Detect Cancer Metastases Research Articles

Abstract B005: A robust NSCLC biomarker: miR-7-5p: in-silico validation and potential SPR- based probe for detection

Abstract The abundance of micro RNAs as a specific biomarker for accurately detecting cancer metastasis has arisen recently. The expression levels of miRNA are analyzed to get insights into cancer tissue detection and subtypes. Similar to other cancer types, the miRNA shows high levels of target mRNA dysregulation in association with non-small cell lung carcinoma (NSCLC). Among many promising cancer biomarkers for NSCLC, miR-7-5p has shown significant downregulation in the NSCLC tissues and targets proto-oncogenes like PAK2 and NOVA2. The expression levels of different proto-oncogenes targeting the miR-7-5p in NSCLC show that the EGFR- mutated NSCLC has an experimental validation. And, the target validation of the miR-7-5p could be analyzed using SPR (Surface plasmon resonance) based sensors at a single nanoparticle level such as Au nanocube due to its high specificity and accountability. Despite being an accountable tool for cancer diagnosis, miRNA-based biomarkers sometimes cause poor diagnostic specificity and reproducibility due to their heterogenicity and immunogenicity in cancer detection. To overcome these shortcomings, the biomarkers need to be validated according to recent clinical studies. Citation Format: Chandrajeet Dhara, Anindita Dhara, Saumyatika Gantayat. A robust NSCLC biomarker: miR-7-5p: in-silico validation and potential SPR- based probe for detection [abstract]. In: Proceedings of the AACR Special Conference in Cancer Research: RNAs as Drivers, Targets, and Therapeutics in Cancer; 2024 Nov 14-17; Bellevue, Washington. Philadelphia (PA): AACR; Mol Cancer Ther 2024;23(11_Suppl):Abstract nr B005.

Analytical solution of a microrobot-blood vessel interaction model

This study develops a dynamics model of a microrobot vibrating in a blood vessel aiming to detect potential cancer metastasis. We derive an analytical solution for microrobot’s motion, considering interactions with the vessel walls modelled by a linear spring-dashpot and a constant damping value for blood viscosity. The model facilitates instantaneous state transitions of the microrobot, such as contact with the vessel wall and free motion within the fluid. Amplitudes and phase angles from the transient solutions of dynamics model of the microrobot are solved at arbitrary moments, providing insights into its transient dynamics. The analytical solution of the proposed system is validated by experimental data, serving as a benchmark to examine the influence of pertinent parameters on microrobot’s dynamic response. It is found that the contact force transmitted to the vessel wall, assessed by system’s transmissibility function dependent on damping and frequency ratios, decreases with increasing damping ratio and intensifies when the frequency ratio is below 2. At the frequency ratio is equal to 1, resonance phenomenon is dominated by the magnification factor linked to the damping ratio, increasing the amplitude of resonance as damping decreases. Finally, different sets of system parameters, including excitation frequency and magnitude, fluid damping, vessel wall’s stiffness and damping, reveal multi-periodic motions and fake collision of the microrobot with the vessel wall. Simulation results imply that these phenomena are minimally affected by vessel wall’s stiffness but are significantly influenced by other parameters, such as fluid damping coefficient and damping coefficient of the blood vessel wall. This research provides a robust theoretical foundation for developing control strategies for microrobots aimed at detecting cancer metastasis.

Bifunctional electrochemical biosensor based on PB-MXene films for the real-time analysis and detection of living cancer cells

Circulating tumor cells (CTCs) are important prognostic markers for cancer diagnosis and metastasis, and their detection is an important means to detect cancer metastasis. Herein, we construct a novel bifunctional electrochemical biosensor based on the PB-MXene composite films. A simple electrostatic self-assembly approach was employed to prepare a film composed of PB nanocubes on the MXene substrates. Given that the PB is an artificial peroxidase for H2O2 sensing, the PB-MXene films can realize the real-time monitoring of H2O2 secretion from living CTCs. Besides, the anti-CEA attached biosensors can be utilized to quantify the corresponding CTCs. The synergic effects of the MXene with a large specific area and PB with enzyme-free catalysis for H2O2 resulted in PB-MXene films exhibiting high electrocatalytic and low cytotoxicity for both H2O2 sensing and living CTCs capturing. As a result, the biosensor shows a low detection limit of 0.57 μM towards H2O2 with a wide linear range (1 μM to 500 μM), as well as an excellent sensing performance for CTCs (an extremely low detection limit of 9 cells/mL in a wide linear range of 1.3 ×101 to 1.3 ×106 cells/mL). Moreover, the prepared biosensor showed satisfactory stability and anti-interference ability for potential applications in clinical cancer diagnosis and tumor metastasis.

