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Cell-surface sensors for real-time probing of cellular environments

Nature Nanotechnology volume 6, pages 524–531 (2011)Cite this article

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Abstract

The ability to explore cell signalling and cell-to-cell communication is essential for understanding cell biology and developing effective therapeutics. However, it is not yet possible to monitor the interaction of cells with their environments in real time. Here, we show that a fluorescent sensor attached to a cell membrane can detect signalling molecules in the cellular environment. The sensor is an aptamer (a short length of single-stranded DNA) that binds to platelet-derived growth factor (PDGF) and contains a pair of fluorescent dyes. When bound to PDGF, the aptamer changes conformation and the dyes come closer to each other, producing a signal. The sensor, which is covalently attached to the membranes of mesenchymal stem cells, can quantitatively detect with high spatial and temporal resolution PDGF that is added in cell culture medium or secreted by neighbouring cells. The engineered stem cells retain their ability to find their way to the bone marrow and can be monitored in vivo at the single-cell level using intravital microscopy.

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Figure 1: Mechanism of PDGF aptamer sensors in solution.
Figure 2: Anchoring the engineered aptamer sensor to the cell surface.
Figure 3: Aptamer sensor functions on the cell surface.
Figure 4: Spatial-temporal imaging of a single MSC functionalized with the quench sensor demonstrates that PDGF sensing correlates with data generated from a computational model.
Figure 5: Real-time sensing of PDGF secretion from neighbouring MDA-MB-231 cells by sensor-engineered MSCs.
Figure 6: Bone marrow homing and transmigration of aptamer-labelled MSCs.

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Acknowledgements

The authors thank Lei Xu and Dannie Wang for the preparation of PDGF-producing MDA-MB-231 cells. This work was supported by the National Institutes of Health (NIH; grants nos. HL097172, HL095722 and DE019191 to J.M.K., grants nos. HL082792 and NS059348 to J.L., and grant no. NIAID 5RC1AI086152 to J.C.L.), by the Charles A. Dana Foundation (J.C.L.) and by the American Heart Association (grant no. 0970178N to J.M.K.). W.Z. is supported by an International Human Frontier Science Program Organization postdoctoral fellowship. Y.J.Y. holds a National Science Foundation Graduate Fellowship.

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Authors and Affiliations

  1. Center for Regenerative Therapeutics & Department of Medicine, Brigham & Women's Hospital, 65 Landsdowne Street, Cambridge, 02139, Massachusetts, USA

    Weian Zhao, Sebastian Schafer, Joseph A. Phillips, Weisuong Teo, Ilia A. Droujinine, Cheryl H. Cui, Debanjan Sarkar & Jeffrey M. Karp

  2. Harvard Medical School, 65 Landsdowne Street, Cambridge, 02139, Massachusetts, USA

    Weian Zhao, Sebastian Schafer, Alicia L. Carlson, Joseph A. Phillips, Weisuong Teo, Ilia A. Droujinine, Cheryl H. Cui, Charles P. Lin, Debanjan Sarkar & Jeffrey M. Karp

  3. Harvard Stem Cell Institute, 65 Landsdowne Street, Cambridge, 02139, Massachusetts, USA

    Weian Zhao, Sebastian Schafer, Joseph A. Phillips, Weisuong Teo, Ilia A. Droujinine, Cheryl H. Cui, Charles P. Lin, Debanjan Sarkar & Jeffrey M. Karp

  4. Harvard-MIT Division of Health Science and Technology, 65 Landsdowne Street, Cambridge, 02139, Massachusetts, USA

    Weian Zhao, Sebastian Schafer, Joseph A. Phillips, Weisuong Teo, Ilia A. Droujinine, Cheryl H. Cui, Debanjan Sarkar & Jeffrey M. Karp

  5. Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, 02139, Massachusetts, USA

    Jonghoon Choi, Yvonne J. Yamanaka & J. Christopher Love

  6. Department of Biological Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, 02139, Massachusetts, USA

    Yvonne J. Yamanaka

  7. Department of Medicine, Brigham and Women's Hospital/Harvard Medical School, 65 Landsdowne Street, Cambridge, 02139, Massachusetts, USA

    Maria L. Lombardi & Jan Lammerding

  8. Department of Mechanical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, 02139, Massachusetts, USA

    Suman Bose & Rohit Karnik

  9. Wellman Center for Photomedicine and Center for Systems Biology, Massachusetts General Hospital/Harvard Medical School, 40 Blossom Street, Boston, 02114, Massachusetts, USA

    Alicia L. Carlson & Charles P. Lin

  10. Department of Radiation Oncology, Edwin L. Steele Laboratory, Massachusetts General Hospital/Harvard Medical School, 55 Fruit Street, Boston, 02114, Massachusetts, USA

    Rakesh K. Jain

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  1. Weian Zhao
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Contributions

W.Z. and J.M.K. are responsible for study concept and design. W.Z., J.M.K., J.L., J.C.L., C.P.L., D.S. and R.K. prepared the manuscript. W.Z., S.S., J.C., Y.J.Y., M.L.L., S.B., A.L.C., J.A.P, W.T., I.A.D. and C.C. carried out experiments and performed data analysis. R.K.J. provided genetically engineered cells.

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Correspondence to Jeffrey M. Karp.

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The authors declare no competing financial interests.

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Zhao, W., Schafer, S., Choi, J. et al. Cell-surface sensors for real-time probing of cellular environments. Nature Nanotech 6, 524–531 (2011). https://doi.org/10.1038/nnano.2011.101

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