Bacteria-mediated tumor-targeted delivery of tumstatin (54-132) significantly suppresses tumor growth in mouse model by inhibiting angiogenesis and promoting apoptosis
1. The State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, Nanjing 210023, China 2. Changzhou High-Tech Research Institute of Nanjing University and Jiangsu Target Pharma Laboratories Inc., Changzhou 213164, China
Tumor growth is an angiogenesis-dependent process and accompanied by the formation of hypoxic areas. Tumstatin is a tumor-specific angiogenesis inhibitor that suppresses the proliferation and induces the apoptosis of tumorous vascular endothelial cells. VNP20009, an attenuated Salmonella typhimurium strain, preferentially accumulates in the hypoxic areas of solid tumors. In this study, a novel Salmonella-mediated targeted expression system of tumstatin (VNP-Tum5) was developed under the control of the hypoxia-induced J23100 promoter to obtain anti-tumor efficacy in mice. Treatment with VNP-Tum5 effectively suppressed tumor growth and prolonged survival in the mouse model of B16F10 melanoma. VNP-Tum5 exhibited a higher efficacy in inhibiting the proliferation and inducing the necrosis and apoptosis of B16F10 cells in vitro and in vivo compared with VNP (control). VNP-Tum5 significantly inhibited the proliferation and migration of mouse umbilical vascular endothelial cells to impede angiogenesis. VNP-Tum5 downregulated the expression of anti-vascular endothelial growth factor A, platelet endothelial cell adhesion molecule-1, phosphorylated phosphoinositide 3 kinase, and phosphorylated protein kinase B and upregulated the expression of cleaved-caspase 3 in tumor tissues. This study is the first to use tumstatin-transformed VNP20009 as a tumor-targeted system for treatment of melanoma by combining anti-tumor and anti-angiogenic effects.
J Jazowiecka-Rakus, S Szala. Antitumour activity of Salmonella typhimurium VNP20047 in B16(F10) murine melanoma model. Acta Biochim Pol 2004; 51( 3): 851– 856 https://doi.org/10.18388/abp.2004_3569
pmid: 15448746
3
KB Low, M Ittensohn, X Luo, LM Zheng, I King, JM Pawelek, D Bermudes. Construction of VNP20009: a novel, genetically stable antibiotic-sensitive strain of tumor-targeting Salmonella for parenteral administration in humans. Methods Mol Med 2004; 90 : 47– 60
pmid: 14657558
H Kimura, L Zhang, M Zhao, K Hayashi, H Tsuchiya, K Tomita, M Bouvet, J Wessels, RM Hoffman. Targeted therapy of spinal cord glioma with a genetically modified Salmonella typhimurium. Cell Prolif 2010; 43( 1): 41– 48 https://doi.org/10.1111/j.1365-2184.2009.00652.x
pmid: 19922490
6
X Luo Z Li S Lin T Le M Ittensohn D Bermudes JD Runyab SY Shen J Chen IC King LM Zheng. Antitumor effect of VNP20009, an attenuated Salmonella, in murine tumor models . Oncol Res 2001; 12(11−12): 501− 508
pmid: 11939414
7
JF Toso, VJ Gill, P Hwu, FM Marincola, NP Restifo, DJ Schwartzentruber, RM Sherry, SL Topalian, JC Yang, F Stock, LJ Freezer, KE Morton, C Seipp, L Haworth, S Mavroukakis, D White, S MacDonald, J Mao, M Sznol, SA Rosenberg. Phase I study of the intravenous administration of attenuated Salmonella typhimurium to patients with metastatic melanoma. J Clin Oncol 2002; 20( 1): 142– 152 https://doi.org/10.1200/JCO.2002.20.1.142
pmid: 11773163
8
NS Forbes, LL Munn, D Fukumura, RK Jain. Sparse initial entrapment of systemically injected Salmonella typhimurium leads to heterogeneous accumulation within tumors. Cancer Res 2003; 63( 17): 5188– 5193
pmid: 14500342
9
N Floquet, S Pasco, L Ramont, P Derreumaux, JY Laronze, JM Nuzillard, FX Maquart, AJ Alix, JC Monboisse. The antitumor properties of the α3(IV)-(185-203) peptide from the NC1 domain of type IV collagen (tumstatin) are conformation-dependent. J Biol Chem 2004; 279( 3): 2091– 2100 https://doi.org/10.1074/jbc.M307736200
pmid: 14583633
10
J Van der Velden, LM Harkness, DM Barker, GJ Barcham, CL Ugalde, E Koumoundouros, H Bao, LA Organ, A Tokanovic, JK Burgess, KJ Snibson. The effects of tumstatin on vascularity, airway inflammation and lung function in an experimental sheep model of chronic asthma. Sci Rep 2016; 6( 1): 26309 https://doi.org/10.1038/srep26309
pmid: 27199164
11
Y Maeshima, M Manfredi, C Reimer, KA Holthaus, H Hopfer, BR Chandamuri, S Kharbanda, R Kalluri. Identification of the anti-angiogenic site within vascular basement membrane-derived tumstatin. J Biol Chem 2001; 276( 18): 15240– 15248 https://doi.org/10.1074/jbc.M007764200
pmid: 11278365
12
