Tumor Biology

, Volume 37, Issue 8, pp 10643–10652 | Cite as

iRGD-targeted delivery of a pro-apoptotic peptide activated by cathepsin B inhibits tumor growth and metastasis in mice

  • Wang Qifan
  • Ning Fen
  • Xue Ying
  • Feng Xinwei
  • Du Jun
  • Zhang Ge
Original Article

Abstract

The use of cytolytic peptides with potential therapeutic properties is a promising approach to cancer therapy due to their convenient automated synthesis and their capacity for modifications. However, the use of cytolytic peptides is limited due to their nonspecific cytolytic activity. In this study, we designed a tumor-targeting proapoptotic system based on an amphipathic D-amino acid-modified apoptotic peptide, KLA, a variant of (KLAKLAK)2, which is fused with a linear tumor-penetrating homing peptide iRGD through specific cathepsin B (CTSB) cleavage sequences that are overexpressed in many types of tumor tissues. Our data show that the procytotoxic peptide D(KLAKLAKKLAKLA)K-GG-iRGD (m(KLA)-iRGD) is internalized into cultured tumor cells through a neuropilin-1 (NRP1)-activated pathway by iRGD delivery. Once inside the cells, the peptide triggers rapid apoptosis through both the mitochondrial-induced apoptotic pathway and the death receptor pathway in NRP1+/αvβ3/CTSB+ tumor cells. Furthermore, m(KLA)-iRGD spread extensively within the tumor tissue when it was injected into 4T1 tumor-bearing mice. The m(KLA)-iRGD peptide inhibited tumor growth to a certain degree, resulting in a significant reduction in tumor volume (P < 0.05) and the total inhibition of metastasis at the end of the treatment. These results suggest that m(KLA)-iRGD has the potential for development as a new antitumor drug.

Keywords

iRGD Cathepsin B Tumor growth Metastasis 

Notes

Acknowledgments

This work was supported by the National Natural Science Foundation of China (No. 81072670) and the Fundamental Research Funds for the Central Universities.

Compliance with ethical standards

Ethics statement

All of the animal procedures and experiments were approved by the Institutional Ethical Committee for Animal Research at Sun Yat-sen University. Twelve female BABL/c mice were used in the study.

Conflicts of interest

None

Supplementary material

13277_2016_4961_MOESM1_ESM.tif (349 kb)
Suppl. Fig 1 The chemical structure of m(KLA)-iRGD. The chemical structure of m(KLA)-iRGD (a) and control peptide D(KLA)-iRGD (b). m(KLA)-iRGD consists of a membrane-disrupting domain, a tumor-homing domain and a GG linker. The yellow domain represents the L-enantiomeric domain, and the blue domain represents the D-enantiomeric domain. The C-terminus of the yellow Lys is the cleavage site of CTSB. (TIF 348 kb)
13277_2016_4961_Fig7_ESM.gif (37 kb)

(GIF 37 kb)

13277_2016_4961_MOESM2_ESM.tif (5.4 mb)
Suppl. Fig 2 MS and HPLC analysis of m(KLA)-iRGD. a The molecular weight of m(KLA)- iRGD is 2570 Da. b The purity of m(KLA)-iRGD is approximately 95 %. (TIF 5573 kb)
13277_2016_4961_Fig8_ESM.gif (51 kb)

(GIF 50 kb)

13277_2016_4961_MOESM3_ESM.tif (504 kb)
Suppl. Fig 3 Expression levels of CTSB and NRP1 in different tumor cells. a The relative level of CTSB mRNA to GAPDH in different tumor cells measured by RT-PCR. CTSB mRNA shows higher expression in most tumor cell lines, except for SKBR3. b The expression levels of CTSB, NRP1, αv, β3 in MDA-MB-231, 4 T1, SKBR3 and B16 cell lines measured by Western blotting. (TIF 503 kb)
13277_2016_4961_Fig9_ESM.gif (60 kb)

(GIF 60 kb)

13277_2016_4961_MOESM4_ESM.tif (2 mb)
Suppl. Fig 4 Tumor tissues of the m(KLA)-iRGD group and the PBS group were removed and measured at the end of the experiments. a and b Tumor volumes of the m(KLA)-iRGD-treated mice were much smaller than those of the PBS control. (TIF 2084 kb)
13277_2016_4961_Fig10_ESM.gif (97 kb)

(GIF 97 kb)

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  • Wang Qifan
    • 1
  • Ning Fen
    • 2
  • Xue Ying
    • 1
  • Feng Xinwei
    • 1
  • Du Jun
    • 1
  • Zhang Ge
    • 1
  1. 1.Department of Microbial and Biochemical Pharmacy, School of Pharmaceutical SciencesSun Yat-sen UniversityGuangzhouChina
  2. 2.Guangzhou Institute of Pediatrics, Department of ObstetricsGuangzhou Women and Children’s Medical CenterGuangzhouChina

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