Analysis Of Cancer Biomarkers Research Articles

A magnetic bead array-based fluorescence immunoassay platform for robust cancer biomarker analysis in clinical blood samples

A magnetic bead array-based fluorescence immunoassay platform for robust cancer biomarker analysis in clinical blood samples

Computational Ensemble Gene Co-Expression Networks for the Analysis of Cancer Biomarkers

Gene networks have become a powerful tool for the comprehensive examination of gene expression patterns. Thanks to these networks generated by means of inference algorithms, it is possible to study different biological processes and even identify new biomarkers for such diseases. These biomarkers are essential for the discovery of new treatments for genetic diseases such as cancer. In this work, we introduce an algorithm for genetic network inference based on an ensemble method that improves the robustness of the results by combining two main steps: first, the evaluation of the relationship between pairs of genes using three different co-expression measures, and, subsequently, a voting strategy. The utility of this approach was demonstrated by applying it to a human dataset encompassing breast and prostate cancer-associated stromal cells. Two gene networks were computed using microarray data, one for breast cancer and one for prostate cancer. The results obtained revealed, on the one hand, distinct stromal cell behaviors in breast and prostate cancer and, on the other hand, a list of potential biomarkers for both diseases. In the case of breast tumor, ST6GAL2, RIPOR3, COL5A1, and DEPDC7 were found, and in the case of prostate tumor, the genes were GATA6-AS1, ARFGEF3, PRR15L, and APBA2. These results demonstrate the usefulness of the ensemble method in the field of biomarker discovery.

Near-infrared light-assisted dual-modal photoelectrochemical and photo-fuel-cell-driven self-powered biosensor for detection of miRNA-21 in living cancer cells

The advancement of the dual-modal photoelectrochemical (PEC) and photo-fuel-cell-driven self-powered biosensor (PFC-SPB) with the aid of near-infrared light (NIR) irradiation can present a robust sensing strategy for the feasible, sensitive detection of cancer biomarkers in living cancer cells. In this work, a novel NIR-assisted dual-modal PEC and PFC-SPB is proposed for the sensitive inspection of miRNA-21 based on a 2D/2D p-n heterojunction, which is prepared by the in-situ growth of zinc indium sulfide nanosheets (ZnIn2S4 NSs) over the 2D zinc porphyrin-based metal–organic frameworks using meso-tetra (4-carboxyphenyl) porphyrin as ligand (denoted as ZnIn2S4@Zn-TCPP). Benefiting the integration of the features of ZnIn2S4 and Zn-TCPP, the developed p-n heterojunction demonstrates rich element vacancies, regulated NIR absorption, and improved charge separation efficiency of the photogenerated electron–hole pairs. Thus, the ZnIn2S4@Zn-TCPP photoelectrode affords the boosted photoelectric conversion efficiency and large output voltage as well as large amounts of anchored complementary DNA strand due to the comprehensive interaction. The constructed dual-modal PEC and PFC-SPB, respectively, exhibit an ultralow limit of detection (LOD) of 0.43 and 0.71 fM toward miRNA-21 within a range of 1 fM to 1 nM, along with the other superior sensing abilities. The PFC-driven self-powered biosensor also shows a great potential in the selective detection of miRNA-21 contained in living cancer cells. The present work provides an NIR-assisted dual-modal biosensing strategy for the efficient analysis of cancer biomarkers in vitro.

Fascinating Immobilization-Free Electrochemical Immunosensing Strategy Based on the Cooperation of Buoyancy and Magnetism.

Rapid and accurate detection of biomolecules is of vital importance for the diagnosis of disease and for performing timely treatments. The point-of-care analysis of cancer biomarkers in the blood with low cost and easy processing is still challenging. Herein, an advanced and robust strategy, which integrates the buoyant recognition probe with the magnetic reporter probe in one solution, was first proposed for immobilization-free electrochemical immunosensing. The tumor marker of alpha fetoprotein (AFP) can be captured immune-buoyantly, and then a multifunctional magnetic reporter probe in pseudo-homogeneous solution was further captured to fulfill a sandwich-type immunoreaction. The residual magnetic reporter probe can be firmly and efficiently attracted on a magnetic glassy carbon electrode to fulfill the conversion of the target AFP amount into the residual magnetic electrochemical signal indicator. As a result, the electrochemical signal of methylene blue can accurately reflect the original level of target antigen AFP concentration. By integrating buoyancy-driven quasi-homogenous biorecognition with magnetism-mediated amplification and signal output, the proposed immobilization-free electrochemical immunosensing strategy displayed a wide range of linear response (100 fg mL-1 to 10 ng mL-1), low detection limit (14.52 fg mL-1), and good reproducibility, selectivity, and stability. The designed strategy manifests remarkable advantages including assay simplicity, rapidness, and high sensitivity owing to the in-solution instead of on-electrode biorecognition that could accelerate and improve the biorecognition efficiency. To the best of our knowledge, this is the first cooperation of buoyancy-driven biorecognition with magnetism-mediated signal output in bioanalysis, which would be attractive for rapid clinic biomedical application. Thus, this work provides a fresh perspective for convenient and favorable immobilization-free electrochemical biosensing of universal biomolecules.

