Sarcoma-Targeting Peptide-Decorated Polypeptide Nanogel Intracellularly Delivers Shikonin for Upregulated Osteosarcoma Necroptosis and Diminished Pulmonary Metastasis - PubMed (original) (raw)

. 2018 Feb 2;8(5):1361-1375.

doi: 10.7150/thno.18299. eCollection 2018.

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Sarcoma-Targeting Peptide-Decorated Polypeptide Nanogel Intracellularly Delivers Shikonin for Upregulated Osteosarcoma Necroptosis and Diminished Pulmonary Metastasis

Suoyuan Li et al. Theranostics. 2018.

Erratum in

Abstract

Purpose: Osteosarcoma is the most common primary bone cancer and is notorious for pulmonary metastasis, representing a major threat to pediatric patients. An effective drug targeting osteosarcoma and its lung metastasis is urgently needed.

Design: In this study, a sarcoma-targeting peptide-decorated disulfide-crosslinked polypeptide nanogel (STP-NG) was exploited for enhanced intracellular delivery of shikonin (SHK), an extract of a medicinal herb, to inhibit osteosarcoma progression with minimal systemic toxicity.

Results: The targeted, loaded nanogel, STP-NG/SHK, killed osteosarcoma cells by inducing RIP1- and RIP3-dependent necroptosis in vitro. Necroptosis is a novel cell death form that could be well adapted as an efficient antitumor strategy, the main obstacle of which is its high toxicity. After intravenous injection, STP-NG/SHK efficiently suppressed tumor growth and reduced pulmonary metastasis, offering greater tumor necrosis and higher RIP1 and RIP3 upregulation compared to free SHK or untargeted NG/SHK in vivo. Additionally, the treatment with NG/SHK or STP-NG/SHK showed minimal toxicity to normal organs, suggesting low systemic toxicity compared to free SHK.

Conclusion: The STP-guided intracellular drug delivery system using the necroptosis mechanism showed profound anti-osteosarcoma activity, especially eliminated lung metastasis in vivo. This drug formulation may have great potential for treatment of osteosarcoma.

Keywords: Intracellular Drug Delivery; Necroptosis; Osteosarcoma Chemotherapy; Polypeptide Nanogel; Sarcoma-Targeting Peptide.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interest exists.

Figures

Scheme 1

Scheme 1

Schematic illustration for preparation of STP-NG/SHK, and RIP1- and RIP3-dependent cell necroptosis in primary tumor and lung metastasis.

Figure 1

Figure 1

Chemical structures and characterizations of untargeted and targeted SHK nanoformulations. Morphologies and _R_hs of (A) NG/SHK and (B) STP-NG/SHK. (C) Release profiles of SHK from NG/SHK and STP-NG/SHK in PBS without or with 10.0 mM GSH.

Figure 2

Figure 2

STP binds to osteosarcoma cells by CSV. (A) Cell-surface staining analysis for CSV in hFOB1.19 osteoblasts and 143B cells with CLSM. Scale bar = 25 μm. (B) Cells were stained with Anti-VIM conjugate 488 (green), STP-FITC (green), or Anti-VIM+ STP-FITC (green), and mean optical density (/pixel) was measured. Nuclei were stained with Hoechst 33342 (blue). Scale bar = 10 mm. (C) Immunologic assessment of STP binding to hFOB1.19 and 143B cells with FCM. Anti-VIM conjugate 488 was a positive control. Isotype controls were negative controls. Each set of data is represented as mean ± SEM (n = 3; &P < 0.001). (D) 143B cells were incubated with NG/SHK-FITC and STP-NG/SHK-FITC, and cellular uptake was measured by fluorecence imaging. Scale bar = 5 μm.

Figure 3

Figure 3

Cytotoxicity in vitro. (A) The cell viability of 143B cells was detected. (B) IC50s of SHK, NG/SHK, and STP-NG/SHK after 24 h. (C) Necroptosis in osteosarcoma 143B cells measured by FCM and Annexin V/7-AAD staining after treatments. Panel quadrants indicate populations of normal, early and late apoptotic, and necrotic cells. (D) RIP1, RIP3, PARP, caspase-3, and caspase-8 measurement via Western blot and (E) semiquantitative analyses. Each set of data is represented as mean ± SEM (n = 3; *P < 0.05, #P < 0.01, &P < 0.001).

Figure 4

Figure 4

Biodistribution. Biodistribution of SHK in 143B osteosarcoma-bearing mice at (A) 8 h and (B) 72 h after injection of SHK, NG/SHK, or STP-NG/SHK at a dose of 2.0 mg (kg BW)-1, intravenous injection. Each set of data is represented as mean ± SEM (n = 3; *P < 0.05, #P < 0.01, &P < 0.001).

Figure 5

Figure 5

In vivo antitumor efficacy of STP-NG/SHK against primary osteosarcoma. (A) Tumor growth curves. (B) Posterior limb tumors. (C) Average weight of tibial primary osteosarcoma tumors. (D) Tumor necrosis calculated with NIS-Elements imaging software. (E) H&E of primary tumor. Scale bar = 100 μm. (F) RIP1, RIP3, PARP, caspase-3, and caspase-8 expression in primary tumor tissues per group measured with Western blot and (G) semiquantitative analyses. Each set of data is represented as mean ± SEM (for A, C and D, n = 7, for G _n_=3; #P < 0.01, &P < 0.001).

Figure 6

Figure 6

In vivo antitumor efficiency of STP-NG/SHK against lung osteosarcoma metastasis. (A) Macroscopic appearance of lung metastatic osteosarcoma tumors per group. Scale bar = 1 cm. (B) Lung weight and (C) average microscopically (largest coronal sections) counted lung metastases. (D) H&E staining of metastases of osteosarcoma tumors. Arrows indicate lung metastases. Scale bar = 2 mm. (E) RIP1, RIP3, PARP, caspase-3, and caspase-8 expression in harvested lung metastases per group measured by Western blot and (F) semiquantitative analyses. Each set of data is represented as mean ± SEM (for B and C, n = 7; for E, n = 3; *P < 0.05, #P < 0.01, &P < 0.001).

Figure 7

Figure 7

Systemic toxicity of STP-NG/SHK against 143B osteosarcoma-bearing mice. (A) Weight changes. (B) Survival rates. (C) Mouse organs stained with H&E. Myocardia from mice in the SHK were damaged (★). No overt tissue damage was observed in STP-NG/SHK-treated mice. Scale bar = 100 μm (C). Each set of data is represented as mean ± SEM (for A, n = 7; *P < 0.05, #P < 0.01, &P < 0.001).

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