Abstract
This study describes the development of targeted quantum dots (T-QDs) as biomarkers for the labeling of glial progenitor cells (GPCs) that over express platelet derived growth factor (PDGF) and its receptor PDGFR (GPCPDGF). PDGFR plays a critical role in glioma development and growth, and is also known to affect multiple biological processes such as cell migration and embryonic development. T-QDs were developed using streptavidin-conjugated quantum dots (S-QDs) with biotinylated antibodies and utilized to label the intracellular and extracellular domains of live, cultured GPCPDGF cells via lipofection with cationic liposomes. Confocal studies illustrate successful intracellular and extracellular targeted labeling within live cells that does not appear to impact upstream PDGFR dynamics during real-time signaling events. Further, T-QDs were nontoxic to GPCPDGF cells, and did not alter cell viability or proliferation over the course of 6 days. These results raise new applications for T-QDs as ultra sensitive agents for imaging and tracking of protein populations within live cells, which that will enable future mechanistic study of oncogenic signaling events in real-time.
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References
Ballou B., C.B. Lagerholm, L.A. Ernst, M.P. Bruchez and A.S. Waggoner. Non invasive imaging of quantum dots in mice. Bioconjugate Chem. 15: 79-86, 2004.
Barber K., R.R. Mala, M.P. Lambert, R. Qiu, R.C. MacDonald and W.L. Klein. Delivery of membrane-impermeant fluorescent probes into living neural cell populations by lipotransfer. Neurosci. Lett. 207:17–20, 1996.
Chalmers, N. I., R. J. Palmer Jr., L. Du-Thumm, R. Sullivan, W. Shi, and P. E. Kolenbrander. Use of quantum dot luminescent probes to achieve single cell resolution of human oral bacteria in biofilms. Appl. Environ. Microbiol. 73:630–636, 2007.
Dai C., J. Celestino, Y. Okada, D.N. Louis, G.N. Fuller and E.C. Holland. PDGF autocrine stimulation dedifferentiates cultured astrocytes and induces oligodendrogliomas and oligoastrocytomas from neural progenitors and astrocytes in vivo. Genes Dev. 15:1913-1925, 2001.
Dai C. and E.C. Holland. Astrocyte differentiation states and glioma formation. Cancer J. 9: 72-81, 2003.
Dai C., Y. Lyustikman, A. Shih, X. Hu, G.N. Fuller, M. Rosenbulm and E.C. Holland. The characterstic of astrocytomas and oligodendrogliomas are caused by two distinct and interchangable signaling formats. Neoplasia. 7: 397-406, 2005.
Derfus A.M., W.C.W.Chan and S.N.Bhatia. Intracellular delivery of quantum dots for live cell labeling and organelle tracking. Adv. Mater. 16: 961-966, 2004.
Derfus A.M., W.C.W. Chan and S.N. Bhatia. Probing the cytotoxicity of semiconductor quantum dots. Nano Lett. 4: 11-18, 2004.
Dudu V., M. Ramcharan, L. Gilchrist, E.C. Holland and M.Vazquez. Liposome delivery of quantum dot to the cytosol of live cell. J Nanosci. Nanotechnol. 8: 2293-2300, 2008.
Gao X, Y. Cui, R.M.Leveson, L.W.K. Chung and S. Nie. In vivo cancer targeting and imaging with semiconductor quantum dots. Nat. Biotechnol. 22: 969-976, 2004.
Heldin C.H. and B. Westermark. Mechanism of action and in vivo role of platelet derived growth factor. Physiol. Rev. 79:1283-1316, (1999).
Holland E.C., Y. Li, J. Celestino, C. Dai, L. Schaefer, R. Sawaya and G.N. Fuller. Astrocytes give rise to oligodendrogliomas and astrocytomas after gene transfer of polyoma virus middle T antigen in vivo. Am. J. Pathol. 157:1031-1037, 2000.
Ipe I.B., A.Shukla, H.Lu, B.Zou, H. Rehage and C.M. Niemeyer. Dynamic light-scattering analysis of the electrostatic interaction of hexahistidine-tagged cytochrome P450 enzyme with semiconductor quantum dots. Chem. Phys. Chem. 7: 1112-1118, 2006.
Jaiswal J.K., H. Mattoussi, J.M. Mauro and S.M. Simon. Long term multiple color imaging of live cells using quantum dot bioconjugates. Nat. Biotechnol. 21: 47-51, 2003.
Jaiswal J.K. and S.M Simon. Potentials and pitfalls of fluorescent quantum dots for biological imaging. Trends Cell Biol. 14: 497-504, 2004.
Jones A.V. and N.C.P. Cross. Oncogenic derivatives of platelet-derived growth factor receptors. Cell. Mol. Life Sci., 61:2912-2923, 2004.
Jovin T.M. Quantum dots finally come of age. Nat. Biotechnol. 21:32-33, 2003.
Lasic D.D. Liposomes: From Physics to Applications. Elsevier Science Publishers BV, the Netherlands, 1993.
Liu H.Y. and T.Q. Vu. Identification of quantum dot bioconjugates and cellular protein co-localization by hybrid gel blotting. Nano Lett. 7: 1044-1049, 2007.
Lokker N.A., C.M. Sullivan, S.J. Hollenbach, M.A Israel and N.A. Giese. Platelet-derived growth factor (PDGF) autocrine signaling regulates survival and mitogenic pathways in glioblastoma cells: evidence that the novel PDGF-C and PDGF-D ligands play a role in development of brain tumors. Cancer Res. 62: 3729-3735, 2002.
Rajan S.S. and T.Q. Vu. Quantum Dots Monitor TrkA Receptor Dynamics in the Interior of Neural PC12 Cells. Nano Lett. 6:2049-2059, 2006.
Shih, A.H. and E.C. Holland. Platelet derived growth factor (PDGF) and glial tumorigenesis. Can. Lett. 232: 139-147, 2006.
Vu, T. Q., S. S. Rajan, and H. Y. Liu. Ligand bound quantum dots for intracellular imaging of neural receptors. In: Proc. SPIE, Colloidal Quantum Dots for Biomedical Applications, 2007, p. 6448.
Yu J., C. Ustach and H.R.C. Kim. Platelet derived growth factor signaling and human cancer. J. Biochem. Mol. Biol. 36: 49-59, 2003.
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This study was supported by grants from NSF CBET 0428573, NIH R21 CA118255, and PSC-CUNY.
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Sabharwal, N., Holland, E.C. & Vazquez, M. Live Cell Labeling of Glial Progenitor Cells Using Targeted Quantum Dots. Ann Biomed Eng 37, 1967–1973 (2009). https://doi.org/10.1007/s10439-009-9703-4
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DOI: https://doi.org/10.1007/s10439-009-9703-4