Extracellular Vesicles in Liquid Biopsies as Biomarkers for Solid Tumors

Extracellular vesicles (EVs) are secreted by all living cells and are ubiquitous in every human body fluid. They are quite heterogeneous with regard to biogenesis, size, and composition, yet always reflect their parental cells with their cell-of-origin specific cargo loading. Since numerous studies have demonstrated that EV-associated proteins, nucleic acids, lipids, and metabolites can represent malignant phenotypes in cancer patients, EVs are increasingly being discussed as valuable carriers of cancer biomarkers in liquid biopsy samples. However, the lack of standardized and clinically feasible protocols for EV purification and characterization still limits the applicability of EV-based cancer biomarker analysis. This review first provides an overview of current EV isolation and characterization techniques that can be used to exploit patient-derived body fluids for biomarker quantification assays. Secondly, it outlines promising tumor-specific EV biomarkers relevant for cancer diagnosis, disease monitoring, and the prediction of cancer progression and therapy resistance. Finally, we summarize the advantages and current limitations of using EVs in liquid biopsy with a prospective view on strategies for the ongoing clinical implementation of EV-based biomarker screenings.

Toward Personalized Nanomedicine: The Critical Evaluation of Micro and Nanodevices for Cancer Biomarker Analysis in Liquid Biopsy.

Liquid biopsy for the analysis of circulating cancer biomarkers (CBs) is a major advancement toward the early detection of cancer. In comparison to tissue biopsy techniques, liquid biopsy is relatively painless, offering multiple sampling opportunities across easily accessible bodily fluids such as blood, urine, and saliva. Liquid biopsy is also relatively inexpensive and simple, avoiding the requirement for specialized laboratory equipment or trained medical staff. Major advances in the field of liquid biopsy are attributed largely to developments in nanotechnology and microfabrication that enables the creation of highly precise chip-based platforms. These devices can overcome detection limitations of an individual biomarker by detecting multiple markers simultaneously on the same chip, or by featuring integrated and combined target separation techniques. In this review, the major advances in the field of portable and semi-portable micro, nano, and multiplexed platforms for CB detection for the early diagnosis of cancer are highlighted. A comparative discussion is also provided, noting merits and drawbacks of the platforms, especially in terms of portability. Finally, key challenges toward device portability and possible solutions, as well as discussing the future direction of the field are highlighted.

Integrated "lab-on-a-chip" microfluidic systems for isolation, enrichment, and analysis of cancer biomarkers.

The liquid biopsy has garnered considerable attention as a complementary clinical tool for the early detection, molecular characterization and monitoring of cancer over the past decade. In contrast to traditional solid biopsy techniques, liquid biopsy offers a less invasive and safer alternative for routine cancer screening. Recent advances in microfluidic technologies have enabled handling of liquid biopsy-derived biomarkers with high sensitivity, throughput, and convenience. The integration of these multi-functional microfluidic technologies into a 'lab-on-a-chip' offers a powerful solution for processing and analyzing samples on a single platform, thereby reducing the complexity, bio-analyte loss and cross-contamination associated with multiple handling and transfer steps in more conventional benchtop workflows. This review critically addresses recent developments in integrated microfluidic technologies for cancer detection, highlighting isolation, enrichment, and analysis strategies for three important sub-types of cancer biomarkers: circulating tumor cells, circulating tumor DNA and exosomes. We first discuss the unique characteristics and advantages of the various lab-on-a-chip technologies developed to operate on each biomarker subtype. This is then followed by a discussion on the challenges and opportunities in the field of integrated systems for cancer detection. Ultimately, integrated microfluidic platforms form the core of a new class of point-of-care diagnostic tools by virtue of their ease-of-operation, portability and high sensitivity. Widespread availability of such tools could potentially result in more frequent and convenient screening for early signs of cancer at clinical labs or primary care offices.

