Acid specific dark quencher QC1 pHLIP for multi-spectral optoacoustic diagnoses of breast cancer

Breast cancer is the most common type of malignant growth in women. Early detection of breast cancer, as well as the identification of possible metastatic spread poses a significant challenge because of the structural and genetic heterogeneity that occurs during the progression of the disease. Currently, mammographies, biopsies and MRI scans are the standard of care techniques used for breast cancer diagnosis, all of which have their individual shortfalls, especially when it comes to discriminating tumors and benign growths. With this in mind, we have developed a non-invasive optoacoustic imaging strategy that targets the acidic environment of breast cancer. A pH low insertion peptide (pHLIP) was conjugated to the dark quencher QC1, yielding a non-fluorescent sonophore with high extinction coefficient in the near infrared that increases signal as a function of increasing amounts of membrane insertion. In an orthotopic murine breast cancer model, pHLIP-targeted optoacoustic imaging allowed us to differentiate between healthy and breast cancer tissues with high signal/noise ratios. In vivo, the sonophore QC1-pHLIP could detect malignancies at higher contrast than its fluorescent analog ICG-pHLIP, which was developed for fluorescence-guided surgical applications. PHLIP-type optoacoustic imaging agents in clinical settings are attractive due to their ability to target breast cancer and a wide variety of other malignant growths for diagnostic purposes. Intuitively, these agents could also be used for visualization during surgery

Advancing Breath‐Based Diagnostics: 3D Mesh SERS Sensor Via Dielectrophoretic Alignment of Solution‐Processed Au Nanoparticle‐Decorated TiO2 Nanowires

Abstract Surface enhanced Raman spectroscopy (SERS) is becoming an attractive analytical technique for the next generation of breath diagnostics. However, current SERS substrates present challenges related to fabrication cost, complexity, signal uniformity, and reproducibility. Here, a low‐cost, label‐free SERS sensor based on fully solution‐processed decoration of TiO2 nanowires is demonstrated (NW) with plasmonic Au nanoparticles (NP) followed by the dielectrophoretic self‐assembly into a 3D mesh with high signal to noise ratio. The sensor performance is tested using 4‐aminothiophenol (4‐ATP) as a model analyte in gas phase, at concentrations down to 10 ppbv, and in solution, with limit of detection ≈2.4 pM. Finally, to explore the sensor capability for breath‐based diagnostics, a proof‐of‐concept experiment is performed with exhaled breath condensates (EBCs). The possibility to discriminate EBCs of individuals with upper respiratory tract infection (URTI) from healthy ones is demonstrated. Multiple SERS spectra (n≈50) from each sample are analyzed using orthogonal partial least squares discriminant analysis (OPLS‐DA), which identifies spectral features representative of URTI in up to 80% of the infection‐related spectra. These results demonstrate the applicability and potential of 1D nanomaterials together with state‐of‐the‐art solution‐processed techniques for the development of low‐cost and compact SERS breath‐based diagnostic platforms for clinical point‐of‐care applications

Folate-Targeted Surface-Enhanced Resonance Raman Scattering Nanoprobe Ratiometry for Detection of Microscopic Ovarian Cancer

Ovarian cancer has a unique pattern of metastatic spread, in that it initially spreads locally within the peritoneal cavity. This is in contrast to most other cancer types, which metastasize early on <i>via</i> the bloodstream to distant sites. This unique behavior opens up an opportunity for local application of both therapeutic and imaging agents. Upon initial diagnosis, 75% of patients already present with diffuse peritoneal spread involving abdominal organs. Complete resection of all tumor implants has been shown to be a major factor for improved survival. Unfortunately, it is currently not possible for surgeons to visualize microscopic implants, impeding their removal and leading to tumor recurrences and poor outcomes in most patients. Thus, there is a great need for new intraoperative imaging techniques that can overcome this hurdle. We devised a method that employs folate receptor (FR)-targeted surface-enhanced resonance Raman scattering (SERRS) nanoparticles (NPs), as folate receptors are typically overexpressed in ovarian cancer. We report a robust ratiometric imaging approach using anti-FR-SERRS-NPs (αFR-NPs) and nontargeted SERRS-NPs (nt-NPs) multiplexing. We term this method “topically applied surface-enhanced resonance Raman ratiometric spectroscopy” (TAS3RS (“tasers”) for short). TAS3RS successfully enabled the detection of tumor lesions in a murine model of human ovarian adenocarcinoma regardless of their size or localization. Tumors as small as 370 μm were detected, as confirmed by bioluminescence imaging and histological staining. TAS3RS holds promise for intraoperative detection of microscopic residual tumors and could reduce recurrence rates in ovarian cancer and other diseases with peritoneal spread