Automatic detection of cancer metastasis in lymph node using deep learning

Lymph node metastases are one of the most indicator of some cancer types such as breast, colon and prostate. Breast cancer mostly spreads to lymph nodes in the armpit and it is one of the most common causes of death in women worldwide. Pathologists need more attention and time to diagnose metastasis in digitized lymph node images and also this process tends to be misinterpreted. In this paper, a promising deep learning based approach was presented to detect cancer metastasis in lymph node images with higher accuracy. The proposed deep learning framework uses ResNet architectures, transfer learning and 1cycle policy which is the method of finding the optimal learning rate. The validity of the proposed approach was evaluated on PCAM dataset consisted of 220,025 lymph node images. The experiments demonstrated that the presented method outperformed most of the current studies, achieving accuracy of 98.60% on the PCAM dataset by using ResNet models and fine-tuning techniques effectively. The burden of diagnosis procedure of metastasis can be lessened with the proposed deep learning framework in the clinical applications.

Biocompatible nanomicelles for sensitive detection and photodynamic therapy of early-stage cancer.

The lack of sensitive detection techniques and agents for early-stage tumors, which are characterized by small size, juvenile blood vessels and scarce secreted markers, has hampered timely cancer therapy and human well-being. Herein, the natural product pyropheophorbide-a (PPa) and FDA-approved Pluronic F127 are organized to develop F127-PPa nanomicelles with favorable size, red-shifted fluorescence and decent biocompatibility. After intravenous (i.v.) injection, the F127-PPa nanomicelles could not only accurately identify early-stage xenografted tumors, but also sensitively detect cancer metastasis in lungs through near-infrared (NIR) fluorescence imaging. The fluorescence signals are consistent with radionuclide imaging, photoacoustic (PA) imaging and bioluminescence imaging of tumors, consolidating the reliability of using F127-PPa nanomicelles for sensitive cancer diagnosis in a non-invasive and low-cost manner. Moreover, the fluorescence intensity of small tumors is linearly correlated with the tumoral mass ranging from 10 to 120 mg with a fluorescence coefficient of 4.5 × 107 mg-1. Under the guidance of multimodal imaging, the tumors could be thoroughly eradicated by F127-PPa under laser irradiation due to efficient reactive oxygen species (ROS) generation. These findings may provide clinically translatable agents and strategies for sensitive diagnosis of early-stage tumors and timely cancer therapy.

Detecting cancer metastasis and accompanying protein biomarkers at single cell levels using a 3D-printed microfluidic immunoarray

A low-cost microfluidic microarray capable of lysing cells and quantifying proteins released after lysis was designed and 3D-printed. The array lyses cells on-chip in lysis buffer augmented with a 2s pulse of a sonic cell disruptor. Detection of desmoglein 3 (DSG3), a metastatic biomarker for head and neck squamous cell carcinoma (HNSCC), along with two accompanying HNSCC biomarkers from a single cell lysate of oral cancer cell cultures was demonstrated. A lysis chamber and reagent compartments deliver sample and reagents into detection chambers decorated with capture antibodies immobilized onto inner walls coated with a highly swollen 3D chitosan hydrogel film. Sandwich immunoassays are achieved when captured analytes labeled with biotinylated secondary antibodies, which then capture streptavidin-poly [horse radish peroxidase] (Poly-HRP). Subsequent delivery of super-bright femto-luminol with H2O2 generates chemiluminescence captured with a CCD camera. DSG3 is membrane-bound protein in HNSCC cells of invaded lymph nodes, vascular endothelial growth factor-A (VEGF-A), vascular endothelial growth factor-C (VEGF-C) were positive controls overexpressed into the HNSCC culture medium. Beta-tubulin (β-Tub) was used as a loading control to estimate the number of cells in analyzed samples. Limits of detection (LOD) were 0.10 fg/mL for DSG3, and 0.20 fg/mL for VEGF-A, VEGF-C and β-Tub. Three orders of magnitude semilogarithmic dynamic ranges were achieved. VEGF-A showed high in-cell expression, but VEGF-C had low levels inside cells. The very low LODs enabled quantifying these proteins released from single cells. Strong correlation between results from on-chip cell lysis, conventional off-line lysis and ELISA confirmed accuracy.