An all-graphdiyne electrochemiluminescence biosensor for the ultrasensitive detection of microRNA-21 based on target recycling with DNA cascade reaction for signal amplification.

Graphdiyne oxide quantum dots (GDYO QDs), as derivatives of graphdiyne (GDY), have excellent electroconductibility and luminous properties and can be applied as a new ECL emitter. Herein, an electrochemiluminescence (ECL) biosensor for miRNA-21 ultrasensitive determination is constructed based on AuNPs/GDY, GDYO QD and oligonucleotide signal amplification strategy that integrates DNA walker and hybridization chain reaction (HCR) amplification. As electrode substrate material, AuNPs/GDY can not only bond with the aptamer CP but can also enhance the conductivity of the interface. When miRNA-21 exists, the DNA walker process is initiated, and the signaling probes are introduced on the electrode surface, producing abundant double-stranded H1/H2; then, H3/H4 undergoes complementary base pairing with H1/H2 through HCR. With the increase in miRNA-21, the 3D DNA nanomachine is actively manipulated, resulting in a gradual increase in ECL signal. This ECL biosensor demonstrates outstanding performance in the determination of miRNA-21 in the linear range from 0.1 fM to 1 nM. This study offers a new sensitive idea for the clinical analysis of cancer biomarkers.

Breast cancer molecular diagnostics in Rwanda: a cost-minimization study of immunohistochemistry versus a novel GeneXpert® mRNA expression assay.

To compare the financial and time cost of breast cancer biomarker analysis by immunohistochemistry with that by the Xpert® STRAT4 assay. We estimated costs (personnel, location, consumables and indirect) and time involved in breast cancer diagnosis at the Butaro Cancer Centre of Excellence, Rwanda, using time-driven activity-based costing. We performed a cost-minimization analysis to compare the cost of biomarker analysis for estrogen receptor, progesterone receptor and human epidermal growth factor receptor-2 status with immunohistochemistry versus STRAT4. We performed sensitivity analyses by altering laboratory-specific parameters for the two methods. We estimated that breast cancer diagnosis in Rwanda costs 138.29 United States dollars (US$) per patient when conducting biomarker analysis by immunohistochemistry. At a realistic immunohistochemistry antibody utilization efficiency of 70%, biomarker analysis comprises 48.7% (US$67.33) of diagnostic costs and takes 33min. We determined that biomarker analysis with STRAT4 yields a reduction in diagnosis cost of US$7.33 (10.9%; 7.33/67.33), and in pathologist and technician time of 20min (60.6%; 20/33), per patient. Our sensitivity analysis revealed that no cost savings would be made in laboratories with antibody utilization efficiencies over 90%, or where only estrogen and/or progesterone receptor status are assessed; however, such operational efficiencies are unlikely, and more laboratories are pursuing human epidermal growth factor receptor-2 analysis as targeted therapies become increasingly available. Breast cancer biomarker analysis with STRAT4 has the potential to reduce the required human and capital resources in sub-Saharan African laboratories, leading to improved treatment selection and better clinical outcomes.

Microbiome and Metabolomics in Liver Cancer: Scientific Technology.

Primary liver cancer is a heterogeneous disease. Liver cancer metabolism includes both the reprogramming of intracellular metabolism to enable cancer cells to proliferate inappropriately and adapt to the tumor microenvironment and fluctuations in regular tissue metabolism. Currently, metabolomics and metabolite profiling in liver cirrhosis, liver cancer, and hepatocellular carcinoma (HCC) have been in the spotlight in terms of cancer diagnosis, monitoring, and therapy. Metabolomics is the global analysis of small molecules, chemicals, and metabolites. Metabolomics technologies can provide critical information about the liver cancer state. Here, we review how liver cirrhosis, liver cancer, and HCC therapies interact with metabolism at the cellular and systemic levels. An overview of liver metabolomics is provided, with a focus on currently available technologies and how they have been used in clinical and translational research. We also list scalable methods, including chemometrics, followed by pathway processing in liver cancer. We conclude that important drivers of metabolomics science and scientific technologies are novel therapeutic tools and liver cancer biomarker analysis.

