Cancer Therapy Monitoring Research Articles

EMT in breast cancer metastasis: an interplay of microRNAs, signaling pathways and circulating tumor cells.

Epithelial to Mesenchymal Transition (EMT) is a biological process characterized by the transition from immotile epithelial cells to motile mesenchymal cells. Though shown to be implicated in many biological processes, it has also been identified to enhance migration and invasion of cancer cells leading to metastasis. A class of microRNAs called "oncomiRs" plays a significant role in the regulation of malignant transformation and metastasis. In this review, the ability of different signaling pathways in controlling EMT through well-defined regulatory networks, and the role exerted by oncomiRs in regulating the specific signaling pathways like TGF-β, Wnt, Notch and Hedgehog in modulating breast cancer metastasis have been discussed with updated information. Further, this review focuses on the significance of up and down regulated microRNAs in the pathogenesis and progression of breast cancer and how such microRNAs could be treated as potential therapeutic targets to circumvent cancer. As a prospective strategy, we highlight the importance of circulating tumor cells (CTCs) and their derived microRNAs as prognostic indicators and cancer therapy monitoring tools.

Engineering magnetic nanoparticles and their integration with microfluidics for cell isolation

Isolation of cancer cells, bacteria, and viruses from peripheral blood has important applications in cancer diagnosis, therapy monitoring, and drug development. Magnetic particles functionalized with antibodies that target receptors of cancer cells have been shown to isolate such entities using magnetic field gradients. Here, we report enhancement in capture efficiency and specificity by engineering magnetic nanoparticles and integrating them with microfluidics for the enumeration of tumor cells. Nanoparticles were made from iron oxide, coated with poly(ethylene glycol), and conjugated through avidin-biotin chemistry with antibody specifically against epithelial cell adhesion molecule (EpCAM). On exposure of targeted nanoparticles to tumor cells, specific uptake by EpCAM-expressing tumor cells (e.g., BxPC3, a pancreatic cancer cell) was observed, whereas there was negligible uptake by cells with low EpCAM expression (e.g., CCRF-CEM, a leukemia cell). Using an arrangement of magnets called a Halbach array, capture efficiency and specificity towards BxPC3 cells tagged with magnetic nanoparticles were enhanced, compared to conditions without the magnetic field gradient and/or without magnetic nanoparticles, either in buffer or in whole blood. These results illustrate that engineered magnetic nanoparticles and their integration with microfluidics have great potential for tumor cell enumeration and cancer prognosis.

Integrated Microfluidic Device for Accurate Extracellular Vesicle Quantification and Protein Markers Analysis Directly from Human Whole Blood.

Extracellular vesicles (EVs) have the potential to be utilized as disease-specific biomarkers. Although strategies for on-chip isolation and detection of EVs have recently been developed, they need preprocessing of clinical samples and are not accurate enough for disease diagnosis just judging by EVs concentration. Here, we designed an integrated microfluidic device named a plasma separation and EV detection (PS-ED) chip for plasma separation, quantification, and high-throughput protein analysis of EVs directly from clinical whole blood samples. The device included two modules (PS and ED module): the PS module was a six-loop microchannel for rapid separation of plasma from clinical whole blood samples under inertial force; the amount of EVs in the separated plasma kept the same value as in the initial blood samples. The module reduced the mechanical damage to the blood cells and thus reduced the interference of debris or cellular contents from damaged cells during EVs detection; the ED module contained four S-channels for quantification and high-throughput protein analysis of EVs; a wide detection range from 2.5 × 102 to 2.5 × 108 particles/μL with a detection limit of 95 particles/μL was obtained. Through simultanously monitoring three proteins (CD81, CD24, and EpCAM) of EVs, the cancer type can be accurately confirmed. Furthermore, clinical blood sample analysis verified that the proposed device could be used for accurate diagnosis and therapy monitoring of ovarian cancer.

Volatile Compounds Are Involved in Cellular Crosstalk and Upregulation.

