Kinetic Cellular Phenotypic Profiling: Prediction, Identification, and Analysis of Bioactive Natural Products

Abstract

Natural products have always been a major source of therapeutic agents; however, the development of traditional herbal products has been currently hampered by the lack of analytic methods suitable for both high-throughput screening and evaluating the mechanism of action. Cellular processes such as proliferation, apoptosis, and toxicity are well-orchestrated in real time. Monitoring these events and their perturbation by natural products can provide high-rich information about cell physiological relevancies being involved. Here, we report a novel cell-based phenotypic profiling strategy that uses electronic impedance readouts for real-time monitoring of cellular responses to traditional Chinese medicines (TCMs). The utility of this approach was used to screen natural herbs that have been historically documented to cure human diseases and that have been classified into seven clusters based on their mechanisms of action. The results suggest that herbal medicines with similar cellular mechanisms produce similar time/dose-dependent cell response profiles (TCRPs). By comparing the TCRPs produced by the Chinese medicinal Cordyceps sinensis with similar TCRPs of chemical compounds, we explored the potential use of herbal TCRPs for predicting cellular mechanisms of action, herbal authentications, and bioactive identification. Additionally, we further compared this novel TCRP technology with high-performance liquid chromatography (HPLC)-based methods for herbal origin-tracing authentication and identification of bioactive ingredients. Together, our findings suggest that using TCRP as an alternative to existing spectroscopic techniques can allow us to analyze natural products in a more convenient and physiologically relevant manner.

Phenotypic Pattern-Based Assay for Dynamically Monitoring Host Cellular Responses to Salmonella Infections

“…Cell-based TCRP technology that evaluates multiple physiological changes would provide richer information about interactions between bacteria and hosts than endpoint assays [14], [41], [42]. However, the inherent variability and complexity of cells should be taken into full consideration in experimental design, implementation, and analysis.…”

“…However, the inherent variability and complexity of cells should be taken into full consideration in experimental design, implementation, and analysis. Introduction of statistical model of TCRPs will address this problem [41]. In addition, because of the ability to measure over time and dosage, TCRP offers the possibility of using multiple variables in mathematic simulations for kinetic measurement of cell phenotypic changes.…”

Label-free drug discovery

“…The most popular is to profile compound-induced cell signaling in confluent cells, given that the cells once reach confluency start to enter a new growth cycle or a quiescent state and the compound-induced response is almost exclusively due to cell signaling (Fang, 2010, 2011a). Alternatively, the long-term impacts of compounds on cell growth can also be used to screen compound library (Abassi et al, 2009; Fu et al, 2011). Of note, this approach may be able to identify ligands for other classes of targets such as nuclear receptors whose activation by themselves may not result in rapid signaling-related biosensor responses.…”

Review on Pharmacologically Active Metabolites from Yarsagumba (Ophiocordyceps sinensis), an Epitome of Himalayan Elixir

“…The uses of multiple chemical markers such as nucleosides, ergosterol, mannitol and polysaccharides used for quality control also have some drawbacks (Li et al 2006a). Fu et al (2011) reported process for more precise authentication of sample continuous monitoring of cellular impedance in real time, which produces specific time/dose dependent cell response profiles (TCRPs) in addition to spectroscopic fingerprinting of active compounds by HPLC using adenosine (or 3-deoxyadenosine) as a standard, in accordance with Pharmacopoeia of the People's Republic of China (2005 version). The use of bar-coding and fingerprinting to some extent are in practice now-a-days with accession of ITS and 18s ribosomal RNA gene (MMDBD 2012).…”