A fluorogenic probe for predicting treatment response in non-small cell lung cancer with EGFR-activating mutations

Abstract

Therapeutic responses of non-small cell lung cancer (NSCLC) to epidermal growth factor receptor (EGFR) - tyrosine kinase inhibitors (TKIs) are known to be associated with EGFR mutations. However, a proportion of NSCLCs carrying EGFR mutations still progress on EGFR-TKI underlining the imperfect correlation. Structure-function-based approaches have recently been reported to perform better in retrospectively predicting patient outcomes following EGFR-TKI treatment than exon-based method. Here, we develop a multicolor fluorescence-activated cell sorting (FACS) with an EGFR-TKI-based fluorogenic probe (HX103) to profile active-EGFR in tumors. HX103-based FACS shows an overall agreement with gene mutations of 82.6%, sensitivity of 81.8% and specificity of 83.3% for discriminating EGFR-activating mutations from wild-type in surgical specimens from NSCLC patients. We then translate HX103 to the clinical studies for prediction of EGFR-TKI sensitivity. When integrating computed tomography imaging with HX103-based FACS, we find a high correlation between EGFR-TKI therapy response and probe labeling. These studies demonstrate HX103-based FACS provides a high predictive performance for response to EGFR-TKI, suggesting the potential utility of an EGFR-TKI-based probe in precision medicine trials to stratify NSCLC patients for EGFR-TKI treatment.

Small-molecule probes from bench to bedside: advancing molecular analysis of drug–target interactions toward precision medicine

“…60 Moreover, through structural modification, these probes can be fine-tuned to achieve various electronic properties, resulting in improved brightness, lifetime, photo-stability, and excitation/emission wavelengths, as well as environmental sensitivity and activation mechanisms. 61–64…”

“…60 Moreover, through structural modification, these probes can be finetuned to achieve various electronic properties, resulting in improved brightness, lifetime, photo-stability, and excitation/ emission wavelengths, as well as environmental sensitivity and activation mechanisms. [61][62][63][64] For biomedical applications, organic fluorophores such as fluorescein (12), rhodamine (13), cyanine (14) and boron dipyrromethene (BODIPY, 15) are widely used due to their good photostability, high quantum yield and excellent hydrophilicity. 65 Xanthene dyes like fluorescein and rhodamine derivatives exhibit absorption and emission in the range of 500-600 nm, and exist in a dynamic equilibrium between a cell-permeable, non-fluorescent spirolactone and a highly soluble, fluorescent zwitterion.…”

Environment-Sensitive Fluorescent Probe Enables Assessment of Anaplastic Lymphoma Kinase Activity in Nonsmall Cell Lung Cancer

“…29,30 For example, environment-sensitive fluorophores [e.g., 4-sulfonamidebenzoxadiazole (SBD) 31 have unique emission properties that are highly sensitive to the immediate environment, thereby showing "turn-on" properties. 32 Specifically, the environmentsensitive probes usually show weak fluorescence in the polar and protic environment, while they can be activated when in hydrophobic surroundings. Since protein-binding sites are often very hydrophobic, the environment-sensitive probes with fluorescence "turn-on" properties thus can be achieved by integrating an environment-sensitive fluorophore and a protein-targeted ligand (e.g., ALK-TKI).…”

N-Functionalized fluorophores: detecting urinary albumin and imaging lipid droplets

“…In addition to its applications in medicinal chemistry, the sulfonyl group also plays a critical role in creating various luminescent platforms, such as Texas Red-X and Rhodamine Red-X. 7 A few other examples include squaraine-sulfonamide derivatives for NIR imaging, 8 the 1,8-naphthalimide-4-sulfonamide-linked kinase inhibitor crizotinib for the interaction with GSH, 9 tracking glutathione levels within lysosomes, 10 hypoxia imaging by covalently linking fluorophores with carbonic anhydrase inhibitors, 11 sulfonamidyl luciferins, 12 multicolor fluorescent ESIPT fluorophores, 13 rhodamine derivatives for super-resolution imaging, 14 Golgi-targeting, 15,16 profiling active-EGFR in tumors, 16 locating GPR120 receptors, 17 endoplasmic reticulum (ER)-targeting pendants [18][19][20][21][22] and imaging the lipid droplets using mono dansylpentane (MDH). 23 In all these cases, the sulfonyl group exists as sulphonamide (R-S(vO) 2 -NH 2 ), where the NH hydrogen links functional elements of interest.…”