hematologic malignant neoplasms. Citation Format: Justin Williams, Beisi Xu, Daniel Putnam, Andrew Thrasher, Chunliang Li, Jun Yang, Xiang Chen. MethylationToActivity: A deep-learning framework that reveals promoter activity landscapes from DNA methylomes in individual tumors [abstract]. In: Proceedings of the AACR Virtual Special Conference on Artificial Intelligence, Diagnosis, and Imaging; 2021 Jan 13-14. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(5_Suppl):Abstract nr PO-046.

MethylationToActivity: a deep-learning framework that reveals promoter activity landscapes from DNA methylomes in individual tumors

Although genome-wide DNA methylomes have demonstrated their clinical value as reliable biomarkers for tumor detection, subtyping, and classification, their direct biological impacts at the individual gene level remain elusive. Here we present MethylationToActivity (M2A), a machine learning framework that uses convolutional neural networks to infer promoter activities based on H3K4me3 and H3K27ac enrichment, from DNA methylation patterns for individual genes. Using publicly available datasets in real-world test scenarios, we demonstrate that M2A is highly accurate and robust in revealing promoter activity landscapes in various pediatric and adult cancers, including both solid and hematologic malignant neoplasms.

Research advance of liquid biopsy in gastrointestinal stromal tumors

Gastrointestinal stromal tumor (GIST) is the most common mesenchymal tumors in the gastrointestinal tract. Surgical resection is the only curative treatment, while imatinib is the first-line therapy for recurrent, metastatic, and unresectable GIST. However, more than half of GIST patients suffer from secondary resistance to imatinib within 2 years after treatment initiation. Therefore, early diagnosis, drug resistance and recurrence surveillance are critical for GIST patients. Liquid biopsy is a new method which utilizes the detection of tumor biomarkers in peripheral blood for early diagnosis and therapeutic efficacy assessment. In recent years, liquid biopsy has achieved significant research progress in several kinds of malignancy. This review aims at presenting an overview on research advance of liquid biopsy in GIST and may provide a new method for early diagnosis and therapeutic efficacy assessment of GIST.

Intratumoral and intertumoral heterogeneity of HER2 immunohistochemical expression in gastric cancer

BackgroundGiven the high heterogeneity of tumor tissue in gastric cancer (GC), inaccurate detection of tumor biomarkers will inevitably hamper a precise diagnosis and selection of patients for targeted therapies. Human epidermal growth factor receptor 2 (HER2) has been widely accepted as an underlying treatment biomarker of GC. The objective of this study is to investigate the heterogeneity (both intratumoral and intertumoral) of HER2 expression in GC, and the relationship between heterogeneity and the clinicopathological features. MethodsA total of 618 patients with primary gastric adenocarcinoma were recruited, and two formalin-fixed paraffin-embedded (FFPE) tumor-containing blocks of each patient were selected for HER2 immunohistochemical (IHC) assay. Clinicopathological characteristics were recorded, and intratumoral and intertumoral heterogeneity of HER2 IHC expression was determined. ResultsThe results indicated that the dual-block assays significantly increased the HER2 overexpression (IHC 2+ and 3+) rate compared with the single paraffin block detection. Approximately 50 % of the cases showed intratumoral HER2 heterogeneity within a single tissue section, and 30.10 % of cases showed intertumoral heterogeneity between a patient’s two blocks. Furthermore, intertumoral heterogeneity was associated with tumors of small size (P = 0.029) and distal location (P = 0.032) characters, while the intratumoral heterogeneity was correlated with poorly differentiated carcinomas. Laurén’s diffuse type showed a notably higher intratumoral heterogeneity rate, and the mixed type exhibited higher intertumoral HER2 discordance between the dual-block cohorts (P < 0.001). Besides, HER2 heterogenous overexpression was not associated with age, gender, type of resection, lymphatic or venous invasion, perineural invasion or pTNM (P > 0.05) for both cohorts. ConclusionThe research findings in this paper indicate that the intratumoral and intertumoral heterogeneity of HER2 overexpression is common in GC patients, and these variations are associated with certain clinicopathological features. We highly recommend multi-block HER2 assessment for accurate diagnosis of GC.

Magnetic immunoassay for tumor clinical diagnosis based on rolling circular amplification (RCA) coupled with ICP-MS

Herein, we constructed magnetic immunoassay-based diagnostic platform for cancer biomarker of patient’ s sample by coupling rolling circular amplification with ICP-MS. Tumor biomarker (alpha-fetoprotein, AFP, Carcinoembryonic antigen, CEA) from clinical sample was initially captured by biotinylated antibody functionalized on the surface of magnetic beads, and then recognized by a secondary antibody that linked with a RCA(Rolling Circular Amplification)-primer. Up addition of RCA-template and DNA polymerase, one single target recognition event can be converted into multiple predesigned DNA sequences. This specific sequence can be hybridized with DNA-conjugated AuNPs and form a DNA-AuNP complex for ICP-MS measurement. Under the optimal experimental parameters, this platform can detect AFP and CEA in 20% serum, as low as 2 pg/mL and 5 pg/mL, respectively, which rivalled ELISA. We anticipate this methodology for tumor biomarker detection can be used for clinical diagnosis in future.