Cell-cell cross talk is of great importance in cancer research due to its major role in proliferation, differentiation, migration, and influence on the apoptotic pathway. Different cell-cell communication mechanisms have come mainly from proteomic and genomic approaches. In this paper, a new route is reported for cross talk between cancer cells that occurs, even when they are far away from each other. Single-cell and culture analysis shows that upregulation of cancer cells emits hundreds of volatile organic compounds (VOCs) into their headspace. Part of the VOCs remains without any change, disregarding the biological environment around it. The other part of the VOCs is exchanged between monocultures of the cells as well as between co-cultures of the cells with no physical contact between them, leading to different changes in growth than when left on their own. The chemical nature and composition of these VOCs have been determined and are discussed herein. Cell-to-cell cross talk has the advantage of being suitable for transfer/diffusion over relatively long distances. It would thus be expected to serve as a shuttling pad toward the development of advanced approaches that could enable very early detection of cancer and/or monitoring of metastasis and related cancer therapy.

Parallel Comparison of Cancer Antigen 15.3 and Cancer Antigen 27.29 in Breast Cancer

Abstract Objectives Mucins are large and complex glycoproteins expressed by several epithelial tissues. The expression of MUC1 mucin is upregulated in breast cancer and the protein is released in high amounts in patients’ blood. Cancer antigens (CA) 15.3 and CA 27.29, which measure different epitopes of MUC1 protein, are useful biomarkers for postoperative surveillance and therapy monitoring of advanced breast cancer. The aim of this study was to compare CA 15.3 assay and CA 27.29 assay by using paired patient samples. Methods The study was performed in 523 patient samples. CA 15.3 was measured by a sandwich assay on Beckman Coulter UniCel® DxI 800. CA 27. 29 was measured by a competitive immunoassay on Siemens ADVIA Centaur System. Results On Deming regression, the slope was 0.310 with an intercept of 8.6 and correlation coefficient of 0.951. The mean bias between the two assays was –39.2 (–56.6%). The agreement of the two methods was 89% (n = 467), among which, 12% (n = 62) were positive for both CA 15.3 and CA 27.29 (above the upper limit of the reference range), and 77% (n = 405) were negative for both assays (within the reference range). The disagreement was 11% (n = 56), which showed positive CA 27.29 and negative CA 15.3. The correlation coefficient between CA 15.3 and CA 27.29 is 0.9085 when both are positive and 0.6315 when both are negative. The bias is –238.9 (–84.7%) when both are positive and –8.5 (–52.4%) when both are negative. Paired t tests showed significant difference (P < .001) between the CA 15.3 and CA 27.29 in both positive samples and both negative samples. Conclusion In summary, there is a good correlation between CA 15.3 tested on Beckman Coulter UniCel® DxI 800 and CA 27.29 on ADVIA Centaur System. However, with the negative bias, they should not be used interchangeably.

Circulating tumor cells as a response monitor in stage IV non-small cell lung cancer

BackgroundMonitoring circulating tumor cells (CTC) has been shown to be prognostic in most solid malignancies. There is no CTC assay in clinical use for lung cancer therapy monitoring due to inconclusive clinical utility data. Limited data has been published outside of the standard CTC enumerations, regarding clinical significance of phenotypic heterogeneity of CTCs in late stage NSCLC and its ability to correlate with treatment outcomes.MethodsIn 81 patients with stage IV NSCLC, multiple timepoints for CTC analysis were collected after initiation of treatment across 139 lines of therapy using single cell high definition diagnostic pathology imaging of all nucleated cells from 362 peripheral blood samples as a liquid biopsy.ResultsWe analyzed the subset of 25 patients with complete time series data, totaling 117 blood samples, to determine the significance of HD-CTC kinetics during the initiation of treatment. These kinetics follow three distinct patterns: an increase in HD-CTCs with therapy (mean + 118.40 HD-CTCs/mL), unchanged HD-CTCs numbers (stable; mean 0.54 HD-CTCs/mL), and a decrease in HD-CTCs numbers (mean − 81.40 HD-CTCs/mL). Patients with an increasing CTC count during the first 3 months post initiation of new treatment had a better PFS and OS compared to the other groups. There was weak correlation between the absolute number of HD-CTCs at a single time point of therapy and patient outcomes (OS p value = 0.0754). In the whole cohort of 81 patients, HD-CTCs were detected in 51 (63%) patients at initiation of therapy with a median of 2.20 (range 0–509.20) and a mean of 26.21 HD-CTCs/mL (± 15.64).ConclusionsCTCs are identifiable in most patients with stage IV NSCLC. While absolute HD-CTC counts do not correlate with prognosis, the changes in CTC counts were predictive of survival in patients with metastatic lung cancer receiving chemotherapy. The level and dynamics of CTCs indicate very different biological and pharmacological phenomena at different stages of disease and timepoints of treatment, highlighting the complex role of CTCs in cancer research and clinical management.

