Tumor-Targeted and Clearable Human Protein ... - ACS Publications

Subscriber access provided by UNIV OF ARIZONA

Communication

Tumor-Targeted and Clearable Human Protein-based MRI Nanoprobes Yang Zhao, Jing Peng, Jingjin Li, Ling Huang, Jingyi Yang, Kai Huang, Hewen Li, Ning Jiang, Shaokuan Zheng, Xuening Zhang, Yuanjie Niu, and Gang Han Nano Lett., Just Accepted Manuscript • Publication Date (Web): 05 Jun 2017 Downloaded from http://pubs.acs.org on June 6, 2017

Just Accepted “Just Accepted” manuscripts have been peer-reviewed and accepted for publication. They are posted online prior to technical editing, formatting for publication and author proofing. The American Chemical Society provides “Just Accepted” as a free service to the research community to expedite the dissemination of scientific material as soon as possible after acceptance. “Just Accepted” manuscripts appear in full in PDF format accompanied by an HTML abstract. “Just Accepted” manuscripts have been fully peer reviewed, but should not be considered the official version of record. They are accessible to all readers and citable by the Digital Object Identifier (DOI®). “Just Accepted” is an optional service offered to authors. Therefore, the “Just Accepted” Web site may not include all articles that will be published in the journal. After a manuscript is technically edited and formatted, it will be removed from the “Just Accepted” Web site and published as an ASAP article. Note that technical editing may introduce minor changes to the manuscript text and/or graphics which could affect content, and all legal disclaimers and ethical guidelines that apply to the journal pertain. ACS cannot be held responsible for errors or consequences arising from the use of information contained in these “Just Accepted” manuscripts.

Nano Letters is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.

Page 1 of 19

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Nano Letters

1

Tumor-Targeted and Clearable Human Protein-

2

based MRI Nanoprobes

3

Yang Zhao1,2,3,§, Jing Peng1,§, Jingjin Li1,§, Ling Huang3, Jinyi Yang3,Kai Huang3, Hewen Li1,

4

Ning Jiang2, Shaokuan Zheng3, Xuening Zhang1,*, Yuanjie Niu2,*, Gang Han3,*

5

1. Department of Radiology, The Second Hospital of Tianjin Medical University, Tianjin

6

300211, China

7

2. Sex Hormone Research Center, Tianjin Institute of Urology, Tianjin 300211, China

8

3. Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 364

9

Plantation Street, LRB 806 Worcester MA 01605

10

* Corresponding authors: [email protected] (XZ), [email protected] (YN),

11

[email protected] (GH)

12

§ These authors contribute equally to this work.

13 14 15 16 17

ACS Paragon Plus Environment

1

Nano Letters

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 2 of 19

1

Table of Contents Graphic:

2

A tumor-targeting and clearable human protein-based MRI contrast nanoprobe was developed.

3

Compared to the conventionally used gadolinium chelates, the resultant nanoprobes exhibited

4

superior longitudinal relaxation as well as retain the natural tumor targeting ability and the

5

subsequent tumor retrieval and systematic body clearance functions of human transferrin

6

proteins. We believe that such nanoprobes offer a new important approach to design

7

biocompatible functional MRI contrast agents and possess great potential for clinical

8

translations.

9

10 11


2

Page 3 of 19

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Nano Letters

1

Abstract: Biocompatibility, targeting, and clearance are key challenges in the design of new

2

MRI contrast agents. Herein, we report on a tumor-targeting, gadolinium biomineralized human

3

transferrin (Tf) protein-based nanoparticle (Gd@Tf NP) for MRI use. As compared to the

4

conventionally used gadolinium chelates, the resultant Gd@Tf NPs possess outstanding chemical

5

stability and exhibited superior longitudinal relaxation. More importantly, our MR images show

6

that Gd@Tf indeed retained the natural tumor targeting ability and the subsequent tumor

7

retrieval biofunctions of Tf. Thus, such Tf protein-based MR NPs integrate T1 signal

8

amplification, precise tumor targeting, and systematic clearance capabilities. They offer a new

9

approach to design biocompatible multifunctional MRI contrast agents for a wide range of

10

clinical imaging and treatment applications.

