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Published in final edited form as: Adv Mater. 2015 Sep 29;27(44):7043–7050. doi: 10.1002/adma.201503323

Anticancer platelet-mimicking nanovehicles

Quanyin Hu 1,2, Wujin Sun 3,4, Chengen Qian 5,6, Chao Wang 7,8, Hunter N Bomba 9,10, Zhen Gu 11,12,13,*
PMCID: PMC4998740  NIHMSID: NIHMS735237  PMID: 26416431

Biomimetic drug delivery system offers new opportunities to mimic the biological particulates, including cells, vesicles and viruses for enhancing biocompatibility and promoting therapeutic efficacy.[1] As a simple and effective biomimetic approach, delivery vehicles coated with cell membranes are currently being intensely pursued to achieve a variety of merits, such as prolonging circulation time, alleviating immunogenicity and achieving active targeting ability. Typical examples include red blood cell (RBC) membrane-coated PLGA nanoparticles,[2] white blood cell (WBC) membrane-decorated silica particles[3] and cancer cell membrane-cloaked nanoparticles.[4] Given the complexity of biological entities with different sorts of membranes integrated with distinct bioactive components, versatile biomimetic drug delivery systems with high specificity are expected to develop. Platelet is an indispensable component of blood stream with the ability of targeting vascular injury sites to impede thrombogenesis and maintaining the integrity of blood circulation.[5] Recently, the recognition and interaction between platelets and circulating tumor cells in blood have aroused considerable attention because of its crucial contribution to tumor metastasis.[6] The aggregation of platelets surrounding circulating tumor cells (CTCs) helps CTCs survive in blood stream and spread to new tissues.[7] The mechanism underlying this specific aggregation includes biomolecular binding such as P-Selectin and CD44 receptors[8] and structure-based capture.[9]

Here we report the development of platelets membrane (PM)-coated core-shell nanovehicle (designated PM-NV) for targeting and sequential and site-specific delivery of extracelluarly active protein and intracellular functional small molecular drug. As displayed in Figure 1a, the PM-NV is composed of two components: 1) a nanogel based inner core part for loading small molecular drug; 2) the platelet membranes based outer shell for decoration of protein drug. The core NV is made by a single emulsion method and crosslinked by an acid-degradable crosslinker. The platelet membranes are derived from platelets and purified to coat the surface of NV. Equipped with large numbers of “self-recognized” proteins, PM-NV is expected to minimize the in vivo immunogenicity, prolong the circulation time.[10] More importantly, the overexpressed P-Selectin on the PM can specifically bind to CD44 receptors upregulated on the surface of cancer cells. Taken together, we hypothesized the PM-NV could actively target to tumor site and sequentially deliver anticancer therapeutics to their most active destinations. To demonstrate our hypothesis, we loaded two anticancer therapeutics—TRAIL and Dox into PM-NV (designated TRAIL-Dox-PM-NV). As one of the most important extracellular activators of apoptosis, TRAIL induces apoptosis of tumor cells by binding to the death receptors (DR4, DR5) on the cell surface;[11] while Dox can intercalate the nuclear DNA of cancer cells to trigger the intrinsic apoptosis signaling pathway.[12]

Figure 1.

Figure 1

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Schematic design of drug-loaded PM-NV for targeting and sequential drug delivery. a). The main components of TRAIL-Dox-PM-NV: TRAIL-conjugated platelets membrane derived from platelets; Dox-loaded nanovehicle (Dox-NV). I: centrifugation of whole blood; II: isolation of platelets; III: extraction of platelets membrane. b), In vivo elimination of circulating tumor cells (CTCs) and sequential delivery of TRAIL and Dox. TRAIL-Dox-PM-NV captured the CTCs via specific affinity of P-Selectin and overexpressed CD44 receptors and subsequently triggered TRAIL/Dox-induced apoptosis signaling pathways. I: the interaction of TRAIL and death receptors (DRs) to trigger the apoptosis signaling; II: the internalization of TRAIL-Dox-PM-NV; III: the dissociation of TRAIL-Dox-PM-NV mediated by the acidity of lyso-endosome; IV: release and accumulation of Dox in the nuclei; V: intrinsic apoptosis triggered by Dox.

