Conformationally Induced Off–On Cell Membrane Chemosensor Targeting Receptor Protein-Tyrosine Kinases for in Vivo and in Vitro Fluorescence Imaging of Cancers

Abstract

Molecules capable of monitoring receptor protein-tyrosine kinase expression could potentially serve as useful tools for cancer diagnosis due to the overexpression of tyrosine kinases during tumor growth and metastasis. In this work, a conformationally induced "off-on" tyrosine kinase cell membrane fluorescent sensor (SP1) was designed and evaluated for the detection and imaging of receptor protein-tyrosine kinases in vivo and in vitro. SP1 consists of sunitinib and pyrene linked via hexamethylenediamine and displays quenched fluorescence as a dimer. The fluorescence of SP1 is restored in the presence of receptor protein-tyrosine kinases upon strong interaction with SP1 at the target terminal. The unique signal response mechanism enables SP1 use for fluorescence microscopy imaging of receptor protein-tyrosine kinases in the cell membranes of living cells, allowing for the rapid differentiation of cancer cells from normal cells. SP1 can be used to visualize the chick embryo chorioallantoic membrane and mouse model tumors, suggesting its possible application for early cancer diagnosis.

Dissection of Protein Kinase Pathways in Live Cells Using Photoluminescent Probes: Surveillance or Interrogation?

“…This enrichment can be distinguished from the background either after washout of the probe, or by virtue of change of photoluminescence properties of the probe associated with its binding to the target PK. Importantly, in recent years, use of Class 3A and Class 3B probes for theranostics and fluorescence-guided surgeries has been extensively explored [82,83,162,163].…”

“…Different approaches have been used to overcome this issue, including application of cell membrane-penetrating peptides [171] and derivatization of probes with functional groups that are cleaved by intracellular enzymes upon delivery of the probe into live cells (so-called prodrug strategy [172]). Probes utilizing smaller hydrophobic sensing fragments and non-polar fluorophores can cross cell plasma membrane on their own [83,84,163,173], although there is a risk that the probe is recruited into the hydrophobic layer instead of reaching target PK in the cytoplasm. In certain cases, assembly of probe in cellulo has been reported, where the probe was first sent into cells, followed by application of fluorophore containing selective reactive moiety [174,175].…”

“…However, fixation is associated with disruption of membrane structures, which can give rise to significant artifacts in imaging: the localization of the probe before and after fixation may differ drastically. Therefore, importance of fluorogenic dyes and probes that gain photoluminescence upon binding to PK has been recognized in recent years [72,84,163,[182][183][184].…”

Rutin-modified silver nanoparticles as a chromogenic probe for the selective detection of Fe3+in aqueous medium

“…9,10 In the last decade, nanoscaled materials have been reported as selective probes for metal ions, including Fe 3+ . [11][12][13][14][15][16][17][18][19][20][21][22][23][24] Silver nanoparticles (AgNPs) are of particular interest because of the affordable price of starting materials, ease of controlling size and morphology, possibility to functionalize their surface with organic molecules, and optical properties that enable detection of a variety of analytes via simple UV-vis spectroscopy and digital image analysis. Furthermore, applications of AgNPs are also biotechnologically relevant due to the possibility of green synthetic protocols, including the use of plant extracts, 25 natural sources, 26 glycerol, 27 among others.…”

Optical Imaging of the Nanoscale Structure and Dynamics of Biological Membranes

“…This bipolar structure is specifically localized in the plasma membrane.Initially it has a negligible fluorescence, but upon binding with nitric oxide its fluorescence is enhanced. In another study, a conformationally induced off-on probe was developed by linking hexamethylenediamine with sunitinib and pyrene to target and image the tyrosine kinase receptor protein in cell membranes 134. The TLSHalo probe was developed to image the potassium ion transportation in cell membranes 135.…”