Probing Aberrantly Glycosylated Mucin 1 in Breast Cancer Extracellular Vesicles

Aberrantly glycosylated mucin 1 is a critical prognostic biomarker in breast epithelial cancers. Hypoglycosylated mucin 1 coats the surface of the cancer cells, where O-glycans are predominantly linked via an N-acetylgalactosamine moiety (GalNAc). Cancer cell-derived extracellular vesicles (EVs) carry biomarkers from parent cancer cells to the recipient cells and, therefore, could potentially be leveraged for diagnostics and noninvasive disease monitoring. We devised a label-free approach for identifying glycoprotein mucin 1 overexpression on breast cancer EVs. While exploring a plethora of biochemical (enzyme-linked immunosorbent assay, flow cytometry, and SDS-PAGE) and label-free biophysical techniques (circular dichroism and infrared spectroscopy (IR)) along with multivariate analysis, we discovered that mucin 1 is significantly overexpressed in breast cancer EVs and aberrant glycosylation in mucin 1 could be critically addressed using IR and multivariate analysis targeting the GalNAc sugar. This approach emerges as a convenient and comprehensive method of distinguishing cancer EVs from normal samples and holds potential for nonintrusive breast cancer liquid biopsy screening

Aggregation Kinetics of SERS-Active Nanoparticles in Thermally Stirred Sessile Droplets

The aggregation kinetics of silver nanoparticles in sessile droplets were investigated both experimentally and through numerical simulations as a function of temperature gradient and evaporation rate, in order to determine the hydrodynamic and aggregation parameters that lead to optimal surface-enhanced Raman spectroscopic (SERS) detection. Thermal gradients promote effective stirring within the droplet. The aggregation reaction ceases when the solvent evaporates forming a circular stain consisting of a high concentration of silver nanoparticle aggregates, which can be interrogated by SERS leading to analyte detection and identification. We introduce the aggregation parameter, Γ_a  τ_evap/τ_a, which is the ratio of the evaporation to the aggregation time scales. For a well-stirred droplet, the optimal condition for SERS detection was found to be Γ_a,opt = <i>kc</i>_NPτ_evap ≈ 0.3, which is a product of the dimerization rate constant (<i>k</i>), the concentration of nanoparticles (<i>c</i>_NP), and the droplet evaporation time (τ_evap). Near maximal signal (over 50% of maximum value) is observed over a wide range of aggregation parameters 0.05 < Γ_a < 1.25, which also defines the time window during which trace analytes can be easily measured. The results of the simulation were in very good agreement with experimentally acquired SERS spectra using gas-phase 1,4-benzenedithiol as a model analyte

Colon Cancer: From Epidemiology to Prevention

Colorectal cancer (CRC) is one of the most prevalent cancers affecting humans, with a complex genetic and environmental aetiology. Unlike cancers with known environmental, heritable, or sex-linked causes, sporadic CRC is hard to foresee and has no molecular biomarkers of risk in clinical use. One in twenty CRC cases presents with an established heritable component. The remaining cases are sporadic and associated with partially obscure genetic, epigenetic, regenerative, microbiological, dietary, and lifestyle factors. To tackle this complexity, we should improve the practice of colonoscopy, which is recommended uniformly beyond a certain age, to include an assessment of biomarkers indicative of individual CRC risk. Ideally, such biomarkers will be causal to the disease and potentially modifiable upon dietary or therapeutic interventions. Multi-omics analysis, including transcriptional, epigenetic as well as metagenomic, and metabolomic profiles, are urgently required to provide data for risk analyses. The aim of this article is to provide a perspective on the multifactorial derailment of homeostasis leading to the initiation of CRC, which may be explored via multi-omics and Gut-on-Chip analysis to identify much-needed predictive biomarkers