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Controlled drug release by a nanorobot
Jinglin Fu & Hao Yan
A tiny, locked box made of DNA opens up to release drug molecules in the presence of target cells.
In the 1966 film Fantastic Voyage, a shrunken submarine is injected into a man’s circulatory system to find and destroy a lifethreatening blood clot in his brain. Scientists have embraced this science-fictional concept in the form of research on targeted or ‘smart’ drug delivery: nano- or microscale systems that can discriminate between healthy and diseased cells and selectively deliver medicinal payloads. In a recent paper in Science, Douglas et al.1 describe a nanoscale DNA cage that releases Fab antibody fragments in the pre sence of target cells in vitro. Although the utility of this approach for targeted drug delivery in vivo remains unclear, the study shows how DNA nanostructures can be combined with rudimentary robotic functions to induce cell signaling pathways. The work of Douglas et al.1 builds on advances in DNA nanotechnology that allowed the construction of sophisticated multidimensional structures2 and of devices capable of robot-like functions such as molecular sensing3, logical computation4 and activation5. The authors produced their nanostructures using ‘DNA origami’, an approach for creating two- or three-dimensional nanoscale shapes by folding a long singlestranded DNA molecule along a predetermined path using oligonucleotide ‘staples’6. Nanostructures made by DNA origami can be designed to present addressable surface features at which other particles or molecules can be precisely positioned2. This capability has been used to generate simple twodimensional structures that direct the motion of robots constructed from DNAzymes 7. Several years ago, a report describing a three-dimensional DNA box with a lid that can be opened by a DNA ‘key’ showed that DNA-origami structures are capable of acting as dynamic containers8. Douglas et al.1 extended this line of research by designing a box that releases its cargo in the presence of a specific configuration of target molecules. The barrel-shaped device, which the authors call a “nanorobot” or “nanobot”,
Jinglin Fu and Hao Yan are in the Department of Chemistry and Biochemistry and the Biodesign Institute at Arizona State University, Tempe, Arizona, USA. e-mail: firstname.lastname@example.org
consists of two halves connected by a switchable hinge (Fig. 1). Two distinct DNA aptamers are used to close and lock the DNA barrel. Because each of the aptamers specifically binds different protein antigens, they form an AND logic gate that requires the presence of two protein targets to be activated. When both aptamers bind their targets, a conformational change in the barrel releases the cargo. Thus, the aptamer-encoded lock functions as a sense-compute-actuate mechanism that could in principle be deployed to trigger a specific therapeutic response. In a first set of experiments, Douglas et al.1 used their aptamer-gated DNA nanobots to detect selected biomarkers (such as plateletderived growth factor or protein tyrosine kinase 7) expressed on the surface of leukemia cells. Several combinations of three aptamers that recognize different surface antigens were incorporated into the nanobot. To characterize the system, the authors used fluorescently labeled Fab antibody fragments as the payload; the nanobots showed highly specific binding to the cells that displayed the correct combination of surface antigens, even in mixed populations of whole-blood leukocytes. Next, the nanobots were used to activate signaling pathways in target cells. The authors loaded the nanobots with Fab antibody fragments known to bind human CD33 and human CDw328 and induce growth arrest in leukemic cells. Upon recognition of the surface antigen PDGF on cells from a patient with aggressive lymphocytic NK-type leukemia, the barrel structure opened, allowing the antibody payload
to bind to cell-surface receptors and inhibit the growth of the target cells. Similarly, an increase in T cell activation was induced by a nanobot loaded with Fab fragments specific for human CD3ε and flagellin. Additional measurements that quantify the AND gates’ sensitivity to the inputs, the rate of erroneous nanobot activation and the correlation between the number of activated nanobots with successfully modified cell signaling pathways would be required to fully evaluate nanobot performance. The aim of smart drug-delivery systems is to administer smaller drug doses to patients while offering improved therapeutic efficiency and fewer side effects compared with conventional drug delivery methods. Notable recent developments have included systems based on liposomes, polymersomes, micelles, nanoparticles and antibodies9. Yet these approaches do not provide the wide range of design modularity and structural programmability that DNA nanotechnology affords. Much work remains before the approach of Douglas et al.1 could be used for drug delivery in vivo. For example, nanobots might not be stable in the presence of nucleases and other enzymes in the blood. Increased resistance to degradation may be achievable by methods such as chemical cross-linking of selected DNA strands or the use of peptide nucleic acids or locked nucleic acids. Similarly, aptamerencoded locks may lose specificity and efficiency in protein-rich serum. In addition, delivery into cells is required for broad application of a drug-delivery system
© 2012 Nature America, Inc. All rights reserved.
Short staple strands
Antigens unlock aptamers
Exposed payload triggers signaling responses in cells
Figure 1 DNA nanobot for targeted drug delivery. To fabricate the nanobot by DNA origami, a long single-stranded DNA scaffold is folded into the shape of a hexagonal barrel by short oligonucleotide ‘staple’ strands. The assembled nanobot consists of two aptamer-encoded locks, a barrel-shaped body and a bound molecular cargo. The use of two different aptamers that ‘unlock’ when exposed to two antigens results in an AND logic gate, meaning that the nanobot opens in the presence of the correct combination of antigen keys. The molecular payload is then released to bind to target cells and activate signaling pathways.
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Single-stranded bacteriophage DNA
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COMPETING FINANCIAL INTERESTS The authors declare no competing financial interests.. Hou et al. 8. et al. UK.ne w s and v i e w s
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Carlos Caldas is in the Department of Oncology at the University of Cambridge and at Cancer Research UK.3 and in the New England Journal of Medicine4.. W. S.. Delebecque.M.3 carried out exome sequencing on single cells from an essential thrombocythemia tumor and a kidney tumor. P. 1196–1201 (2011). In both cases. J. Hou et al. WGA DNA was then analyzed by massively parallel sequencing with Illumina instruments. Pinheiro.
Cancer sequencing unravels clonal evolution
Characterization of tumor heterogeneity at the sequence level presents new challenges and opportunities for targeted therapies.J. W..
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. of which 171 coding variants were further assessed. 3. to a mean depth of 30×.E. metastasis and resistance to therapy1 (Fig.S.uk
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