11 12

Keywords: magnetic resonance imaging, magnetic nanoparticles, protein, tumor targeting,

13

clearance

14 15 16 17 18 19 20


3

Nano Letters

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 4 of 19

1

MRI is one of the most used non-invasive, versatile imaging modalities for the clinical detection,

2

staging, and monitoring of the treatment of tumors. This is the case due to its unique advantages,

3

including high spatial resolution, outstanding soft tissue contrast, and since it also causes no

4

radiation damage. Recently, the development of magnetic contrast agents (CAs) have improved

5

the inherent sensitivity of MRI, enabling this technique to visualize specific biological processes

6

at both cellular and molecular levels.1, 2 To date, the most frequently used magnetic CAs in

7

clinical settings are Gd-based chelates due to their signal-enhancing positive contrast ability and

8

negligible immunogenicity.3 However, such compounds often suffer from short life spans,

9

relatively low relaxivity, and the consequent need for high doses of intravenous administration,

10

as well as the problematic gadolinium retention within vital organs.4-6 To overcome these

11

drawbacks, inorganic based Gd-containing nanoparticles have been explored in attempts to

12

enhance MR imaging quality such as gadolinium oxide, gadolinium fluoride, and gadolinium

13

phosphate. Such nanoparticles reveal the merits of large longitudinal relaxation, enhanced

14

sensitivity, prolonged circulating time and facile chemical modifications.2, 5 However, their long-

15

term stability, biocompatibility and clearance remain largely unclear, hindering their further

16

biomedical applications.7, 8 Thus, it is still a key challenge and highly desired to develop a

17

biocompatible tumor-targeting, as well as systemically clearable and more efficient Gd-based

18

nanoparticle MRI CAs.

19

Nowadays, proteins, as naturally occurring nanomaterials, hold great promise for clinical

20

applications.9 For example, albumin-bound paclitaxel nanoparticle Abraxane® has been

21

approved for clinic use in treating metastatic breast cancer.10 Similarly, the use of proteins as

22

scaffolds to encapsulate metal-based nanoparticulate CAs has been proven to be able to enhance

23

the relaxivity of metallic CAs and to simultaneously provide additional active sites for further


4

Page 5 of 19

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Nano Letters

1

multi-functionalizations.11-14 In particular, biomineralization provides a facile and effective

2

approach for the formation of new functional materials within the proteins. Certain nanoparticles

3

have been directly biomineralized in proteins and applied for tumor detection and therapy.14-19

4

Herein, we report on a transferrin (Tf) protein biomineralized Gd-based MRI CAs that, not only

5

exhibits superior paramagnetic properties compared to conventional Gd-based CAs, but also

6

maintains the normal physiological functions of the intact Tf protein (e.g., excellent

7

biocompatibility, negligible immunogenicity, outstanding tumor-targeting and body-clearable

8

abilities).

9

In human bodies, transferrin protein is an endogenous glycoprotein that plays an

10

important role in the metabolism of iron. In particular, Tf proteins act as iron ion shuttles in Tf

11

expressed cells. Upon binding to the transferrin receptor (TfR) on the cell surface, it can deliver

12

iron ion payloads into cells via receptor-mediated endocytosis. After iron being released into the

13

cytoplasm, iron-free Tf recycles back to the extracellular space. Since cancer cell proliferation

14

leads to a great need for iron, TfR is found to be overexpressed in many malignancies at levels

15

many times higher than those in normal cells, making TfR an essential target for specific

16

recognition of tumor cells.20-22 As a natural ligand of TfR, Tf has thus been widely used as an

17

additional adjuvant moiety to be conjugated to varied types of nanoparticles for the targeting of

18

tumors.23-27 Yet, the resultant nanoparticles are much larger in size than unmodified nanoparticles

19

and Tf alone, thus significantly altering physiological functions.