After intravenous (i.v.) injection, the PM-NVs are expected to accumulate in the tumor site by combination of the passive enhanced permeability and retention (EPR) effect[13] and active targeting based on the affinity between PM and overexpressed CD44 receptors on the cancer cells (Figure 1b). Meanwhile, the aggregation of PM-NV on the surface of cancer cells enabled by capture ability of P-Selectin could facilitate the interaction between TRAIL and cell membranes and subsequently initiation of extrinsic apoptosis signaling. Additionally, the affinity of P-Selectin and CD44 are also expected to readily eliminate CTCs (Figure 1b), which plays a vital role in tumor metastasis.[14] After cellular internalization, the acidity in the endo-lysosome is expected to digest the acid-responsive modality in the PM-NV, accompanied by the release of encapsulated Dox, which will accumulate in the nuclei of cancer cells for synergistically inducing apoptosis.

In order to prepare TRAIL-Dox-PM-NV, the purified platelets were collected through the gradient centrifugation.[15] The obtained platelets were verified by fluorescence-activated cell sorting (FACS) study through labeling with antibodies against the platelet-specific marker, CD41,[16] and a marker for white blood cells (WBC), CD45.[17] Strong fluorescence signals associated with CD41 were observed and signals associated with CD45 were barely observed (Figure S1), indicating that the isolated platelets were highly purified and not contaminated by WBC. PM-NV was prepared via coating the purified platelet membranes on the surface of NV. Monodispersed NV was obtained with a particle size of 105 nm and zeta potential of −2.0 mV (Figure S2), characterized by the dynamic light scattering (DLS). After coating with PM, the resulting PM-NV appeared as a core-shell structure with an average diameter of 120.9 nm and zeta potential of −21.3 mV (Figure 2a). The increased size and decreased surface charge indicated the existence of PM on the surface of NV[4] (Figure 2b). Additionally, the successful coating of PM was further confirmed by colocalization of the FITC-labeled PM and rhodamine-labeled NV (Figure S3). To investigate the stability of PM-NV, the size changes of NV and PM-NV was monitored by the absorbance method.[18] The absorbance values measured at 560 nm and particle size suggested the superior serum stability of PM-NV when compared with NV (Figure 2c and Figure S4).

Figure 2.

Figure 2

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Characterization of PM-NV. a), The TEM image and hydrodynamic size distribution of PM-NV. Arrow indicates the existence of platelet membrane. The scale bar: 100 nm; inset: 50 nm. b), Changes in particle size and zeta potential of NV after coating with PM. Error bars indicate s.d. (n=3). c). In vitro stability of NV and PM-NV in the 100% fetal bovine serum. The absorbance at 560 nm is monitored. Error bars indicate s.d. (n=3). d), In vitro release of Dox from NV and PM-NV in PBS with different pH. Error bars indicate s.d. (n=3).

To characterize the pH-responsive release behavior of Dox from PM-NV, a nondegradable crosslinker, methylenebis-(acrylamide)[19] was applied to substitute glycerol dimethacrylate (GDA) to prepare nondegradable NV as a control. The in vitro release profile of Dox from Dox-NV and Dox-PM-NV was monitored at pH 5.4, 6.5 and 7.4, respectively (Figure 2d and Figure S5). Only about 23% of Dox was released from Dox-NV and Dox-PM-NV within 6 h and approximately 65% of Dox was released within 48 h at pH 7.4. In contrast, Dox-NV and Dox-PM-NV showed much higher cumulative Dox release at pH 5.4 than that of Dox-NV and Dox-PM-NV at pH 7.4. More than 40% of Dox was released in the first 6 h from both Dox-NV and Dox-PM-NV and about 88% of Dox was released within 48 h (Figure 2d). Furthermore, it is also noteworthy that the coating of PM could decline the release rate of Dox, suggesting that the PM could inhibit the burst release of drugs from NV. Collectively, it was demonstrated that the acidic environment accelerated the dissociation of PM-NV and subsequent release of encapsulated Dox.

To investigate the delivery efficacy of TRAIL and Dox by PM-NV, the human breast adenocarcinoma (MDA-MB-231) cells[20] were incubated with NV and PM-NV for 2 h, followed by the observation via confocal laser scanning microscopy (CLSM). An evident difference of distribution of TRAIL delivered by NV and PM-NV was recorded. As displayed in Figure 3a, the distribution of the rhodamine-labled PM-NV was located on the cellular membrane and cytoplasm. In contrast, all of the rhodamine-labeled non-coated NV was found in the cytoplasm, indicating directly decorated TRAIL on the surface of NV could be easily transported into the cells. The efficient delivery of TRAIL toward plasma membrane mediated by PM-NV could be attributed to the selective affinity of platelets and cancer cells. The P-Selectin protein overexpressed on the surface of platelets could specifically bind to the CD44, an upregulated receptors on the cancer cell membrane. Collectively, the selective binding interaction between platelets and cancer cells led to an efficient extracellular delivery of TRAIL, subsequently triggering the following extrinsic apoptosis pathway.