Cancer and Disease Diagnosis - Biosensor as Potential Diagnostic Tool for Biomarker Detection.

Analysis of cancer biomarkers has enormous promise for advancing our molecular understanding of illness and facilitating more precise and timely diagnosis and follow-up care. MicroRNA, exosomes, ctDNA, CTCs, and proteins are only some of the circulating biomarkers that can be detected by liquid biopsy instead of the more intrusive and time-consuming process of doing a tissue biopsy. As the cancer diagnosis bio-markers reveal ultra-low levels in the early stages of the disease, highly sensitive approaches are urgently required. Researchers have taken an interest in a optical biosensor for detecting cancer biomarkers as a potential tool for early disease diagnosis. These techniques have the potential to aid in the development of effective treatments, ultimately leading to a higher rate of patient survival. This review briefly discuss the i) understanding of cancer and biomarkers for early diagonosis purpose ii) Molecular methods and ii) biosensor-based diagnostics. The reseach primary focus on advancement in biosensor design using various concepts ie., Electrochemical, Chemiluminescence and Colorimetric, Surface plasmons (SP), Surface plasmon resonance (SPR), localized surface plasmon resonance (LSPR), Fluorescence, Fiber-based sensors, Terahertz based biosensors, and Surface enhanced Raman spectroscopy (SERS). As a result of the local electric field amplification around plasmonic (usually gold and silver) nanostructures, surface-enhanced Raman spectroscopy (SERS) has emerged as a rapid, selective, and sensitive alternative to conventional laboratory analytical methods, making significant strides in a number of biosensing applications but still under developing stage to be used as diagnostic tool in clinical research.

Core Concept Of Bioelectronic Medicine And Their Theranostic Application In Cancer

Bioelectronic medicine is a way of treating in which the electrical communication networks of the body are altered to treat disease. Bioelectric circuitry in every cell creates a resting membrane potential as well as endogenous electric fields that regulate cell function and communication. The theory that cancer is caused by bioelectrical circuitry malfunctions in cells is now widely acknowledged. This opens up the possibility of using bioelectronic medicine to develop new cancer treatments by influencing the disease's bioelectrical properties. To meet societal and technical obstacles, today's individuals aspire to better their quality of life. So, in the midst of all of this, bioelectronic medicine has altered the way we practice medicine and has proven to dramatically enhance healthcare outcomes. This technique employs a variety of procedures, such as neuromodulation, to treat a specific disease, ailment, or damage. Interfacing electronics with nerves to specifically target the biological processes underlying diseases. In cancer, screening and early detection are crucial for the patient's therapy to be successful and for the patient's survival percentage to grow. For illness diagnosis, therapy, and follow-up, the development of analytical methods for non-invasive detection through the analysis of cancer biomarkers is crucial. Tumor biomarkers are substances or processes that indicate the existence of cancer in the body in clinical situations. These biomarkers can be detected using biosensors, which are popular alternatives to traditional approaches due to their speed, accuracy, and point-of-care applicability. Here we have discussed the consequences of current cancer treatments and bioelectronic medicine a combat for cancer diagnosis and treatment.

Fabrication of a new SPME fiber based on Polyacrylic acid/ MIL-88(Fe)-NH2 composite as a self-healing coating for the analysis of breast cancer biomarkers in the urine sample

Analysis of cancer biomarkers in the body fluids is a new method for early detection of illness. However, due to the complex matrices of samples, the application of a pre-treatment method is unavoidable before the final analysis by gas chromatography (GC). Solid-phase microextraction (SPME) is a simple and, promising pre-concentration and separation method that its coatings are modified with different materials on the fibers. A new innovative self-healing polyacrylic acid PAA/MIL-88(Fe)-NH2 composite was synthesized as an SPME coating. The parameters including pH, crosslinker, and MIL-88(Fe)-NH2 content were optimized to formulate the composite. The prepared fiber was used to extract 2-pentanone, 2-heptanone, ethyl propionate, p-xylene, 1,2,4-trimethylbenzene, and o-cresol as a biomarker in breast cancer from urine samples. The prepared PAA/MIL-88(Fe)-NH2 SPME fibers demonstrate excellent repeatability (relative standard deviation (RSD%)< 3.4%) and satisfactory reproducibility (RSD%<6.9%). The central composite design method was applied for the optimization of extraction parameters. Under the optimum conditions, linear dynamic ranges for biomarkers were in the acceptable range with correlation coefficients higher than 0.98. The detection limits of them were less down 0.0016 µg L−1. Self-healing ability of fiber coating increased useful lifetime (about 120 times extraction with one fiber) as well as accuracy, reproducibility, and recovery of fibers.