20

In this study, we utilize endogenous human Tf as the biotic template to fabricate Tf-based

21

gadolinium nanoparticles via the biomineralization processes. In vitro and in vivo results

22

demonstrate that the prepared Gd@Tf NPs preserve the favorable intrinsic characteristics of Tf

23

(Figure 1), such as biocompatibility, precise tumor-targeting, and efficient body clearance


5

Nano Letters

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

1

through the hepatobiliary system, in addition to its superior chemical and physical properties,

2

including high T1 relaxivity and excellent stability (Figure 2 and S6). To the best of our

3

knowledge, this is the first time that human protein-based MRI nanoparticles with tumor

4

targeting and outstanding clearance. In addition, these NPs are flexible for future functional

5

modifications. As a consequence, the designed Gd@Tf NPs hold a great potential for a wide

6

range of future clinical applications as a promising and advanced MRI contrast agent.

7

The Gd@Tf NPs were prepared by mimicking the natural biomineralization process. 15, 18 In

8

brief, 2.7mg of Gd(NO3)3 and 30mg of Tf were dissolved in 1.2 mL of pure water at 37° C and

9

stirred for 5 min. Then 0.06mL of NaOH (1M) was added to the mixture and the subsequent

Page 6 of 19

10

solution was stirred for 12 hours at 37° C. After purification, the final Gd@Tf NPs were obtained

11

as a transparent solution (Figure S1); and the 280nm peak of UV-VIS absorbance spectroscopy

12

confirmed the presence of protein components in Gd@Tf NPs (Figure S2). The average

13

hydrodynamic diameter of Gd@Tf NPs was determined as ≈ 9 nm by dynamic light scattering

14

(DLS; Figure 2A). The zeta-potential of Tf showed no obvious change (from –8.73 mV to –

15

6.27 mV) after the reaction. Moreover, energy dispersive X-ray (EDX) spectroscopy studies

16

(Figure S3) suggests the Gd2O3 within the Tf-protein template. Transmission electron

17

microscopy (TEM; Figure S4) images revealed the encapsulated Gd2O3 is ~2-4 nm in diameter.

18

In addition, the morphology of Gd@Tf NPs was further described using atomic force

19

microscopy (AFM; Figure S5), showing that the particles had an average thickness of 4.95±0.17

20

nm.

21 22

In order to demonstrate the paramagnetic capacity of Gd@Tf NPs, the T1 relaxivity was determined by the slope of the 1/T1 (R1) plot versus the Gd(III) ion concentration (Figure 2B).


6

Page 7 of 19

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Nano Letters

1

Our Gd@Tf NPs (17.42 mM−1s−1, Figure 2B) were found to have much higher T1 relaxivity than

2

that of commercial MRI contrast agent Magnevist (3-5 mM−1s−1), potentially attributed to the

3

enhancement effect of proteins on the relaxivity of metal ions.11, 28 Gd@Tf NPs solutions also

4

exhibited an obvious dose-dependent brightening effect on the T1-weighted MR images;

5

whereas Tf solutions and deionized water triggered no changes in signal intensity at various

6

concentrations (Figure S6). Following storage at 4° C for 3 months, excellent dispersion

7

stabilities were observed in the prepared Gd@Tf NPs with respect to their colloidal and magnetic

8

properties, benefiting the end use and storage (Figure S7).

9

The above results demonstrate that prepared Gd@Tf NPs are able to be tested as T1 MRI

10

contrast agent candidates due to their ultra-small size of Gd2O3 within the protein, high T1

11

enhancement effects and good stability. To further test their potential for biomedical

12

applications, the cytotoxicity of Gd@Tf NPs was then evaluated by methyl thiazolyltetrazolium

13

(MTT) assay. We found that PC-3 cell viabilities were not influenced by Gd@Tf NPs, even at

14

high doses of Gd of up to 98 µM (Figure S8). These results indicate that the template of Tf offers

15

great biocompatibility to Gd-based Tf NPs which is essential with respect to applications of

16

Gd@Tf NPs.

17

Next, the cellular uptake of Gd@Tf NPs was investigated by T1-weighted MRI. The

18

treatment of Gd@Tf NPs resulted in a remarkable increase in T1 signal intensity within PC-3

19

cells (985.82±114.47) relative to cells treated with PBS (310.11±33.95), indicating that Gd@Tf

20

NPs were able to be efficiently internalized by cancer cells (Figure 2C, S9, and S10).