Figure 3.

Figure 3

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In vitro site-specific delivery of TRAIL and Dox by PM-NV. a), Extracellular distribution of TRAIL-NV and TRAIL-PM-NV after 2 h incubation. White arrows indicate the location of rhodamine-labeled TRAIL-NV and TRAIL-PM-NV. Scale bar: 20 μm. b), The induced apoptosis of MDA-MB-231 cells treated with TRAIL-Dox-NV and TRAIL-Dox-PM-NV after incubation for 12 h using the APO-BrdU TUNEL assay. Green fluorescence indicates Alexa Fluor 488-stained nick end label DNA fragment, and red fluorescence indicates PI-stained nuclei. Scale bar: 100 μm. c), Intracellular delivery of Dox-PM-NV on MDA-MB-231 cells at different time observed by CLSM. The late endo-lysosomes were stained by LysoTracker Green, and the nuclei were stained by Hoechst 33342. Scale bar: 20 μm. d), Flow cytometry analysis of MDA-MB-231 cells after staining with Annexin V-FITC and PI. The cells were treated with TRAIL-Dox-NV, TRAIL-PM-NV, Dox-PM-NV and TRAIL-Dox-PM-NV at the TRAIL concentration of 20 ng/mL and Dox concentration of 200 ng/mL for 12 h. The cells incubated with drug-free DMEM were served as a control. e), In vitro cytotoxicity of TRAIL-Dox-NV, TRAIL-PM-NV, Dox-PM-NV and TRAIL-Dox-PM-NV after incubation for 24 h. Error bars indicate s.d. (n=3).

Next, we evaluated the intracellular delivery of Dox by PM-NV using CLSM. PM-NV was demonstrated to be internalized by the MDA-MB-231 cells via the clathrin-dependent endocytosis with the involvement of lipid raft (Figure S6). As shown in Figure 3c, the majority of the internalized PM-NVs were found in the endo-lysosomes labeled with the green fluorescence, visualized by the overlaid yellow fluorescence for 1 h incubation. In contrast, a large number of endocytosed PM-NVs were localized in the nuclei stained with blue fluorescence, evidenced by the overlaid magenta fluorescence after 4 h incubation. The enhanced accumulation of Dox in the nuclei indicated the efficient nuclei delivery of Dox that was contributed by the dissociation of GDA matrix in the acidic endo-lysosome. Furthermore, the uptake efficiency of Dox-PM-NV on Raw264.7 cells was significantly lower than that of Dox-NV (Figure S7), indicating the “self-recognized” proteins presented on PM could inhibit the uptake by macrophage cells, which is the main reason for quick clearance of drug delivery systems (DDS) in vivo.

We further applied the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay[12a] and the Annexin-V/PI double staining assay[21] to substantiate the synergetic apoptosis inducing capability of PM-NV. A stronger green fluorescence was visualized by TRAIL-Dox-PM-NV compared with TRAIL-Dox-NV, indicated the increased apoptotic DNA fragment (Figure 3b). In addition, the quantitative flow cytometry results showed the rate of early and late apoptosis of MDA-MB-231 cells after incubation with TRAIL-Dox-PM-NV for 12 h were 31.8% and 12.3%, respectively, which was significantly higher than 22.0% and 0.4% of TRAIL-Dox-NV, 17.4% and 4.4% of Dox-PM-NV, and 12.6% and 5.6% of TRAIL-PM-NV (Figure 3d).