Gastric cancer biomarker analysis in patients treated with different adjuvant chemotherapy regimens within SAMIT, a phase III randomized controlled trial

Biomarkers for selecting gastric cancer (GC) patients likely to benefit from sequential paclitaxel treatment followed by fluorinated-pyrimidine-based adjuvant chemotherapy (sequential paclitaxel) were investigated using tissue samples of patients recruited into SAMIT, a phase III randomized controlled trial. Total RNA was extracted from 556 GC resection samples. The expression of 105 genes was quantified using real-time PCR. Genes predicting the benefit of sequential paclitaxel on overall survival, disease-free survival, and cumulative incidence of relapse were identified based on the ranking of p-values associated with the interaction between the biomarker and sequential paclitaxel or monotherapy groups. Low VSNL1 and CD44 expression predicted the benefit of sequential paclitaxel treatment for all three endpoints. Patients with combined low expression of both genes benefitted most from sequential paclitaxel therapy (hazard ratio = 0.48 [95% confidence interval, 0.30–0.78]; p < 0.01; interaction p-value < 0.01). This is the first study to identify VSNL1 and CD44 RNA expression levels as biomarkers for selecting GC patients that are likely to benefit from sequential paclitaxel treatment followed by fluorinated-pyrimidine-based adjuvant chemotherapy. Our findings may facilitate clinical trials on biomarker-oriented postoperative adjuvant chemotherapy for patients with locally advanced GC.

MALDI imaging mass spectrometry mapping of the glycocalyx

The glycocalyx is a dense complex matrix of glycoproteins, glycosaminoglycans and glycolipids on the cell surface involved in maintaining cell integrity and interactions with the cellular microenvironment. Alterations to the glycan constituents of the glycocalyx are implicit in progression of diseases like cancer, but its overall complexity makes biomolecular analysis very challenging. Changes in cell surface protein glycosylation are common alterations that occur with tumor progression and reflect the clinical diagnostic detection of many cancer biomarkers like prostate specific antigen and CA19‐9. There is also an underlying metabolic connection to cancer glycosylation changes and the well described changes in tumor metabolism linked with glucose and glutamine utilization. Structurally, these potential tumor glycans are represented by multiple sialylated and fucosylated species including known tumor antigens such as sialyl Lewis X, sialyl Lewis A and Lewis Y motifs. Imaging mass spectrometry of clinical tissues and new enzymatic and chemical sample preparation strategies have provided new approaches for spatial characterization of the glycocalyx constituents. N‐glycan MALDI imaging MS allows direct histopathology localization of these glycans to tumor, stroma and immune regions. Peptide N‐glycosidase and other glycosidases are sprayed directly on tissues, and released glycans are detected spatially on a MALDI mass spectrometer. The method was developed to work on clinical formalin‐fixed paraffin embedded tissue slides readily obtainable from pathology services. Generally, 45‐60 N‐glycan species are detected in most tissues, and these numbers can increase to 100 or more in advanced cancer tissues. Example data from prostate, colon, and pancreatic cancer tissues will be presented, including new workflows that specifically target identification of isomeric species of branched fucosylated and sialylated N‐glycans. This includes combined use of immunohistochemistry staining of known carbohydrate tumor antigens prior to N‐glycan imaging, performed on the same slide. Strategies to use the tissue N‐glycan distribution maps for glycoproteomic and cancer biomarker analysis will also be presented. Additional glycosidases, other enzymes and other imaging modalities can also be incorporated into the established workflows to better define the complex organization of the tissue glycocalyx.