21

Subsequently, like the Tf proteins, Gd@Tf NPs were found to be able to be shuttled out of the

22

treated cells, as their signal intensity in cells decreased over time (Figure S9). Competition

23

experiments were further performed to evaluate whether Tf of Gd@Tf NPs preserved their


7

Nano Letters

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 8 of 19

1

biological functions to specifically bind to TfRs, and thus mediate endocytosis of Gd@Tf NPs

2

through Tf-TfR interactions. As shown in Figures 2C and S10, the signal intensity of PC-3 cells

3

that were treated by excess Tf followed by Gd@Tf NPs was 406.53±22.89, which was

4

significantly lower than those subjected only to Gd@Tf NPs (985.82±114.47), revealing that the

5

pre-treatment of Tf protein only led to a significant decrease in signal intensity on T1 weighted

6

imaging. This result indicated that Tf protein indeed blocked binding sites for Gd@Tf NPs and

7

inhibit endocytosis of Gd@Tf NPs into tumor cells. As a result, the cellular uptake of Gd@Tf

8

NPs was mainly dependent on active targeting mechanisms mediated by the Tf-TfR delivery

9

system; and the biomineralization process did not interfere with the natural cell targeting and

10

shuttle functions of Tf in Gd@Tf NPs.

11

To further evaluate the MR performance of Gd@Tf NPs in vivo, a series of in-vivo MRI

12

was carried out. After tail-vein injection of Gd@Tf NPs, significantly enhanced signals were

13

observed in the tumor regions when compared to the pre-injection levels (Figure 3). The T1

14

signal intensity in the tumors peaked at 3 h post injection with a 3-fold increase in signal

15

intensity relative to the baseline (Figure S11). Such high contrast enhancement makes tumors

16

clearly distinguishable from the surrounding tissues. We believe that the enhanced permeability

17

and retention (EPR) effect is involved in the tumor-targeting accumulation of Gd@Tf NPs in

18

addition to the active targeting of Tfs, with the latter playing the predominant role in the tumor

19

cell targeting. In particular, our Gd@Tf NPs with hydrodynamic sizes of ~9 nm, can accumulate

20

in the tumor microenvironment via the EPR effect. After Gd@Tf NPs enter the tumor

21

microenvironment, they can then be internalized into tumor cells via active targeting with Tf

22

receptors that are over-expressed on tumor cells. Similar to the process of Fe3+ transport by Tf in

23

physiological reactions, Gd@Tf NPs were rapidly taken up into, and efficiently excreted from,


8

Page 9 of 19

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Nano Letters

1

tumor cells. Thus, after the peak time, MR signal intensity in tumor tissues gradually decreased

2

from their central regions and finally returned to the baseline level.

3

In order to ensure biosafety for further clinical translation, it is important to evaluate the

4

bio-clearance and the metabolism of magnetic nanoparticles after systematic administration. The

5

dynamic MR signal transformation of metabolic organs was carefully monitored to provide a

6

real-time visualization of the Gd@Tf metabolism. At baseline, the gall bladder and liver showed

7

hypointensity like water and isointensity as soft tissue on T1WI, respectively. 30min after

8

injection, the T1 signal of liver increased, while little change was observed in the signal intensity

9

of the gall bladder. At 4 h post-injection, the gall bladder began to demonstrate a heterogeneous

10

hyperintensity. Subsequently, its T1 signal gradually increased, and a very bright gall bladder

11

was then detectable on T1WI from 8 h to 15h, which made it quite different from the images

12

before 0.5h (Figure 4). The T1 signal of intestines was subsequently investigated. No obvious

13

change was found in intestinal during the first hour post-injection. At 4 h, part of the intestine

14

began to brighten and then the hyperintense area in the enteric tract gradually expanded, and

15

clearly outlined the entire intestinal shape at 15h post-injection. Furthermore, we collected all of

16

the mice’s feces during the first 2 days after injection. The T1-value analysis revealed the

17

obvious shorter longitudinal relaxation time of the feces in the Gd@Tf NPs-treated group than

18

that of the PBS-treated group, demonstrating that Gd@Tf being excreted out of the body with

19

animal waste (Figure S12). Finally, the MRI findings after 14 days showed that the T1 signal of

20

intensity had nearly come back to the baseline level (Figure S13). The above results suggested

21

the metabolic process of the prepared Gd@Tf NPs is similar to intact Tf. They were cleared by

22

the hepatobiliary systems and the following elimination with normal intestinal tract movement.

