Light-up bioprobe with aggregation-induced emission characteristics for real-time apoptosis imaging in target cancer cells.

Specific bioprobes that are capable of real-time and targeted monitoring and imaging of cancer cell apoptosis are highly desirable for cancer diagnosis and the evaluation of cancer therapy efficacy. In this work, an asymmetric fluorescent light-up bioprobe with aggregation-induced emission (AIE) characteristics was designed and synthesized by the conjugation of two different hydrophilic peptides, caspase-specific Asp-Glu-Val-Asp (DEVD) and cyclic Arg-Gly-Asp (cRGD), onto a typical AIE luminogen of a tetraphenylsilole (TPS) unit. The asymmetric probe is almost non-emissive in aqueous solution and its fluorescence is significantly switched on in the presence of caspase-3. The fluorescence turn-on is due to the cleavage of the DEVD moiety by caspase-3, and the aggregation of released TPS-cRGD residues, which restricts the intramolecular rotations of TPS phenyl rings and populates the radiative decay channels. Application of the asymmetric light-up probe for real-time targeted imaging of cancer cell apoptosis is successfully demonstrated using integrin αvβ3 receptor overexpressing U87MG human glioblastoma cells as an example. The probe shows specific targeting capability to U87MG cancer cells by virtue of the efficient binding between cRGD and integrin αvβ3 receptors and is able to real-time monitor and image cancer cell apoptosis in a specific and sensitive manner.

[1]  Akira Kanno,et al.  Cyclic luciferase for real-time sensing of caspase-3 activities in living mammals. , 2007, Angewandte Chemie.

[2]  S. Fukushima,et al.  Cyclic RGD Peptide-Conjugated Polyplex Micelles as a Targetable Gene Delivery System Directed to Cells Possessing αvβ3 and αvβ5 Integrins , 2007 .

[3]  C. Roy,et al.  Inhibition of pathogen-induced apoptosis by a Coxiella burnetii type IV effector protein , 2010, Proceedings of the National Academy of Sciences.

[4]  Yigong Shi,et al.  Molecular mechanisms of caspase regulation during apoptosis , 2004, Nature Reviews Molecular Cell Biology.

[5]  D. Hanahan,et al.  The Hallmarks of Cancer , 2000, Cell.

[6]  Ben Zhong Tang,et al.  Fluorescent bio/chemosensors based on silole and tetraphenylethene luminogens with aggregation-induced emission feature , 2010 .

[7]  Ralph Weissleder,et al.  A fluorescent nanosensor for apoptotic cells. , 2006, Nano letters.

[8]  Kristin L. Hazelwood,et al.  A bright and photostable photoconvertible fluorescent protein for fusion tags , 2009, Nature Methods.

[9]  R. Hebbel,et al.  Internucleosomal cleavage of DNA as the sole criterion for apoptosis may be artifactual. , 1994, The Journal of laboratory and clinical medicine.

[10]  B. Tang,et al.  Bifunctional combined Au-Fe(2)O(3) nanoparticles for induction of cancer cell-specific apoptosis and real-time imaging. , 2012, Biomaterials.

[11]  Ben Zhong Tang,et al.  Real-time monitoring of cell apoptosis and drug screening using fluorescent light-up probe with aggregation-induced emission characteristics. , 2012, Journal of the American Chemical Society.

[12]  A. Porter,et al.  Emerging roles of caspase-3 in apoptosis , 1999, Cell Death and Differentiation.

[13]  R. Davis,et al.  In vivo detection and imaging of phosphatidylserine expression during programmed cell death. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[14]  H S Kwok,et al.  Aggregation-induced emission of 1-methyl-1,2,3,4,5-pentaphenylsilole. , 2001, Chemical communications.

[15]  Ben Zhong Tang,et al.  Aggregation-induced emission. , 2011, Chemical Society reviews.

[16]  Igor L. Medintz,et al.  Sensing caspase 3 activity with quantum dot-fluorescent protein assemblies. , 2009, Journal of the American Chemical Society.

[17]  Nai-Tzu Chen,et al.  The protease-mediated nucleus shuttles of subnanometer gold quantum dots for real-time monitoring of apoptotic cell death. , 2010, Journal of the American Chemical Society.

[18]  Ben Zhong Tang,et al.  Biocompatible Nanoparticles with Aggregation‐Induced Emission Characteristics as Far‐Red/Near‐Infrared Fluorescent Bioprobes for In Vitro and In Vivo Imaging Applications , 2012 .