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.

Extracellular Vesicles and Their Potential Use in Monitoring Cancer Progression and Therapy: The Contribution of Proteomics.

Extracellular Vesicles (EVs) are small membrane-enclosed particles released by cells and able to vehiculate information between them. The term EVs categorizes many and different vesicles based on their biogenesis and release pathway, such as exosomes (Exo), ectosomes, or shedding microvesicles (SMVs), apoptotic blebs (ABs), and other EVs subsets, generating a heterogeneous group of components able to redistribute their cargo into the entire organism. Moreover EVs are becoming increasingly important in monitoring cancer progression and therapy, since they are able to carry specific disease biomarkers such as Glypican-1, colon cancer-associated transcript 2, CD63, CD24, and many others. The importance of their biological role together with their heterogeneity prompted researchers to adopt and standardize purification methods able to isolate EVs for characterizing their cargo. In this way, mass spectrometry (MS)-based proteomics approaches are emerging as promising tool for the identification and quantification of EVs protein cargoes, but this technique resulted to be deeply influenced by the low quality of the isolation techniques. This review presents the state-of-the-art of EVs isolation, purification, and characterization for omics studies, with a particular focus to their potential use in monitoring cancer progression and therapy.

HE4 in therapy monitoring of advanced ovarian cancer: Comparison with CA 125

HE4 in therapy monitoring of advanced ovarian cancer: Comparison with CA 125

PSMA-ligand PET allows a more accurate therapeutic response evaluation of bone metastases in prostate cancer compared to computed tomography.

A patient with bone metastases of prostate cancer was referred for 68Ga-PSMA-11 PET/CT. Compared to a 68Ga-PSMA-11 PET/CT four months previously, the CT-component of the current PET/CT showed morphological progress in all lesions despite continuous therapy with docetaxel and denosumab. Contrarily, the PET-component showed a reduction of tracer-uptake, which correlated with PSA decrease between the two PET/CT-scans (16.2 ng/ml vs. 3.1 ng/ml). This case highlights 68Ga-PSMA-11 PET/CT as a promising tool for therapy monitoring of prostate cancer and could serve as a basis for a novel monitoring strategy. Volume progress shown by CT must not be classified as tumor progress, but as bone remodeling following effective therapy.

Liquid biopsy in tumor diagnostics : Applications, perspectives, and limitations of the "cancer liquidome"

The detection of tumor-specific genetic alterations in body fluids as an addition to or even replacement for established tissue-based tumor diagnostics is currently ahot topic in academic research and industry. Progress in methods for nucleic acid analyses together with promising results from clinical studies have raised great expectations for cancer screening, diagnosis, prognosis, and therapy monitoring by means of aminimally invasive blood draw. Individual focused assays have already been introduced into routine diagnostics and represent avaluable option in cases where no tissue samples are available. However, before the use of liquid biopsy outside of clinical studies is enforced and more complex markers (like tumor mutational burden) are analyzed, several practical challenges and principal problems have to be addressed. This review focusses on the detection of free-circulating nucleic acids in blood plasma and critically discusses established and future applications as well as challenges and limitations of this new method.

Use of IR Spectroscopy in Cancer Diagnosis. A Review

Results of studies of the use of FTIR spectroscopy in cancer diagnosis are reviewed. Materials studied include biological fluids, tissues, and model systems for studying experimental neoplasia. The methods for working with each system are described. A detailed description is given for the use of IR spectroscopy for cancer diagnosis and in the monitoring of cancer therapy for the evaluation of drug effectiveness and determination of the disease state. Statistical methods for processing the IR spectral data are presented. The main limitations to the use of IR spectroscopy for the diagnosis of oncological diseases and the potential for its introduction into clinical practice are described.