23

On the other hand, unlike nanoparticles smaller than 6 nm in diameter (e.g., Gd-chelates and Au


9

Nano Letters

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 10 of 19

1

nanoclusters),29, 30 no significant changes of MR signal were observed in kidneys. We believe that

2

the phenomenon is likely due to the fact that the hydrodynamic diameter of Gd@Tf NPs (≈9

3

nm) exceeds the size threshold of glomerular filtration membranes (4.5-5 nm) which preclude

4

their clearance via the renal system. 7, 31 More specifically, the Tf protein template in Gd@Tf

5

NPs retains its original biological functions. Physiologically, such natural proteins can only

6

undergo hepatic clearance rather than renal clearance, as they possess anionic charges and

7

molecular sizes beyond the filtration-size threshold. Therefore, our Gd@ Tf NPs still inherit the

8

metabolic pathway of hepatic clearance from the intact Tfs.

9

In order to investigate the toxicity of Gd@Tf in vivo, the body weight and histological

10

changes were also evaluated. Results showed no obvious body-weight loss in 2 weeks post

11

injection (Figure S14) or morphological changes in the organs studied in relation to the treated

12

mice (Figure S15). The blood examinations also revealed that the essential hematological

13

parameters were normal after treatment with Gd@Tf NPs for 14d (Table S1), demonstrating

14

their negligible immunogenicity. In addition, Gd content in major organs was determined by

15

ICP-MS analysis; and no obvious Gd was found to be retained in the vital organs after

16

completion of the main metabolic processes (Figure S16). Consequently, the above results

17

clearly confirmed the low toxicity of Gd@Tf NPs.

18

In summary, we have successfully synthesized tumor targeting and systematic clearable

19

gadolinium-Tf NPs by means of a straightforward, one-step, environmentally benign and

20

reproducible biominerazation method. More significantly, the Gd@Tf NPs retained the inherent

21

biological functions of Tf protein (i.e., biocompatibility, tumor targeting and systematic clearable

22

abilities) and the high paramagnetic property of Gd within a single nanoplatform. The MR


10

Page 11 of 19

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Nano Letters

1

Gd@Tf NPs have also demonstrated unique advantages over conventional nanoparticles, such as

2

having an ultra-small size and enhanced T1 MR signal amplification. More specifically, these

3

properties resulted in high-performing Gd@Tf NPs that showed selective MRI and precise tumor

4

localization in vivo. In addition, the protein-based MRI contrast agents were efficiently cleared

5

by the hepatobiliary system. These results show that Gd@Tf NPs are promising for clinical

6

implementation for activated and sensitive MR detection of early detection of tumor malignancy.

7

Furthermore, this preserved tumor shuttle-like feature of Tf in such nanoparticles would also

8

enable the prepared NPs to be applied as an MRI-imaging guided delivery vehicle for a wide

9

range of theranostic applications.


11

Nano Letters

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

1

Page 12 of 19

Figures

2 3

Figure 1. The Gd nanoparticles biomineralized in the template of Tf proteins. After tail vein

4

injection, Gd@Tf NPs were accumulated in the tumor areas and finally eliminated from the body

5

via the hepatobiliary system, demonstrating that Gd@Tf NPs are tumor targeting and

6

metabolically clearable.


12

Page 13 of 19

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Nano Letters

1

2 3

Figure 2. (a) Hydrodynamic size distributions and (b) r1 relaxivity curve for Gd@Tf NP. (c) In-

4

vitro T1-weighted MR images of PC-3 cell suspensions after treatment of PBS, Gd@Tf NPs, and

5

Tf+Gd@Tf NPs. The signal intensity of PC-3 cells that were treated by excess Tf followed by

6

Gd@Tf NPs were found to be significantly lower than those subjected only to Gd@Tf NPs.