The in vitro cytotoxicity of TRAIL-Dox-PM-NV against MDA-MB-231 cells was evaluated by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay.[11b, 12a] TRAIL-Dox-PM-NV exhibited significantly enhanced cytotoxicity compared with TRAIL-Dox- NV, TRAIL-PM-NV and Dox-PM-NV at all studied TRAIL and Dox concentrations (Figure 3e). Notably, the TRAIL-Dox-PM-NV exhibited the increased cytotoxicity with decreased IC50 values of 19.3 ng/mL (TRAIL concentration) and 193 ng/mL (Dox concentration), 2.2-fold lower than that of TRAIL-Dox-NV, 2.6-fold lower than that of Dox-PM-NV, and 7.8-fold than that of TRAIL-PM-NV. Collectively, these results validated that the sequential and sit-specific delivery of TRAIL and Dox enabled by PM-NV effectively initiated a synergistic induction of apoptosis through the combination efficacy of TRAIL and Dox.

To evaluate the tumor targeting capability of PM-NV, Cy5.5-labeled TRAIL-Dox-PM-NV was administrated intravenously into the MDA-MB-231 tumor-bearing nude mice via tail vein. PM-NV exhibited strong fluorescence signal at the tumor site at 6 h post-injection (Figure 4a), validating the notable targeting ability of PM-NV, which can selectively bind to the overexpressed CD44 receptors on the surface of MDA-MB-231 cells. As time extended, elevated fluorescence intensity was found at the tumor site of mice treated with PM-NV when compared with that treated with NV at 12 h, 24 h and 48 h (Figure 4a), indicating superiority of the active targeting capability mediated by platelet membranes than EPR effect. Additionally, a prolonged retention time at tumor site was achieved by PM-NV at 48 h post injection. After 48 h imaging, the tumor and normal tissues were taken out for ex vivo imaging. The fluorescence intensity of PM-NV at tumor site was much higher than that of NV and other organs (Figure 4b). The results were confirmed by the quantitative region-of-interest (ROI) analysis, which showed 1.9-fold higher fluorescence intensity than that of NV, as well as 3.0-fold and 4.5-fold higher than that of liver and kidney (Figure 4c). Additionally, the enhanced accumulation of PM-NV at tumor site was further validated by the distribution of Dox, which displayed the increased red fluorescence signal of Dox when compared with NV (Figure 4d). The pharmacokinetics of PM-NV administered intravenously was evaluated by quantitatively monitoring the Dox concentration in the blood plasma. The elimination half-life (t1/2) and the AUC (area under the curve) were significantly higher than those of the NV, suggesting the capability of PM-NV to maintain a prolonged circulation time (Figure S8). These in vivo findings further confirmed that the NV coated with the platelet membranes that contained “self-recognized” proteins[2a] could inhibit macrophage uptake, which was in good accordance with in vitro macrophage uptake results.

Figure 4.

Figure 4

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In vivo targeting ability of PM-NV. a), In vivo fluorescence imaging of the MDA-MB-231 tumor-bearing nude mice at 6, 12, 24 h and 48 h after intravenous injection of Cy5.5-labeled TRAIL-Dox-NV (i) and Cy5.5-labeled TRAIL-Dox-PM-NV (ii) at Cy5.5 dose of 20 nmol/kg. Arrows indicate the sites of tumors. b), Ex vivo fluorescence imaging of the excised tumors and normal tissues at 48 h post injection. i, Cy5.5-TRAIL-Dox-NV; ii, Cy5.5-TRAIL-Dox-PM-NV. From top to bottom, 1: tumor; 2: kidney; 3: lung; 4: spleen; 5: liver; 6: heart. c), Region-of-interest analysis of fluorescent intensities from the tumors and normal tissues. Error bars indicate s.d. (n=3). *P<0.05 (two-tailed Student's t-test). d), In vivo fluorescence images of tumors treated with TRAIL-Dox-NV and TRAIL-Dox-PM-NV. Scale bar: 100 μm.

