Micro-NMR for Rapid Molecular Analysis of Human Tumor Samples

Abstract

Although tumor cells obtained from human patients by image-guided intervention are a valuable source for diagnosing cancer, conventional means of analysis are limited. Here, we report the development of a quantitative micro-NMR (nuclear magnetic resonance) system for rapid, multiplexed analysis of human tumors. We implemented the technology in a clinical setting to analyze cells obtained by fine-needle aspirates from suspected lesions in 50 patients and validated the results in an independent cohort of another 20 patients. Single fine-needle aspirates yielded sufficient numbers of cells to enable quantification of multiple protein markers in all patients within 60 min. Moreover, using a four-protein signature, we report a 96% accuracy for establishing a cancer diagnosis, surpassing conventional clinical analyses by immunohistochemistry. Our results also show that protein expression patterns decay with time, underscoring the need for rapid sampling and diagnosis close to the patient bedside. We also observed a surprising degree of heterogeneity in protein expression both across the different patient samples and even within the same tumor, which has important implications for molecular diagnostics and therapeutic drug targeting. Our quantitative point-of-care micro-NMR technique shows potential for cancer diagnosis in the clinic.

Superparamagnetic Iron Oxide Nanoparticles in Biomedicine: Applications and Developments in Diagnostics and Therapy

“…with this method. In a clinical study on 70 test subjects with abdominal carcinomas, Haun et al [226] were able to show that this still experimental-preclinical method can detect tumor cells with an accuracy of 96 % using a 4-protein signature (MUC-1 +EGFR+HER2 +EpCAM), that protein signatures of tumor cells depend on the extraction site within the tumors (tumor heterogeneity), and the signature can change from the time of sampling to cytological analysis making it necessary to perform real-time analysis. In clinical application, this method could make it possible to perform real-time analyses of tumor or blood cells directly ex vivo after sampling so that therapeutic treatment strategies can be introduced more quickly and can be better tailored to the in vivo situation.…”

“…Further applications of magnetic nanoparticles (MNP) with a high potential for further clinical diagnostics describe the in vitro use of targeted MNP or SPIO in quantitative diagnostic magnetic resonance (DMR) for the highly sensitive detection of biomarkers [223] (DNA, mRNA, proteins, small molecules, enzymes, medications), pathogens (viruses [224], bacteria [225]), or cells (tumor cells [226]), circulating immune/tumor cells [227]) under modulation of the T2 relaxation of biological samples. Using miniaturized, chip-based DMR detector systems, it was able to be shown that highly sensitive, multiple DMR measurements can be performed using sample volumes of only several microliters (< 10 μl) [223].…”

Molecular imaging for personalized cancer care

“…A well-known example of this approach is the use of HER-2 testing of breast cancer tissue before the administration of trastuzumab, which targets the HER2 protein in breast cancer (Hricak, 2011). Revolutionary advances in the technology for in vitro diagnostics include nuclear MR (NMR)-based biomarker chips (Gaster et al, 2009) and micro-NMR devices (Haun et al, 2011) for the analysis of soluble biomarkers and biomarkers on whole cells, respectively. Because these devices, in contrast to currently used detection methods, are based on magnetic labeling and readout, they can analyze multiple biomarkers simultaneously without the need to purify samples before analysis.…”

Bioorthogonal chemistries for nanomaterial conjugation and targeting

“…Haun et al first used the TCO/Tz bioorthogonal coupling technique to amplify MNP binding to extracellular and intracellular cancer biomarkers for profiling tumor cells using a novel miniaturized diagnostic magnetic resonance device ( μ NMR) [47] . Tumor samples tested were obtained by fine-needle aspiration (FNA) biopsy from patients with suspected intra-abdominal tumors.…”