13

Nano Letters

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 14 of 19

1

2 3

Figure 3. In-vivo T1-weighted MR images of PC-3 tumors before and after the intravenous

4

injection of Gd@Tf NPs. The significant signal enhancement was observed in the tumors (red

5

arrows) and reached its maximum point at ≈ 3 h post injection. After 3 h the MR signal

6

gradually descended from the central portion of the tumor region.


14

Page 15 of 19

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Nano Letters

1

2 3

Figure 4. In-vivo T1-weighted MR images of the gall bladder section (red circles) and intestine

4

section (blue arrows) in PC-3 tumor-bearing mice after the intravenous injection of Gd@Tf NPs.


15

Nano Letters

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Page 16 of 19

1

ASSOCIATED CONTENT

2

Supporting Information.

3

The Supporting Information is available free of charge on the ACS Publications website,

4

including Experimental Section, Figures S1-S15 and Table S1.

5

AUTHOR INFORMATION

6

Corresponding Author

7

*E-mail: [email protected] (XZ), [email protected] (YN), [email protected] (GH)

8

Author Contributions

9

YZ, JP and JL contributed equally to this paper. All authors have given approval to the final

10

version of the manuscript.

11

Notes

12

The authors declare no competing financial interest.

13

ACKNOWLEDGMENT

14

We thank Yuchen Lu (State Key Laboratory of Medicinal Chemical Biology at Nankai

15

University) for his assistance with in vivo biodistribution assays. This work was supported by the

16

National Institutes of Health R01MH103133 (G.H.), and the Human Frontier Science RGY-

17

0090/2014( G.H.) as well as the National Natural Science Foundation of China (Grants

18

81472682, 81572538, and 30700834). This work was also supported by the China Scholarship

19

Council (CSC) to Y.Z.

20

ABBREBIATIONS


16

Page 17 of 19

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

Nano Letters

1

MRI, magnetic resonance imaging; Tf, transferrin; NP, nanoparticles; CAs, contrast agents; TfR,

2

transferrin receptor; UV-VIS, ultraviolet visible; DLS, dynamic light scattering; EDX, energy

3

dispersive X-ray; TEM, transmission electron microscopy; MTT, methyl thiazolyltetrazolium.

4

References

5

1.

Sun, C.; Lee, J. S.; Zhang, M. Adv. Drug Delivery Rev. 2008, 60, 1252-1265.

6

2.

Zhang, L.; Liu, R.; Peng, H.; Li, P.; Xu, Z.; Whittaker, A. K. Nanoscale 2016, 8, 10491-

7

10510.

8

3.

9

2352.

10

4.

Na, H. B.; Hyeon, T. J. Mater. Chem. 2009, 19, 6267.

11

5.

McDonald, R. J.; McDonald, J. S.; Kallmes, D. F.; Jentoft, M. E.; Murray, D. L.; Thielen,

12

Caravan, P.; Ellison, J. J.; McMurry, T. J.; Lauffer, R. B. Chem Rev. 1999, 99, 2293-

K. R.; Williamson, E. E.; Eckel, L. J. Radiology 2015, 275, 772-782.

13

6.

14

457.

15

7.

Longmire, M.; Choyke, P. L.; Kobayashi, H. Nanomedicine (Lond). 2008, 3, 703-717.

16

8.

Chen, N.; Wang, H.; Huang, Q.; Li, J.; Yan, J. Small 2014, 10, 3603-3611.

17

9.

Kratz, F. J Control Release 2008, 132, 171-183.

18

10. Gradishar, W. J.; Tjulandin, S.; Davidson, N.; Shaw, H.; Desai, N.; Bhar, P.; Hawkins,

19

Tu, C.; Louie, A. Y. Wiley Interdiscip. Rev.: Nanomed. Nanobiotechnol. 2012, 4, 448-

M.; O'Shaughnessy, J. J Clin Oncol 2005, 23, 7794-7803.