The in vivo antitumor efficacy was then assessed using the MDA-MB-231 tumor-bearing nude mice. The growth of the tumor was significantly inhibited after the treatment with different TRAIL/Dox formulations, including TRAIL-Dox-NV, TRAIL-PM-NV, Dox-PM-NV and TRAIL-Dox-PM-NV, compared with the saline control group (Figure 5a, b). The tumor treated with TRAIL-Dox-PM-NV showed the remarkably smaller volume compared with TRAIL-PM-NV and Dox-PM-NV, which indicated the synergetic antitumor efficacy enabled by PM-NV with the combination of TRAIL and Dox. Additionally, the strongest antitumor effect achieved by TRAIL-Dox-PM-NV suggested the sequential and site-specific delivery of TRAIL and Dox to their most active destinations could strengthen the synergetic antitumor efficacy. Meanwhile, the body weight of mice receiving different drug formulations remained stable during the treatment (Figure S9). The histologic images of the tumor section stained by the hematoxylin and eosin (H&E) showed a massive cancer cell remission after treated with TRAIL-Dox-PM-NV (Figure S10). In addition, the fluorescence images obtained using the in situ TUNEL assay presented the highest level of cell apoptosis in the tumor collected from the mice treated with TRAIL-Dox-PM-NV (Figure S10). No obvious pathological abnormalities in the heart, such as cardiomyopathy, the main toxic effect in Dox cancer treatment, were found for TRAIL-Dox-NV, Dox-PM-NV and TRAIL-Dox-PM-NV (Figure S11). Furthermore, no obvious pathological abnormalities were observed on normal organs (Figure S12).

Figure 5.

Figure 5

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In vivo antitumor efficacy evaluation and elimination of circulating tumor cells (CTCs). a), Representative images of the MDA-MB-231 tumors after treatment with different TRAIL/Dox formulations at day 16 (from top to bottom, 1: saline, 2: TRAIL-Dox-NV, 3: TRAIL-PM-NV, 4: Dox-PM-NV, 5: TRAIL-Dox-PM-NV) at TRAIL dose of 1 mg/kg and Dox dose of 2 mg/kg. b), The MDA-MB-231 tumor growth curves after intravenous injection of different TRAIL/Dox formulations. Error bars indicate s.d. (n=5). *P<0.05 (two-tailed Student's t-test). c), Representative images of the lung tissues 8 weeks post intravenous injection with MDA-MB-231 cells and different TRAIL/Dox formulations. Red arrows indicate the visible metastatic nodules. d), Histological observation of the lung tissues after treatment. The lung sections were stained with hematoxylin and eosin. Black arrows indicate the tumor cells. Scale bar: 200 μm. e), Quantification of visible metastatic nodules. i: Saline; ii: TRAIL-Dox-NV; iii: TRAIL-Dox-PM-NV. Error bars indicate s.d. (n=3). ***P<0.001 (two-tailed Student's t-test).

To further assess the potency of this PM-coated combinational drug delivery platform, we investigate the in vivo capability of elimination of CTCs by injecting nude mice with MDA-MB-231 cells (1 × 106 cells/100 μL saline). These circulating tumor cells could spread to various organs particularly into lungs.[22] As displayed in Figure 5c, the mice treated with saline exhibited remarkable lung metastasis, which was confirmed by the H&E staining (Figure 5d). However, the mice treated with TRAIL-Dox-NV showed slightly decrease in lung metastasis with no significant difference compared with the saline group. In sharp contrast, the significantly reduced metastatic nodules were found at the lung of mice treated with TRAIL-Dox-PM-NV when compared to the lung of mice treated with saline and TRAIL-Dox-NV (Figure 5e), suggesting the efficient elimination of CTCs in the blood stream which was attributed to the selective capture by P-Selectin on PM and subsequent activation of apoptosis by the binding of TRAIL and death receptors and accumulation of Dox into the nuclei.[14a]

In conclusion, we have developed a platelet membrane-coated nano-formulation for sequential and site-specific delivery of TRAIL and Dox. By taking advantage of the specific affinity between platelets and cancer cells, the PM-NV can efficiently deliver TRAIL toward cancel cell membrane to activate the extrinsic apoptosis signaling pathway. Equipped with an acid-responsive encapsulation matrix, the PM-NV can be digested after endocytosis and enhanced the Dox accumulation at the nuclei for activation of the intrinsic apoptosis pathway. The promising synergetic antitumor efficacy was achieved by TRAIL-Dox-PM-NV. Because of the serum stability, targeting specificity, and ease of generation, this PM-NV could also deliver other proteins that act on the tumor cellular membrane, such as cetuximab and trastuzumab to achieve a synergetic antitumor efficacy, with combination of other intracellular therapeutics. Importantly, since the metastatic cancer cells need platelets to aggregate around them to help cancer cells survive in blood and spread to new tissues,[8a] this PM-NV could further be adapted to identify and eliminate tumor cells that have the metastasis potentiality. Finally, since platelets also play a key role in several physiologic and pathologic processes such as hemostasis and thrombosis by forming the plugs that seal injured vessels and arrest bleeding,[23] this platform also holds promise in treating vascular disease.

