title: Selective tracking of lysosomal Cu2+ ions using simultaneous target- and location-activated fluorescent nanoprobes.
abstract: Levels of lysosomal copper are tightly regulated in the human body. However, few methods for monitoring dynamic changes in copper pools are available, thus limiting the ability to diagnostically assess the influence of copper accumulation on health status. We herein report the development of a dual target and location-activated rhodamine-spiropyran probe, termed Rhod-SP, activated by the presence of lysosomal Cu(2+). Rhod-SP contains a proton recognition unit of spiropyran, which provides molecular switching capability, and a latent rhodamine fluorophore for signal transduction. Upon activation by lysosomal acidic pH, Rhod-SP binds with Cu(2+) by spiropyran-based proton activation, promoting, in turn, rhodamine ring opening, which shows a "switched on" fluorescence signal. However, to protect Rhod-SP from degradation and interference by the physiological environment, it is engineered on mesoporous silica nanoparticles (MSNs), and the surface of Rhod-SP@MSNs is further anchored with β-cyclodextrin (β-CD) to enhance the solubility and bioavailability of Rhod-SP@MSN-CD. Next, to enhance cell specificity, a guiding unit of c(RGDyK) peptide conjugated adamantane (Ad-RGD) as prototypical system, is incorporated on the surface of Rhod-SP@MSN-CD to target integrin αvβ3 and αvβ5 overexpressed on cancer cells. Fluorescence imaging showed that both Rhod-SP@MSN-CD and Rhod-SP@MSN-CD-RGD were suitable for visualizing exogenous and endogenous Cu(2+) in lysosomes of living cells. This strategy addresses some common challenges of chemical probes in biosensing, such as spatial resolution in cell imaging, the solubility and stability in biological system, and the interference from intracellular species. The newly designed nanoprobe, which allows one to track, on a location-specific basis, and visualize lysosomal Cu(2+), offers a potentially rich opportunity to examine copper physiology in both healthy and diseased states.
authors.affiliation: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Collaborative Innovation Center for Chemistry and Molecular Medicine, Hunan University , Changsha, 410082, China.
authors.forename: Yinhui
authors.initials: Y
authors.lastname: Li
authors.forename: Yirong
authors.initials: Y
authors.lastname: Zhao
authors.forename: Winghong
authors.initials: W
authors.lastname: Chan
authors.forename: Yijun
authors.initials: Y
authors.lastname: Wang
authors.forename: Qihua
authors.initials: Q
authors.lastname: You
authors.forename: Changhui
authors.initials: C
authors.lastname: Liu
authors.forename: Jing
authors.initials: J
authors.lastname: Zheng
authors.forename: Jishan
authors.initials: J
authors.lastname: Li
authors.forename: Sheng
authors.initials: S
authors.lastname: Yang
authors.forename: Ronghua
authors.initials: R
authors.lastname: Yang
mesh_terms: D001578:Benzopyrans / Q000737:chemistry; D058948:Cell Tracking / Q000379:methods*; D003300:Copper / Q000032:analysis*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007211:Indoles / Q000737:chemistry; D008247:Lysosomes / Q000737:chemistry / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D009574:Nitro Compounds / Q000737:chemistry; D012235:Rhodamines / Q000737:chemistry; D012822:Silicon Dioxide / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
chemical_list: D001578:Benzopyrans; D005456:Fluorescent Dyes; D007211:Indoles; D009574:Nitro Compounds; D012235:Rhodamines; C088184:spiropyran; D012822:Silicon Dioxide; D003300:Copper