Probing the Cellular Size Distribution in Cell Samples Undergoing Cell Death

A polydisperse scattering model adapted for concentrated medium, namely the polydisperse structure factor model, was examined to explain the backscatter coefficients (BSCs) measured from packed cell samples undergoing cell death. Cell samples were scanned using high-frequency ultrasound in the 10–42 MHz bandwidth. A parameter estimation procedure was proposed to estimate the volume fraction and the relative impedance contrast that could explain the changes in BSC pattern by considering the actual change in cellular size distribution. Quantitative ultrasound parameters were estimated and related to the percentage of dead cells determined by flow cytometry. The standard deviation of scatterer size distribution extracted from the polydisperse structure factor model and the spectral intercept were found to be strongly correlated to the percentage of dead cells (r2 = 0.79 and r2 = 0.72, respectively). This study contributes to the understanding of ultrasonic scattering from cells undergoing cell death toward the monitoring of cancer therapy.

Hybrid magnetic and deformability based isolation of circulating tumor cells using microfluidics

Circulating tumor cell (CTC) and their isolation is one potential clinically useful target within a “liquid biopsy”. Detection and characterization of CTCs provide a non-invasive approach for monitoring cancer therapy. In the present research, a magnet- deformability hybrid integrated microfluidic chip was designed to enumerate CTCs. Arrays of narrow gaps (patterned by micro-ellipse microposts) were designed to capture CTCs bonded with magnetic immune beads. Eleven samples, including Gastric of Colorectal, non-small-cell lung cancer (NSCLC), Ureter cancer, and Breast cancer, were used to validate the microfluidic device clinically. Clinical test results show a high capture efficiency (over 90% at 3ml/h) and high viability (96%) at high flow rate. Furthermore, comparisons of device performance with and without magnet beads were also conducted. This hybrid chip design demonstrated excellent performance in the clinical enumeration of CTCs, which is promising in the future cancer therapy evaluation.

Breast Cancer Metabolomics Using NMR.

Continued progress is being made in understanding the breast cancer metabolism using analytical magnetic resonance (MR)-based methods like nuclear magnetic resonance (NMR) and in-vivo MR spectroscopy (MRS). Analyses using these methods have enhanced the knowledge of altered biochemical pathways associated with breast cancer progression, regression, and pathogenesis. Comprehensive metabolic profiling of biological samples like tissues, cell lines, fine needle aspirate, and biofluids such as sera and urine enables identification of new biomarkers and abnormalities in biochemical pathways. These methods are not only useful for diagnosis, therapy monitoring, disease progression, and staging of cancer but also for the identification of new therapeutic targets and designing new treatment strategies. Additionally, in-vivo MRS studies have established choline-containing compounds (tCho) as biomarkers of malignancy, which is useful for enhancing the diagnostic specificity of magnetic resonance imaging (MRI). Recent technological developments related to in-vivo MRS such as increased magnetic field strength, multichannel phased array breast coils, and absolute quantification of tCho have provided a better understanding of the tumor heterogeneity, metabolism, and pathogenesis. This chapter focuses on providing the experimental aspects of in-vitro, ex-vivo, and in-vivo MR spectroscopy methods used for metabolomics studies of breast cancer.

Circulating tumor cells (CTCs) for the noninvasive monitoring and personalization of non-small cell lung cancer (NSCLC) therapies.

Growing evidences for tumor heterogeneity confirm that single-tumor biopsies frequently fail to reveal the widespread mutagenic profile of tumor. Repeated biopsies are in most cases unfeasible, especially in advanced cancers. We describe here how circulating tumor cells (CTCs) isolated from minimally invasive blood sample might inform us about intratumor heterogeneity, tumor evolution and treatment resistance. We also discuss the advances of CTCs research, most notably in molecularly selected non-small cell lung cancer (NSCLC) patients, highlighting challenges and opportunities related to personalized therapy.

Fabrication of perforated polyethylene microfiltration membranes for circulating tumor cells separation by thermal nanoimprint method

Size-based filtration has been proved an efficient and quick approach to separate circulating tumor cells (CTCs) from other blood cells for cancer diagnosis and therapy monitoring. In this work, we proposed a simple, cost efficient and scalable approach to fabricate microporous polyethylene (PE) membranes for CTC filtration by thermal nanoimprint method. PE was selected as the material for the membrane due to its commercially available films with a thickness as thin as ~ 10 µm, which is matched well with the size of the micropores for CTCs and critical to a successful nanoimprint with a low and uniform residual layer. A thermal imprint process with a nickel mold was applied to fabricate periodic microporous PE membranes with a pitch of 20 µm and diameter of 10 µm. The perforated micropores were obtained by a short time O2 plasma-etching to remove the imprint residual layer. This PE membrane microfilter achieved 84% average capture efficiency for lung cancer cells spiked in blood samples.