Multiplexed detection of tumor biomarkers utilizing immunohybridization Chain reaction integrated photonic barcodes

Multiplexed detection of cancer markers is of great value in clinical diagnosis. Herein, we present a new immunoassay method for simultaneous quantification of multiple tumor biomarkers. Our approach integrates photonic crystals (PhC) barcodes with DNA hybridization chain reaction (HCR) to amplify signal. Target biomarkers are captured by primary antibodies which are immobilized onto the PhC barcodes, forming sandwich immunocomplex with the assistance of second antibodies that are conjugated with DNA initiator. Subsequently, a chain reaction of hybridization events is propagated in attendance of a pair of complementary FAM-decorated H1 and H2 hairpin probes. As a result, the PhC barcodes exhibit multiplex encoding ability for different markers. By utilizing PhC barcodes with HCR for immunoassay, low-abundance cancer biomarkers (CEA, AFP and PSA) can be screened with high detection sensitivity and specificity. Our results showed a wide range from 0.001 to 1000 ng/mL; and the detection limit as low as 0.50 pg/mL. Our approach was also evaluated for analyzing human serum sample with considerable accuracy, which implies its potential as a new platform for simultaneous multiplex immunoassay.

On the Electrochemical Detection of Alpha-Fetoprotein Using Aptamers: DNA Isothermal Amplification Strategies to Improve the Performance of Weak Aptamers.

Affinity characterization is essential to develop reliable aptamers for tumor biomarker detection. For alpha-fetoprotein (AFP), a biomarker of hepatocellular carcinoma (HCC), two DNA aptamers were described with very different affinity. In this work, we estimate the dissociation constant of both of them by means of a direct assay on magnetic beads modified with AFP and electrochemical detection on carbon screen-printed electrodes (SPCE). Unlike previous works, both aptamers showed similar dissociation constant (Kd) values, in the subµM range. In order to improve the performance of these aptamers, we proposed the isothermal amplification of the aptamers by both terminal deoxynucleotidyl transferase (TdT) and rolling circle amplification (RCA). Both DNA amplifications improved the sensitivity and also the apparent binding constants from 713 nM to 189 nM for the short aptamer and from 526 nM to 32 nM for the long aptamer. This improvement depends on the true affinity of the binding pair, which ultimately limits the analytical usefulness.

Label-Free Amperometric Immunosensor Based on Au@Pt DNPs/MoSe2@MoS2 with Multiple Signal Amplification Capabilities for Squamous Cell Carcinoma Antigen Detection

Herein, urchin-like gold@platinum dendritic nanoparticles supported on molybdenum selenide@molybdenum disulfide (Au@Pt DNPs/MoSe2@MoS2) nanocomposite with multiple signal amplification capabilities was successfully synthesized as the matrix platform of label-free amperometric immunosensor for the quantitative detection of squamous cell carcinoma antigen (SCCA). Au@Pt DNPs/MoSe2@MoS2 consists of Au@Pt DNPs with splendid biocompatibility as well as catalysis and MoSe2@MoS2 with reformative peroxidase activity, which owned enhanced electrocatalysis toward hydrogen peroxide reduction and steady bioconjugation for bioprotein immobilization. Additionally, Au@Pt DNPs/MoSe2@MoS2 possessed satisfactory conductivity to accelerate electron transfer at the electrode interface. With the help of Au@Pt DNPs/MoSe2@MoS2, the immunosensor procured a good linear extent of SCCA detection between 100 fg ml−1 and 100 ng ml−1, reached a lower detection limit than previous reports. Moreover, the immunosensor with good stability and reproducibility achieved the specific detection of SCCA in the mixture containing interferents as well as in the real human serum sample, which testified to the proposed amperometric immunosensor was reliable in clinical detection of tumor biomarkers.

Synthesis of raspberry-like nanogapped Fe3O4@Au nanocomposites for SERS-based lateral flow detection of multiple tumor biomarkers

A novel magnetic SERS-LFA system with raspberry-like Fe3O4@Au MNPs as nanogapped SERS tags was proposed for the quantitative and ultra-sensitive detection of multiple tumor biomarkers in biological samples.