17

Nano Letters

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

1

11. Koylu, M. Z.; Asubay, S.; Yilmaz, A. Molecules 2009, 14, 1537-1545.

2

12. Zhang, B.; Jin, H.; Li, Y.; Chen, B.; Liu, S.; Shi, D. J. Mater. Chem. 2012, 22, 14494.

3

13. Xie, J.; Zheng, Y.; Ying, J. Y. J. Am. Chem. Soc. 2009, 131, 888-889.

4

14. Wang, Y.; Yan, X. P. Chem. Commun. (Camb). 2013, 49, 3324-3326.

5

15. Wang, Y.; Yang, T.; Ke, H.; Zhu, A.; Wang, Y.; Wang, J.; Shen, J.; Liu, G.; Chen, C.;

6 7 8 9 10

Page 18 of 19

Zhao, Y.; Chen, H. Advanced Materials 2015, 27, 3874-3882. 16. Zhang, C.; Fu, Y. Y.; Zhang, X.; Yu, C.; Zhao, Y.; Sun, S. K. Dalton Trans. 2015, 44, 13112-13118. 17. Zhao, Y.; Peng, J.; Niu, Y.; Zhang, X.; Jiang, N.; Jia, R.; Li, J.; Shang, Z.; Zhu, S.; Sun, L. RSC Adv. 2015, 5, 64076-64082.

11

18. Sun, S.; Dong, L.; Cao, Y.; Sun, H.; Yan, X. Anal. Chem. 2013, 85, 8436-8441.

12

19. Wang, Z.; Huang, P.; Jacobson, O.; Wang, Z.; Liu, Y.; Lin, L.; Lin, J.; Lu, N.; Zhang, H.;

13 14 15

Tian, R.; Niu, G.; Liu, G.; Chen, X. ACS Nano 2016, 10, 3453-3460. 20. Hamilton, T. A.; Wada, H. G.; Sussman, H. H. Proc. Natl. Acad. Sci. U S A 1979, 76, 6406-6410.

16

21. Trowbridge, I. S.; Omary, M. B. Proc Natl Acad Sci U S A 1981, 78, 3039-3043.

17

22. Faulk, W. P.; Hsi, B. L.; Stevens, P. J. Lancet 1980, 2, 390-392.

18

23. Tortorella, S.; Karagiannis, T. C. Curr. Drug Deliv. 2014, 11, 427-443.


18

Page 19 of 19

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60

1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16

Nano Letters

24. Chen, X.; Sun, H.; Hu, J.; Han, X.; Liu, H.; Hu, Y. Colloids. Surf. B. Biointerfaces 2017, 152, 77-84. 25. Li, S.; Amat, D.; Peng, Z.; Vanni, S.; Raskin, S.; De Angulo, G.; Othman, A. M.; Graham, R. M.; Leblanc, R. M. Nanoscale 2016, 8, 16662-16669. 26. Wang, K.; Zhang, Y.; Wang, J.; Yuan, A.; Sun, M.; Wu, J.; Hu, Y. Sci Rep. 2016, 6, 27421. 27. Kang, T.; Jiang, M.; Jiang, D.; Feng, X.; Yao, J.; Song, Q.; Chen, H.; Gao, X.; Chen, J. Mol. Pharm. 2015, 12, 2947-2461. 28. Park, J. Y.; Baek, M. J.; Choi, E. S.; Woo, S.; Kim, J. H.; Kim, T. J.; Jung, J. C.; Chae, K. S.; Chang, Y.; Lee, G. H. ACS Nano 2009, 3, 3663-3669. 29. Grenier, N.; Mendichovszky, I.; de Senneville, B. D.; Roujol, S.; Desbarats, P.; Pedersen, M.; Wells, K.; Frokiaer, J.; Gordon, I. Semin. Nucl. Med 2008, 38, 47-55. 30. Yu, M.; Zhou, J.; Du B; Ning, X.; Authement, C.; Gandee, L.; Kapur, P.; Hsieh, J. T.; Zheng, J. Angew Chem Int Ed Engl. 2016, 55, 2787-91. 31. Ohlson, M.; Sorensson, J.; Haraldsson, B. Am. J. Physiol. Renal Physiol. 2001, 280, F396-405.


19

Tumor-Targeted and Clearable Human Protein ... - ACS Publications

Recommend Documents