Circulating Cell-Free DNA and Cancer Therapy Monitoring: Methods and Potential.

The monitoring of therapy during the treatment of cancer patients is currently assessed by the levels of circulating tumor cells or by PET/CT scans. Neither approach has the sensitivity or specificity to be very sure of the efficacy of the treatment. Moreover, PET/CT scans can be both comparatively expensive and produce low levels of radiation for the patient. The advent of the possibility of using circulating DNA released from the tumor permits (1) a possible early marker of the presence of the cancer, (2) an indication of the success of the primary treatment, (3) an indication of the early presence of possible metastasis, (4) a marker of the success of secondary subsequent treatment, (5) determining which patients can benefit from a particular treatment, and (6) offering a prognosis. These aspects will be discussed concerning the application of circulating tumor DNA analysis to the monitoring of cancer patients undergoing therapy.

Colorimetric Analysis of Carcinoembryonic Antigen Using Highly Catalytic Gold Nanoparticles-Decorated MoS2 Nanocomposites.

Sensitive detection of carcinoembryonic antigen (CEA) is very important for early detection and cancer therapy monitoring. We utilized the highly catalytic activity of MoS2-gold nanoparticles (MoS2-AuNPs) nanocomposites to construct a colorimetric immunoassay for CEA detection, which could catalyze sodium borohydride (NaBH4) to reduce 4-nitrophenol (4-NP) and make the yellow solution colorless. MoS2-AuNPs nanocomposites could efficiently load antibodies (Ab1 and Ab2) because of their good biocompatibility and high adsorption ability, forming Ab1-MoS2-AuNPs nanoprobes for capturing CEA and Ab2-MoS2-AuNPs nanoprobes for catalytic reaction. It should be pointed out that Ab2-MoS2-AuNPs nanoprobes retained high catalytic activity because no blocking agent was used to block the catalytic sites. After the formation of the classical "sandwich" structure, parts of Ab2-MoS2-AuNPs nanoprobes remained in the supernatant after centrifugation, making the catalytic reaction occur and obtaining a colorless solution. As expected, a linear function between absorbance peak intensity and the logarithm of 5 pg/mL-10 ng/mL CEA was observed, and the detection limit was obtained as 0.5 pg/mL. This colorimetric immunosensor showed acceptable selectivity and repeatability for CEA detection, which could be successfully employed to detect CEA in serum.

Oxygen-Enhanced and Dynamic Contrast-Enhanced Optoacoustic Tomography Provide Surrogate Biomarkers of Tumor Vascular Function, Hypoxia, and Necrosis.

Measuring the functional status of tumor vasculature, including blood flow fluctuations and changes in oxygenation, is important in cancer staging and therapy monitoring. Current clinically approved imaging modalities suffer long procedure times and limited spatiotemporal resolution. Optoacoustic tomography (OT) is an emerging clinical imaging modality that may overcome these challenges. By acquiring data at multiple wavelengths, OT can interrogate hemoglobin concentration and oxygenation directly and resolve contributions from injected contrast agents. In this study, we tested whether two dynamic OT techniques, oxygen-enhanced (OE) and dynamic contrast-enhanced (DCE)-OT, could provide surrogate biomarkers of tumor vascular function, hypoxia, and necrosis. We found that vascular maturity led to changes in vascular function that affected tumor perfusion, modulating the DCE-OT signal. Perfusion in turn regulated oxygen availability, driving the OE-OT signal. In particular, we demonstrate for the first time a strong per-tumor and spatial correlation between imaging biomarkers derived from these in vivo techniques and tumor hypoxia quantified ex vivo Our findings indicate that OT may offer a significant advantage for localized imaging of tumor response to vascular-targeted therapies when compared with existing clinical DCE methods.Significance: Imaging biomarkers derived from optoacoustic tomography can be used as surrogate measures of tumor perfusion and hypoxia, potentially yielding rapid, multiparametric, and noninvasive cancer staging and therapeutic response monitoring in the clinic.Graphical Abstract: http://cancerres.aacrjournals.org/content/canres/78/20/5980/F1.large.jpg Cancer Res; 78(20); 5980-91. ©2018 AACR.