The biomarker features of miR-145-3p determined via meta-analysis validated by qRT-PCR in metastatic cancer cell lines

MicroRNAs (miRNAs) play important roles in the cancer biology such as proliferation, differentiation, and apoptosis. The pivotal roles that miRNA expression plays, make them ideal candidates for detection of cancer progression as well as cancer metastasis. Especially for breast, lung and prostate cancer which are originated from soft tissues and prone to metastasis. Thus, the aim of this study is to evaluate the expression level of miR-145-3p which is a shared potential biomarker identified by meta-analysis of breast, prostate and lung cancer data sets.Six different data sets representative of three different cancer types were analyzed. These data sets are pooled together to have a master metamiRNA list while getting rid of the platform differentiations between them. As a result, 24 common differentially expressed miRNAs are determined in which miR-145-3p has the topmost rank. To mimic in vivo cancer microenvironment, hypoxia and serum deprivation were used to induce metastasis in breast (MCF-7, MDA-MB-231, MDA-MB-453), prostate (PC3, LNCaP, DU145), lung (A549, NCIH82,) cancer cell lines and noncancerous cell lines of the coresponding tissues (MCF10A, RWPE-1, MRC-5).miR-145-3p expression levels were determined by qRT-PCR. It has been shown that it is down regulated by the induction of metastasis in cancer cell lines while it is up regulated in normal cell lines to suppress the tumor formation. As a conclusion, as representing the same results in three different cancer cell types, miR-145-3p will be a promising biomarker to follow up its expression to detect cancer metastasis.

Indocyanine-green-loaded microbubbles for localization of sentinel lymph node using near-infrared fluorescence/ultrasound imaging: a feasibility study

Current strategies for sentinel lymph node (SLN) biopsy to detect cancer metastasis have some limitations such as the associated radiation exposure and high false-negative rates due to dye particles through the true SLNs to contiguous LNs.

MicroRNAs used as novel biomarkers for detecting cancer metastasis.

The low survival rates of cancers are primarily due to late diagnosis and metastasis. Discriminating the metastasis is a crucial factor for prognosis and improving the survival rate of cancer patients. MicroRNAs (miRNAs) can regulate the expression of hundreds of downstream genes, which has a broad effect on the regulation of the whole cell cycle. Accumulating studies have found that the aberrant expression of miRNAs is associated with cancer genesis. The aim of this study is to evaluate the diagnostic value of miRNAs in detecting cancer metastasis. Medline, PubMed, Embase, and CNKI were searched for relevant articles. Sensitivity, specificity, positive and negative likelihood ratio (PLR, NLR) and diagnostic odds ratio (DOR), the summary receiver operator characteristic (SROC) curve and the calculated AUC (area under the SROC curve) were applied to explore the diagnostic accuracy of miRNAs in metastasis. Seven hundred seventy-one metastatic cancer patients and 552 non-metastatic cancer controls from 14 articles were involved in our meta-analysis. A sensitivity of 0.75 (95% confidence interval (CI), 0.72-0.79) and a specificity of 0.80 (95% CI, 0.76-0.84) were observed from metastatic patients and non-metastatic controls in the combined analysis. And the AUC was 0.83 (95% CI, 0.79-0.86). In addition, results from subgroup analyses suggested that a higher diagnostic value for metastasis was acquired in tissue sample other than blood sample (sensitivity, 0.82 versus 0.73; specificity, 0.84 versus 0.79; PLR, 5.0 versus 3.5; NLR, 0.22 versus 0.34; DOR, 23 versus 10; AUC, 0.88 versus 0.80). In summary, this meta-analysis proved the relatively high diagnostic value of miRNA in metastasis, which might be applied as a novel screening tool to detect metastasis along with other biomarkers. We also illustrated that tissue-based miRNAs may have a better diagnostic accuracy than blood-based miRNAs.

