Dataset Information

Multiplexed molecular imaging with surface enhanced resonance Raman scattering nanoprobes reveals immunotherapy response in mice via multichannel image segmentation.

ABSTRACT: Visualizing the presence and distribution of multiple specific molecular markers within a tumor can reveal the composition of its microenvironment, inform diagnosis, stratify patients, and guide treatment. Raman imaging with multiple molecularly-targeted surface enhanced Raman scattering (SERS) nanoprobes could help investigate emerging cancer treatments preclinically or enable personalized treatment assessment. Here, we report a comprehensive strategy for multiplexed imaging using SERS nanoprobes and machine learning (ML) to monitor the early effects of immune checkpoint blockade (ICB) in tumor-bearing mice. We used antibody-functionalized SERS nanoprobes to visualize 7 + 1 immunotherapy-related targets simultaneously. The multiplexed images were spectrally resolved and then spatially segmented into superpixels based on the unmixed signals. The superpixels were used to train ML models, leading to the successful classification of mice into treated and untreated groups, and identifying tumor regions with variable responses to treatment. This method may help predict treatment efficacy in tumors and identify areas of tumor variability and therapy resistance.

SUBMITTER: Andreou C

PROVIDER: S-EPMC10360075 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature

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Publications

Multiplexed molecular imaging with surface enhanced resonance Raman scattering nanoprobes reveals immunotherapy response in mice <i>via</i> multichannel image segmentation.

Andreou Chrysafis C Plakas Konstantinos K Berisha Naxhije N Gigoux Mathieu M Rosch Lauren E LE Mirsafavi Rustin R Oseledchyk Anton A Pal Suchetan S Zamarin Dmitriy D Merghoub Taha T Detty Michael R MR Kircher Moritz F MF

Nanoscale horizons 20221121 12

Visualizing the presence and distribution of multiple specific molecular markers within a tumor can reveal the composition of its microenvironment, inform diagnosis, stratify patients, and guide treatment. Raman imaging with multiple molecularly-targeted surface enhanced Raman scattering (SERS) nanoprobes could help investigate emerging cancer treatments preclinically or enable personalized treatment assessment. Here, we report a comprehensive strategy for multiplexed imaging using SERS nanoprob ...[more]

PMID: 36285605

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Project description:Azo compounds such as the Sudan dyes I-IV are frequently used illegally as colorants and added to a wide range of foods. These compounds have been linked to a number of food safety hazards. Several methods have been proposed to detect food contamination by azo compounds and most of these are laboratory based; however, the development of reliable and portable methods for the detection and quantification of food contaminated by these chemicals in low concentration is still needed due to their potentially carcinogenic properties. In this study, we investigated the ability of surface enhanced Raman scattering (SERS) combined with chemometrics to quantify Sudan I-IV dyes. SERS spectra were acquired using a portable Raman device and gold nanoparticles were employed as the SERS substrate. As these dyes are hydrophobic, they were first dissolved in water: acetonitrile (1:10, v/v) as single Sudan dyes (I-IV) at varying concentrations. SERS was performed at 785 nm and the spectra were analyzed by using partial least squares regression (PLS-R) with double cross-validations. The coefficient of determination (Q2) were 0.9286, 0.9206, 0.8676 and 0.9705 for Sudan I to IV, respectively; the corresponding limits of detection (LOD) for these dyes were estimated to be 6.27 × 10-6, 5.35 × 10-5, 9.40 × 10-6 and 1.84 × 10-6 M. Next, quadruplex mixtures were made containing all four Sudan dyes. As the number of possible combinations needed to cover the full concentration range at 5% intervals would have meant collecting SERS spectra from 194,481 samples (214 combinations) we used a sustainable solution based on Latin hypercubic sampling and reduced the number of mixtures to be analyzed to just 90. After collecting SERS spectra from these mixture PLS-R models with bootstrapping validations were employed. The results were slightly worse in which the Q2 for Sudan I to IV were 0.8593, 0.7255, 0.5207 and 0.5940 when PLS1 models (i.e., one model for one dye) was employed and they changed to 0.8329, 0.7288, 0.5032 and 0.5459 when PLS2 models were employed (i.e., four dyes were modelled simultaneously). These results showed the potential of SERS to be used as a high-throughput, low-cost, and reliable methods for detecting and quantifying multiple Sudan dyes in low concentration from illegally adulterated samples.

Project description:Circulating tumor cells (CTCs) have substantial clinical implications in cancer diagnosis and monitoring. Although significant progress has been made in developing technologies for CTC detection and counting, the ability to quantitatively detect multiple surface protein markers on individual tumor cells remains very limited. In this work, we report a multiplexed method that uses magnetic multicolor surface-enhanced Raman scattering (SERS) nanotags in conjunction with a chip-based immunomagnetic separation to quantitatively and simultaneously detect four surface protein markers on individual tumor cells in whole blood. Four-color SERS nanotags were prepared using magnetic-optical iron oxide-gold core-shell nanoparticles with different Raman reporters to recognize four different cancer markers with respective antibodies. A microfluidic device was fabricated to magnetically capture the nanoparticle-bound tumor cells and to perform online negative staining and single-cell optical detection. The level of each targeted protein was obtained by signal deconvolution of the mixed SERS signals from individual tumor cells using the classic least squares regression method. The method was tested with spiked tumor cells in human whole blood with three different breast cancer cell lines and compared with the results of purified cancer cells suspended in a phosphate buffer solution. The method, with either spiked cancer cells in blood or purified cancer cells, showed a strong correlation with purified cancer cells by enzyme-linked immunosorbent assay, suggesting the potential of our method for the reliable detection of multiple surface markers on CTCs. Combining immunomagnetic enrichment with high specificity, multiplexed targeting for the capture of CTC subpopulations, multicolor SERS detection with high sensitivity and specificity, microfluidics for handling rare cells and magnetic-plasmonic nanoparticles for dual enrichment and detection, our method provides an integrated, yet a simple and an efficient platform that has the potential to more sensitively detect and monitor cancer metastasis.

Dataset Information

Multiplexed molecular imaging with surface enhanced resonance Raman scattering nanoprobes reveals immunotherapy response in mice via multichannel image segmentation.

Publications

Multiplexed molecular imaging with surface enhanced resonance Raman scattering nanoprobes reveals immunotherapy response in mice <i>via</i> multichannel image segmentation.

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