[19]  S. Diamond.  Methods for mapping protease specificity. , 2007, Current opinion in chemical biology.

[20]  I. Kwon,et al.  Small heat shock protein as a multifunctional scaffold: integrated tumor targeting and caspase imaging within a single cage. , 2011, Biomacromolecules.

[21]  B. Tang,et al.  Fluorescent chemosensor for detection and quantitation of carbon dioxide gas. , 2010, Journal of the American Chemical Society.

[22]  Wenlin Huang,et al.  Simultaneous modulation of COX‐2, p300, Akt, and Apaf‐1 signaling by melatonin to inhibit proliferation and induce apoptosis in breast cancer cells , 2012, Journal of pineal research.

[23]  Kwangmeyung Kim,et al.  Multiplex imaging of an intracellular proteolytic cascade by using a broad-spectrum nanoquencher. , 2012, Angewandte Chemie.

[24]  S. Korsmeyer,et al.  Cell Death in Development , 1999, Cell.

[25]  Sanjiv S. Gambhir,et al.  Near-Infrared Fluorescent RGD Peptides for Optical Imaging of Integrin αvβ3 Expression in Living Mice , 2005 .

[26]  J. Lovell,et al.  Porphyrin FRET acceptors for apoptosis induction and monitoring. , 2011, Journal of the American Chemical Society.

[27]  B. Tang,et al.  Aggregation-Induced Emission of Tetraarylethene Luminogens , 2010 .

[28]  S. Nagata,et al.  Apoptosis by Death Factor , 1997, Cell.

[29]  A. Miyawaki,et al.  A high-throughput method for development of FRET-based indicators for proteolysis. , 2004, Biochemical and biophysical research communications.

[30]  Yaokai Gan,et al.  The use of autologous enriched bone marrow MSCs to enhance osteoporotic bone defect repair in long-term estrogen deficient goats. , 2012, Biomaterials.

[31]  Kwangmeyung Kim,et al.  Real time, high resolution video imaging of apoptosis in single cells with a polymeric nanoprobe. , 2011, Bioconjugate chemistry.

[32]  Ben Zhong Tang,et al.  Aggregation-induced emission: phenomenon, mechanism and applications. , 2009, Chemical communications.

[33]  M. Pooga,et al.  Cell-penetrating Peptides Split into Two Groups Based on Modulation of Intracellular Calcium Concentration* , 2012, The Journal of Biological Chemistry.

[34]  Hao Hong,et al.  Cancer-targeted optical imaging with fluorescent zinc oxide nanowires. , 2011, Nano letters.

[35]  Ick Chan Kwon,et al.  Cell-permeable and biocompatible polymeric nanoparticles for apoptosis imaging. , 2006, Journal of the American Chemical Society.

[36]  Igor Jurisica,et al.  Hypoxia predicts aggressive growth and spontaneous metastasis formation from orthotopically grown primary xenografts of human pancreatic cancer. , 2011, Cancer research.

[37]  B. Tang,et al.  Aggregation-Induced Emission and Biological Application of Tetraphenylethene Luminogens , 2011 .

[38]  Ben Zhong Tang,et al.  Specific detection of integrin αvβ3 by light-up bioprobe with aggregation-induced emission characteristics. , 2012, Journal of the American Chemical Society.

[39]  Kai Li,et al.  Photostable fluorescent organic dots with aggregation-induced emission (AIE dots) for noninvasive long-term cell tracing , 2013, Scientific Reports.

[40]  R. Moats,et al.  In vivo Near-Infrared Fluorescence Imaging of Integrin αvβ3 in Brain Tumor Xenografts , 2004, Cancer Research.

[41]  W. Melhuish,et al.  QUANTUM EFFICIENCIES OF FLUORESCENCE OF ORGANIC SUBSTANCES: EFFECT OF SOLVENT AND CONCENTRATION OF THE FLUORESCENT SOLUTE1 , 1961 .

[42]  Jong Seung Kim,et al.  Direct fluorescence monitoring of the delivery and cellular uptake of a cancer-targeted RGD peptide-appended naphthalimide theragnostic prodrug. , 2012, Journal of the American Chemical Society.

[43]  Zhuang Liu,et al.  Drug delivery with carbon nanotubes for in vivo cancer treatment. , 2008, Cancer research.