An amperometric biosensor based on Cu2O@Au nanocomposites for the detection of galectin-1 via lactose–galectin interactions

In this work, a novel label-free electrochemical biosensor is developed for the detection of galectin-1 (Gal-1) based on gold nanoparticle (AuNP) loaded octahedral Cu2O (Cu2O@Au) nanocomposites. The AuNPs on the surface of the Cu2O nanocrystals not only enhance the electrochemical performance, but also serve as the binding sites for the lactose ligand which can specifically bind with Gal-1. The Cu2O@Au nanocomposites provide the synergic effect of electrochemical signal amplification and lactose–galectin reaction as the recognition strategy. Under optimal conditions, the proposed biosensor exhibits a variation of electrochemical responses to different concentrations of Gal-1 ranging from 0.1 pg ml−1 to 10 ng ml−1. This work presents an alternative electrochemical biosensor for the detection of tumor biomarkers based on a simple and economical lactose ligand incorporated Cu2O@Au biosensor platform.

Interference-free and high precision biosensor based on surface enhanced Raman spectroscopy integrated with surface molecularly imprinted polymer technology for tumor biomarker detection in human blood

The reliable quantitative analysis of tumor biomarkers in circulating blood is crucial for cancer early screening, therapy monitoring and prognostic prediction. Herein, a novel biosensor combing surface-enhanced Raman spectroscopy (SERS) and surface molecularly imprinted polymer (SMIP) technology was developed for quantitative detection of carcinoembryonic antigen (CEA) that is closely related to several common cancers. Owing to the use of SMIP, recognition sites with high affinity to the target of interest can be well imprinted on the surface of SERS substrate, leading to a more stable and specific capture ability. In addition, two layers of core-shell nanoparticles were integrated to this SERS substrate to form highly efficient electromagnetic enhancement for SERS measurement via the generation of lots of “hot spot”. Besides, a unique Raman reporter (CC) with silent Raman signals at 2024 cm−1 was capsulated in the nanoparticles to avoid the optical noises originating from endogenous molecules at fingerprint region (300-1800 cm−1). Meanwhile, we employed an internal standard molecular (CN) to real time correct the fluctuating signals of Raman reporter when performing the quantitative analysis. Due to these features, a limit of detection (LOD) of 0.064 pg mL−1 with the detection range of 0.1 pg mL−1 - 10 μg mL−1 can be achieved by this assay. Excitingly, this technology even showed wonderful performances for CEA detection in real blood from cancer patients, demonstrating great potential for biomarker-based cancer screening.

Dendritic core-shell rhodium@platinum-cobalt nanocrystals for ultrasensitive electrochemical immunoassay of squamous cell carcinoma antigen

Squamous cell carcinoma antigen (SCCA) has great diagnostic values for its high specificity (90–96%) in diagnosis of squamous cell carcinoma especially cancers, where electrochemical immunoassay is powerful for its accurate and reliable detection of specific tumor biomarkers at very low levels. In this work, well-defined three-dimensional dendritic core-shell rhodium@platinum-cobalt nanocrystals (Rh@PtCo NCs) were facilely prepared via a simple one-pot solvothermal strategy. Taking advantage of dramatically enhanced catalytic activity of the Rh@PtCo NCs for catechol oxidation, a label-free electrochemical immunosensor was developed for highly sensitive assay of SCCA. Under optimal conditions, the electrochemical signals were linearly correlated with the logarithm of the SCCA concentration, showing a broad linear range from 0.0001 to 10.0 ng mL−1 and a ultralow detection limit of 0.04 pg mL−1 (S/N = 3). The resultant immunosensor was further exploited for actual analysis of serum samples with convinced results. This immunosensor has good expectation in actual samples analysis and provides a universal approach for detection of other tumor markers and disease surveillance.

7Hsp70 serum levels in pet dogs\u2014a potential diagnostic biomarker for spontaneous round cell tumors

The concentration of circulating heat shock protein 70 (Hsp70) was measured in liquid biopsies of canine tumor patients as a potential biomarker. Compared with rodent tumor models, spontaneously occurring tumors in pet dogs reflect the clinical situation of human patients better, as dogs cohabitate with their owners in the same environment, reach a much older age than rodents, can provide blood samples much more frequently, and receive up-to-date medical care and, similar to humans, their tumors show a high genetic heterogeneity. Due to the species-specific sequence homology of human and canine Hsp70, two human enzyme-linked immunosorbent assay (ELISA) systems (R&D and lipHsp70) were used to measure canine Hsp70 concentrations in serum and plasma. In general, higher Hsp70 concentrations were found in serum compared with plasma samples of dogs, and the lipHsp70 ELISA detected higher peak concentrations of Hsp70 in a broader range than the R&D ELISA. Compared with a tumor-free control group, serum Hsp70 concentrations were higher in tumor-bearing dogs, irrespective of breed, age, body weight, and gender. A sub-classification of the different tumors according to their cytological characteristics revealed significantly elevated Hsp70 serum concentrations in dogs with round cell tumors (p < 0.01), a heterogeneous group of malignancies with hematopoietic origin such as mast cells, plasma cells, lymphocytes, histiocytes, and melanomas. Future studies with larger patient cohorts and well-defined tumor sizes are necessary to elucidate the role of serum Hsp70 as a biomarker for tumor detection and monitoring of outcome in pet animals.