Supplementary Material

Supporting Information

Acknowledgements

This work was supported by the grants from NC TraCS, NIH's Clinical and Translational Science Awards (CTSA, NIH grant 1UL1TR001111) at UNC-CH. We greatly thank Dr. Elizabeth Loboa, Dr. Michael Gamcsik and Dr. Glenn Walker for providing experimental facilities. We greatly thank Dr. Seulki Lee and Dr. Tae Hyung Kim at Johns Hopkins School of Medicine for offering us Escherichia coli containing the pET23dw-His-ILZ-hTRAIL vector as a gift. We acknowledge the use of the Analytical Instrumentation Facility (AIF) at NC State, which is supported by the State of North Carolina and the National Science Foundation (NSF).

Footnotes

Supporting Information

Supporting Information is available from the Wiley Online Library or from the author.

Contributor Information

Quanyin Hu, Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27695, USA; Division of Molecular Pharmaceutics and Center for Nanotechnology in Drug Delivery, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.

Wujin Sun, Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27695, USA; Division of Molecular Pharmaceutics and Center for Nanotechnology in Drug Delivery, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.

Chengen Qian, Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27695, USA; Division of Molecular Pharmaceutics and Center for Nanotechnology in Drug Delivery, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.

Dr. Chao Wang, Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27695, USA Division of Molecular Pharmaceutics and Center for Nanotechnology in Drug Delivery, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.

Hunter N. Bomba, Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27695, USA Division of Molecular Pharmaceutics and Center for Nanotechnology in Drug Delivery, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA.

Prof. Dr. Zhen Gu, Joint Department of Biomedical Engineering, University of North Carolina at Chapel Hill and North Carolina State University, Raleigh, NC 27695, USA; Division of Molecular Pharmaceutics and Center for Nanotechnology in Drug Delivery, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599, USA; Department of Medicine, University of North Carolina School of Medicine, Chapel Hill, NC 27599, USA.