Monosaccharide-mediated rational synthesis of a universal plasmonic platform with broad spectral fluorescence enhancement for high-sensitivity cancer biomarker analysis

BackgroundEffective and accurate screening of oncological biomarkers in peripheral blood circulation plays an increasingly vital role in diagnosis and prognosis. High-sensitivity assays can effectively aid clinical decision-making and intervene in cancer in a localized status before they metastasize and become unmanageable. Meanwhile, it is equally pivotal to prevent overdiagnosis of non-life-threatening cancer by eliminating unnecessary treatment and repeated blood draws. Unfortunately, current clinical screening methodologies can hardly simultaneously attain sufficient sensitivity and specificity, especially under resource-restrained circumstances. To circumvent such limitations, particularly for cancer biomarkers from early-onset and recurrence, we aim to develop a universal plasmonic platform for clinical applications, which macroscopically amplifies multiplexed fluorescence signals in a broad spectral window and readily adapts to current assay setups without sophisticated accessories or expertise at low cost.MethodsThe plasmonic substrate was chemically synthesized in situ at the solid–liquid interface by rationally screening a panel of reducing monosaccharides and tuning the redox reactions at various catalyst densities and precursor concentrations. The redox properties were studied by Benedict’s assay and electrochemistry. We systemically characterized the morphologies and optical properties of the engineered plasmonic Ag structures by scanning electron microscopy (SEM) and spectroscopy. The structure-fluorescence enhancement correlation was explicitly explained by the finite-difference time-domain (FDTD) simulation and a computational model for gap distribution. Next, we established an enhanced fluoroimmunoassay (eFIA) using a model biomarker for prostate cancer (PCa) and validated it in healthy and PCa cohorts. Prognosis was explored in patients subject to surgical and hormonal interventions following recommended PCa guidelines.ResultsThe monosaccharide-mediated redox reaction yielded a broad category of Ag structures, including sparsely dispersed nanoparticles (NPs) of various sizes, semi-continuous nanoislands, and crackless continuous films. Optimal broad-spectral fluorescence enhancement from green to far-red was observed for the inhomogeneous, irregularly-shaped semi-continuous Ag nanoisland substrate (AgNIS), synthesized from a well-balanced redox reaction at a stable rate mediated by mannose. In addition, different local electric field intensity distributions in response to various incident excitations were observed at the nanoscale, elucidating the need for irregular and inhomogeneous structures. AgNIS enabled a maximized 54.7-fold macroscopically amplified fluorescence and long-lasting photostability. Point-of-care availability was fulfilled using a customized smartphone prototype with well-paired optics. The eFIA effectively detected the PCa marker in cell lines, xenograft tumors, and patient sera. The plasmonic platform rendered a diagnostic sensitivity of 86.0% and a specificity of 94.7% and capably staged high-grade PCa that the clinical gold standard test failed to stratify. Patient prognosis of robotic-assisted surgeries and hormone therapies was non-invasively monitored following efficient medical interventions. The assay time was significantly curtailed on the plasmonic platform upon microwave irradiation.ConclusionsBy investigating the effects of reducing monosaccharides on the seed-mediated chemical synthesis of plasmonic Ag structures, we deduced that potent multiplexed fluorescence enhancement originated from both an adequate reducing power and a steady reduction rate. Furthermore, the inhomogeneous structure with adequate medium gap distances afforded optimal multiwavelength fluorescence enhancement, thus empowering an effective eFIA for PCa. The clinically validated diagnostic and prognostic features, along with the low sample volume, point-of-care feasibility with a smartphone, and microwave-shortened assay time, warrant its potential clinical translation for widespread cancer biomarker analysis.Graphical

Discordant prognosis of mismatch repair deficiency in colorectal and endometrial cancer reflects variation in antitumour immune response and immune escape.

Defective DNA mismatch repair (dMMR) causes elevated tumour mutational burden (TMB) and microsatellite instability (MSI) in multiple cancer types. dMMR/MSI colorectal cancers (CRCs) have enhanced T‐cell infiltrate and favourable outcome; however, this association has not been reliably detected in other tumour types, including endometrial cancer (EC). We sought to confirm this and explore the underpinning mechanisms. We first meta‐analysed CRC and EC trials that have examined the prognostic value of dMMR/MSI and confirmed that dMMR/MSI predicts better prognosis in CRC, but not EC, with statistically significant variation between cancers (hazard ratio [HR] = 0.63, 95% confidence interval [CI] = 0.54–0.73 versus HR = 1.15, 95% CI = 0.72–1.58; P INT = 0.02). Next, we studied intratumoural immune infiltrate in CRCs and ECs of defined MMR status and found that while dMMR was associated with increased density of tumour‐infiltrating CD3+ and CD8+ T‐cells in both cancer types, the increases were substantially greater in CRC and significant only in this group (P INT = 4.3e‐04 and 7.3e‐03, respectively). Analysis of CRC and EC from the independent Cancer Genome Atlas (TCGA) series revealed similar variation and significant interactions in proportions of tumour‐infiltrating lymphocytes, CD8+, CD4+, NK cells and immune checkpoint expression, confirming a more vigorous immune response to dMMR/MSI in CRC than EC. Agnostic analysis identified the IFNγ pathway activity as strongly upregulated by dMMR/MSI in CRC, but downregulated in EC by frequent JAK1 mutations, the impact of which on IFNγ response was confirmed by functional analyses. Collectively, our results confirm the discordant prognosis of dMMR/MSI in CRC and EC and suggest that this relates to differences in intratumoural immune infiltrate and tumour genome. Our study underscores the need for tissue‐specific analysis of cancer biomarkers and may help inform immunotherapy use. © 2022 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of The Pathological Society of Great Britain and Ireland.