Near IR Heptamethine Cyanine Dye–Mediated Cancer Imaging

Near-IR fluorescence imaging has great potential for noninvasive in vivo imaging of tumors. In this study, we show the preferential uptake and retention of two hepatamethine cyanine dyes, IR-783 and MHI-148, in tumor cells and tissues. IR-783 and MHI-148 were investigated for their ability to accumulate in human cancer cells, tumor xenografts, and spontaneous mouse tumors in transgenic animals. Time- and concentration-dependent dye uptake and retention in normal and cancer cells and tissues were compared, and subcellular localization of the dyes and mechanisms of the dye uptake and retention in tumor cells were evaluated using organelle-specific tracking dyes and bromosulfophthalein, a competitive inhibitor of organic anion transporting peptides. These dyes were used to detect human cancer metastases in a mouse model and differentiate cancer cells from normal cells in blood. These near-IR hepatamethine cyanine dyes were retained in cancer cells but not normal cells, in tumor xenografts, and in spontaneous tumors in transgenic mice. They can be used to detect cancer metastasis and cancer cells in blood with a high degree of sensitivity. The dyes were found to concentrate in the mitochondria and lysosomes of cancer cells, probably through organic anion transporting peptides, because the dye uptake and retention in cancer cells can be blocked completely by bromosulfophthalein. These dyes, when injected to mice, did not cause systemic toxicity. These two heptamethine cyanine dyes are promising imaging agents for human cancers and can be further exploited to improve cancer detection, prognosis, and treatment.

Dye- and gamma probe-guided sentinel lymph node biopsy in breast cancer patients: using patent blue dye and technetium-99m-labeled human serum albumin

Sentinel lymph node (SLN) biopsy is a promising method for the diagnosis of the axillary nodal status. We examined the availability of the SLN biopsy using two mapping procedures: the dye- and gamma probe-guided method, and preoperative lymphoscintigraphy by gamma camera imaging. We enrolled 48 patients with breast cancer. Technetium-99m-labeled human serum albumin was injected into the subdermal tissue above the primary tumor or biopsy cavity, and preoperative gamma camera imaging was performed. After induction of general anesthesia, patent blue dye was injected into the peritumoral area prior to the surgical procedure. A handheld gamma-detection probe was used to assist in SLN detection. Careful dissection was performed to identify blue-stained afferent lymphatic vessels and nodes. An SLN was defined as any blue and/or radioactive node, and was excised. After SLN biopsy, axillary lymph node dissection of level I, II, and III was completed, in order to confirm the diagnostic ability of the SLN biopsy. Intraoperative SLN identification of axillary lesions was successful in 43 of 48 patients (90%). The dye- and gamma probe-guided method was successful in 25 patients (52%), the dye-guided method alone succeeded in 11 patients (23%), and the gamma probe-guided method alone succeeded in 7 patients (15%). Preoperative lymphoscintigraphy revealed axillary focal accumulations in 29 of 48 patients (60%). All patients who underwent successful preoperative SLN identification by lymphoscintigraphy had successful intraoperative SLN identification. A diagnostic accuracy of 95%, a sensitivity of 89%, and a specificity of 100% were achieved in the diagnosis of axillary metastasis. Internal mammary SLNs were identified in four patients intraoperatively, but we could not detect cancer metastasis in the internal mammary SLNs. The dye-guided and gamma probe-guided methods were complementary. Preoperative lymphoscintigraphy was useful to predict intraoperative SLN identification. Further study is necessary to assess the role of SLN biopsy of the internal mammary lymph nodes.

Tracing the in VivoDistribution and Dynamics of Cancer Cells in Mice by Luminescence of Aequorin- Expressing Transformants

In order to trace the in vivodynamics of cancer cells, we introduced jellyfish aequorin cDNA into metastatic (A-11) and nonmetastatic (3LL) Lewis lung cancer cell lines. The transformants ( e.g.,A-11-4 and 3LL-24) remained highly tumorigenic when they were s.c.injected into C57BL/6 mice and the A-11-4 clone produced metastatic tumors in the lung. Luminescence produced by the expressed aequorin was detectable in these transformants in culture as well as in cells of primary or secondary tumors. The limit of detection of the aequorin assay was ≈90 cells, whereas ≈5 ×10 4cells could be macroscopically seen as metastatic foci in the lung. Thus, the aequorin assay is ≈500-fold more sensitive than macroscopic examination in detecting cancer metastasis.