References

  • 1.a Sarikaya M, Tamerler C, Jen AK-Y, Schulten K, Baneyx F. Nature materials. 2003;2:577–585. doi: 10.1038/nmat964. [DOI] [PubMed] [Google Scholar]; b Yoo J-W, Irvine DJ, Discher DE, Mitragotri S. Nature reviews Drug discovery. 2011;10:521–535. doi: 10.1038/nrd3499. [DOI] [PubMed] [Google Scholar]; c Gao W, Fang RH, Thamphiwatana S, Luk BT, Li J, Angsantikul P, Zhang Q, Hu C-MJ, Zhang L. Nano letters. 2015;15:1403–1409. doi: 10.1021/nl504798g. [DOI] [PMC free article] [PubMed] [Google Scholar]; d Wang Q, Cheng H, Peng H, Zhou H, Li PY, Langer R. Advanced drug delivery reviews. 2014 doi: 10.1016/j.addr.2014.12.003. [DOI] [PubMed] [Google Scholar]; e Wang C, Sun X, Cheng L, Yin S, Yang G, Li Y, Liu Z. Advanced Materials. 2014;26:4794–4802. doi: 10.1002/adma.201400158. [DOI] [PubMed] [Google Scholar]
  • 2.a Hu C-MJ, Zhang L, Aryal S, Cheung C, Fang RH, Zhang L. Proceedings of the National Academy of Sciences. 2011;108:10980–10985. doi: 10.1073/pnas.1106634108. [DOI] [PMC free article] [PubMed] [Google Scholar]; b Hu C-MJ, Fang RH, Copp J, Luk BT, Zhang L. Nature nanotechnology. 2013;8:336–340. doi: 10.1038/nnano.2013.54. [DOI] [PMC free article] [PubMed] [Google Scholar]; c Hu C-MJ, Fang RH, Luk BT, Zhang L. Nature nanotechnology. 2013;8:933–938. doi: 10.1038/nnano.2013.254. [DOI] [PMC free article] [PubMed] [Google Scholar]; d Copp JA, Fang RH, Luk BT, Hu C-MJ, Gao W, Zhang K, Zhang L. Proceedings of the National Academy of Sciences. 2014;111:13481–13486. doi: 10.1073/pnas.1412420111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3.Parodi A, Quattrocchi N, van de Ven AL, Chiappini C, Evangelopoulos M, Martinez JO, Brown BS, Khaled SZ, Yazdi IK, Enzo MV. Nature nanotechnology. 2013;8:61. doi: 10.1038/nnano.2012.212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Fang RH, Hu C-MJ, Luk BT, Gao W, Copp JA, Tai Y, O'Connor DE, Zhang L. Nano letters. 2014;14:2181–2188. doi: 10.1021/nl500618u. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5.a Doshi N, Orje JN, Molins B, Smith JW, Mitragotri S, Ruggeri ZM. Advanced Materials. 2012;24:3864–3869. doi: 10.1002/adma.201200607. [DOI] [PMC free article] [PubMed] [Google Scholar]; b Liu X, Zhang F, Wang Q, Gao J, Meng J, Wang S, Yang Z, Jiang L. Small. 2014;10:4677–4683. doi: 10.1002/smll.201401530. [DOI] [PubMed] [Google Scholar]; c Gay LJ, Felding-Habermann B. Nature Reviews Cancer. 2011;11:123–134. doi: 10.1038/nrc3004. [DOI] [PMC free article] [PubMed] [Google Scholar]; d Nguyen DX, Bos PD, Massagué J. Nature Reviews Cancer. 2009;9:274–284. doi: 10.1038/nrc2622. [DOI] [PubMed] [Google Scholar]
  • 6.a Nash G, Turner L, Scully M, Kakkar A. The lancet oncology. 2002;3:425–430. doi: 10.1016/s1470-2045(02)00789-1. [DOI] [PubMed] [Google Scholar]; b Labelle M, Begum S, Hynes RO. Cancer cell. 2011;20:576–590. doi: 10.1016/j.ccr.2011.09.009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7.Chaffer CL, Weinberg RA. Science. 2011;331:1559–1564. doi: 10.1126/science.1203543. [DOI] [PubMed] [Google Scholar]
  • 8.a Borsig L, Wong R, Feramisco J, Nadeau DR, Varki NM, Varki A. Proceedings of the National Academy of Sciences. 2001;98:3352–3357. doi: 10.1073/pnas.061615598. [DOI] [PMC free article] [PubMed] [Google Scholar]; b Stone J, Wagner D. Journal of Clinical Investigation. 1993;92:804. doi: 10.1172/JCI116654. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.a Jurasz P, Alonso-Escolano D, Radomski MW. British journal of pharmacology. 2004;143:819–826. doi: 10.1038/sj.bjp.0706013. [DOI] [PMC free article] [PubMed] [Google Scholar]; b Labelle M, Begum S, Hynes RO. Proceedings of the National Academy of Sciences. 2014;111:E3053–E3061. doi: 10.1073/pnas.1411082111. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.George D. Seminars in oncology. Vol. 28. Elsevier; 2001. pp. 27–33. [PubMed] [Google Scholar]
  • 11.a Jiang T, Mo R, Bellotti A, Zhou J, Gu Z. Advanced Functional Materials. 2014;24:2295–2304. [Google Scholar]; b Jiang T, Sun W, Zhu Q, Burns NA, Khan SA, Mo R, Gu Z. Advanced Materials. 2015;27:1021–1028. doi: 10.1002/adma.201404498. [DOI] [PMC free article] [PubMed] [Google Scholar]; c Pacardo DB, Ligler FS, Gu Z. Nanoscale. 2015;7:3381–3391. doi: 10.1039/c4nr07677j. [DOI] [PubMed] [Google Scholar]