An Electrochemical and Raman Scattering Dual Detection Biosensor for Rapid Screening and Biomolecular Profiling of Cancer Biomarkers

Detecting circulating biomarkers sensitively and quantitatively is paramount for cancer screening, diagnosis, and treatment selection. Particularly, screening of a panel of circulating protein biomarkers followed by mapping of individual biomarkers could assist better diagnosis and understanding of the cancer progression mechanisms. Herein, we present a miniaturized biosensing platform with dual readout schemes (electrochemical and Surface enhanced Raman scattering (SERS)) for rapid cancer screening and specific biomarker expressional profiling to support cancer management. Our approach utilizes a controlled nanomixing phenomena under alternative current electrohydrodynamic condition to improve the isolation of cancer-associated circulating proteins (i.e., Epidermal growth factor receptor (EGFR), BRAF, Programmed death-ligand 1 (PD-L1)) with antibody functionalized sensor surface for rapid and efficient isolation of the targets and subsequent labelling with SERS nanotags. The method employs Differential Pulse Voltammetry (DPV) for rapidly screening for the presence of the circulating proteins on biosensor surface irrespective of their type. Upon positive DPV detection, SERS is applied for sensitive read-out of individual biomarkers biomarker levels. In a proof-of-concept study, we demonstrate the dual detection biosensor for analysing circulating BRAF, EGFR and PDL-1 proteins and successfully screened both ensemble and individual biomarker expressional levels as low as 10 pg (1 ng/mL). Our findings clearly indicate the potential of the proposed method for cancer biomarker analysis which may drive the translation of this dual sensing concept in clinical settings.

Membrane Protein and Extracellular Acid Heterogeneity-Driven Amplified DNA Logic Gate Enables Accurate and Sensitive Identification of Cancer Cells.

DNA logic gates, as a class of smart molecular devices with excellent biocompatibility and convenient information processing mode, have been widely used for identification of cancer cells based on logic analysis of cancer biomarkers. However, most of the developed DNA logic gates for identification of cancer cells are mainly driven by homogeneous biomarkers such as membrane proteins or RNAs, which may suffer from insufficient accuracy. Herein, we reported a membrane protein and extracellular acid heterogeneity-driven amplified DNA logic gate (HDLG) for accurate and sensitive identification of cancer cells by combining the superior signal amplification characteristics of the hybridization chain reaction (HCR) and the precise computation ability of the logic operation. In this strategy, a DNA aptamer was employed for membrane protein recognition, and a split i-motif was used for the response of the extracellular acid. Only when the two heterogeneous biomarkers existed simultaneously, the DNA logic gate could be driven to perform the "AND" logic operation and induce the formation of an intact trigger to initiate a HCR process on the cell surface, generating an amplified "ON" fluorescence signal. Benefiting from the design of heterogeneity-driven and signal amplification, this DNA logic gate could not only autonomously perform high-resolution fluorescence imaging on the surface of target cancer cells, but also perform sensitive analysis of target cancer cells with a cell number of 70 detected in 200 μL of buffer and desirable accuracy in differentiating target cancer cells from complicated cell mixtures. We anticipate that this novel HDLG is expected to be applied in precise disease diagnosis.

Impact of GC-MS in Analysis of Cancer Biomarkers

Cancer is the one of the most dreadful diseases of the modern age. A huge number of chemical biomarkers are associated with progression and abnormalities...