  • 12.a Mo R, Jiang T, DiSanto R, Tai W, Gu Z. Nature communications. 2014:5. doi: 10.1038/ncomms4364. [DOI] [PubMed] [Google Scholar]; b Sun W, Jiang T, Lu Y, Reiff M, Mo R, Gu Z. Journal of the American Chemical Society. 2014;136:14722–14725. doi: 10.1021/ja5088024. [DOI] [PMC free article] [PubMed] [Google Scholar]; c Bagalkot V, Farokhzad OC, Langer R, Jon S. Angewandte Chemie International Edition. 2006;45:8149–8152. doi: 10.1002/anie.200602251. [DOI] [PubMed] [Google Scholar]; d Xiong M, Bao Y, Du X, Tan Z, Jiang Q, Wang H, Zhu Y, Wang J. ACS nano. 2013;7:10636–10645. doi: 10.1021/nn403146t. [DOI] [PubMed] [Google Scholar]
  • 13.a Peer D, Karp JM, Hong S, Farokhzad OC, Margalit R, Langer R. Nature nanotechnology. 2007;2:751–760. doi: 10.1038/nnano.2007.387. [DOI] [PubMed] [Google Scholar]; b Farokhzad OC, Langer R. ACS nano. 2009;3:16–20. doi: 10.1021/nn900002m. [DOI] [PubMed] [Google Scholar]
  • 14.a Mitchell MJ, Wayne E, Rana K, Schaffer CB, King MR. Proceedings of the National Academy of Sciences. 2014;111:930–935. doi: 10.1073/pnas.1316312111. [DOI] [PMC free article] [PubMed] [Google Scholar]; b Cristofanilli M, Budd GT, Ellis MJ, Stopeck A, Matera J, Miller MC, Reuben JM, Doyle GV, Allard WJ, Terstappen LW. New England Journal of Medicine. 2004;351:781–791. doi: 10.1056/NEJMoa040766. [DOI] [PubMed] [Google Scholar]; c Chambers AF, Groom AC, MacDonald IC. Nature Reviews Cancer. 2002;2:563–572. doi: 10.1038/nrc865. [DOI] [PubMed] [Google Scholar]
  • 15.a Qureshi AH, Chaoji V, Maiguel D, Faridi MH, Barth CJ, Salem SM, Singhal M, Stoub D, Krastins B, Ogihara M. PLoS One. 2009;4:e7627. doi: 10.1371/journal.pone.0007627. [DOI] [PMC free article] [PubMed] [Google Scholar]; b Donovan LE, Dammer EB, Duong DM, Hanfelt JJ, Levey AI, Seyfried NT, Lah JJ. Alzheimers Res Ther. 2013;5 doi: 10.1186/alzrt186. 10.1186. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Boilard E, Nigrovic PA, Larabee K, Watts GF, Coblyn JS, Weinblatt ME, Massarotti EM, Remold-O'Donnell E, Farndale RW, Ware J. Science. 2010;327:580–583. doi: 10.1126/science.1181928. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17.Aicher A, Heeschen C, Mildner-Rihm C, Urbich C, Ihling C, Technau-Ihling K, Zeiher AM, Dimmeler S. Nature medicine. 2003;9:1370–1376. doi: 10.1038/nm948. [DOI] [PubMed] [Google Scholar]
  • 18.a Popielarski SR, Pun SH, Davis ME. Bioconjugate chemistry. 2005;16:1063–1070. doi: 10.1021/bc050113d. [DOI] [PubMed] [Google Scholar]; b Fang RH, Aryal S, Hu C-MJ, Zhang L. Langmuir. 2010;26:16958–16962. doi: 10.1021/la103576a. [DOI] [PubMed] [Google Scholar]
  • 19.a Cohen JA, Beaudette TT, Tseng WW, Bachelder EM, Mende I, Engleman EG, Fréchet JM. Bioconjugate chemistry. 2008;20:111–119. doi: 10.1021/bc800338n. [DOI] [PMC free article] [PubMed] [Google Scholar]; b Liu Y, Du J, Yan M, Lau MY, Hu J, Han H, Yang OO, Liang S, Wei W, Wang H. Nature nanotechnology. 2013;8:187–192. doi: 10.1038/nnano.2012.264. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20.Sheridan C, Kishimoto H, Fuchs RK, Mehrotra S, Bhat-Nakshatri P, Turner CH, Goulet R, Jr, Badve S, Nakshatri H. Breast Cancer Res. 2006;8:R59. doi: 10.1186/bcr1610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21.Hu Q, Gu G, Liu Z, Jiang M, Kang T, Miao D, Tu Y, Pang Z, Song Q, Yao L. Biomaterials. 2013;34:1135–1145. doi: 10.1016/j.biomaterials.2012.10.048. [DOI] [PubMed] [Google Scholar]
  • 22.Price JE, Polyzos A, Zhang RD, Daniels LM. Cancer research. 1990;50:717–721. [PubMed] [Google Scholar]
  • 23.a Ruggeri ZM. Nature medicine. 2002;8:1227–1234. doi: 10.1038/nm1102-1227. [DOI] [PubMed] [Google Scholar]; b Davì G, Patrono C. New England Journal of Medicine. 2007;357:2482–2494. doi: 10.1056/NEJMra071014. [DOI] [PubMed] [Google Scholar]

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Supporting Information
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