T Kawaguchi, Y Yamashita, M Kanamori, R Endersby, KS Bankiewicz, SJ Baker, G Bergers, RO Pieper. The PTEN/Akt pathway dictates the direct αVβ3-dependent growth-inhibitory action of an active fragment of tumstatin in glioma cells in vitro and in vivo. Cancer Res 2006; 66( 23): 11331– 11340 https://doi.org/10.1158/0008-5472.CAN-06-1540
pmid: 17145879
13
SS Wahyu Effendi, SI Tan, WW Ting, IS Ng. Enhanced recombinant Sulfurihydrogenibium yellowstonense carbonic anhydrase activity and thermostability by chaperone GroELS for carbon dioxide biomineralization. Chemosphere 2021; 271 : 128461 https://doi.org/10.1016/j.chemosphere.2020.128461
pmid: 33131750
14
T Chen, X Zhao, Y Ren, Y Wang, X Tang, P Tian, H Wang, H Xin. Triptolide modulates tumour-colonisation and anti-tumour effect of attenuated Salmonella encoding DNase I. Appl Microbiol Biotechnol 2019; 103( 2): 929– 939 https://doi.org/10.1007/s00253-018-9481-8
pmid: 30448904
15
X Zhang, X Cheng, Y Lai, Y Zhou, W Cao, ZC Hua. Salmonella VNP20009-mediated RNA interference of ABCB5 moderated chemoresistance of melanoma stem cell and suppressed tumor growth more potently. Oncotarget 2016; 7( 12): 14940– 14950 https://doi.org/10.18632/oncotarget.7496
pmid: 26910836
16
XD Jiang, Y Qiao, P Dai, Q Chen, J Wu, DA Song, SQ Li. Enhancement of recombinant human endostatin on the radiosensitivity of human pulmonary adenocarcinoma A549 cells and its mechanism. J Biomed Biotechnol 2012; 2012 : 301931 https://doi.org/10.1155/2012/301931
pmid: 22778546
17
C Clairmont, KC Lee, J Pike, M Ittensohn, KB Low, J Pawelek, D Bermudes, SM Brecher, D Margitich, J Turnier, Z Li, X Luo, I King, LM Zheng. Biodistribution and genetic stability of the novel antitumor agent VNP20009, a genetically modified strain of Salmonella typhimurium. J Infect Dis 2000; 181( 6): 1996– 2002 https://doi.org/10.1086/315497
pmid: 10837181
18
J Sottile. Regulation of angiogenesis by extracellular matrix. Biochim Biophys Acta 2004; 1654( 1): 13– 22
pmid: 14984764
19
L Hlatky, P Hahnfeldt, J Folkman. Clinical application of antiangiogenic therapy: microvessel density, what it does and doesn’t tell us. J Natl Cancer Inst 2002; 94( 12): 883– 893 https://doi.org/10.1093/jnci/94.12.883
pmid: 12072542
20
W Li L Zhai Y Tang J Cai M Liu J Zhang. Antitumor properties of taxol in combination with cyclooxygenase-1 and cyclooxygenase-2 selective inhibitors on ovarian tumor growth in vivo. Oncol Res 2012; 20(2−3): 49− 59
pmid: 23193911
21
Y Maeshima, A Sudhakar, JC Lively, K Ueki, S Kharbanda, CR Kahn, N Sonenberg, RO Hynes, R Kalluri. Tumstatin, an endothelial cell-specific inhibitor of protein synthesis. Science 2002; 295( 5552): 140– 143 https://doi.org/10.1126/science.1065298
pmid: 11778052
22
Y Hamano, R Kalluri. Tumstatin, the NC1 domain of α3 chain of type IV collagen, is an endogenous inhibitor of pathological angiogenesis and suppresses tumor growth. Biochem Biophys Res Commun 2005; 333( 2): 292– 298 https://doi.org/10.1016/j.bbrc.2005.05.130
pmid: 15979458
23
Q Gu, T Zhang, J Luo, F Wang. Expression, purification, and bioactivity of human tumstatin from Escherichia coli. Protein Expr Purif 2006; 47( 2): 461– 466 https://doi.org/10.1016/j.pep.2006.01.011
pmid: 16503164
24
Y Hamano, M Zeisberg, H Sugimoto, JC Lively, Y Maeshima, C Yang, RO Hynes, Z Werb, A Sudhakar, R Kalluri. Physiological levels of tumstatin, a fragment of collagen IV α3 chain, are generated by MMP-9 proteolysis and suppress angiogenesis via αV β3 integrin. Cancer Cell 2003; 3( 6): 589– 601 https://doi.org/10.1016/S1535-6108(03)00133-8
pmid: 12842087
25
GM Zhang LH Sui T Jia YZ Zhao SB Fu XH Liu Y Yu. Inhibitory effect of recombinant anti-angiogenic peptide of tumstatin on growth and metastasis of human ovarian cancer transplanted in nude mice. Chin J Oncol (Zhonghua Zhong Liu Za Zhi) 2008; 30( 3): 170− 173 (in Chinese)
pmid: 18756929
26
A Sudhakar, CS Boosani. Inhibition of tumor angiogenesis by tumstatin: insights into signaling mechanisms and implications in cancer regression. Pharm Res 2008; 25( 12): 2731– 2739 https://doi.org/10.1007/s11095-008-9634-z
pmid: 18551250
27
R Weth, O Christ, S Stevanovic, M Zöller. Gene delivery by attenuated Salmonella typhimurium: comparing the efficacy of helper versus cytotoxic T cell priming in tumor vaccination. Cancer Gene Ther 2001; 8( 8): 599– 611 https://doi.org/10.1038/sj.cgt.7700352
pmid: 11571538
28
J Tjuvajev, R Blasberg, X Luo, LM Zheng, I King, D Bermudes. Salmonella-based tumor-targeted cancer therapy: tumor amplified protein expression therapy (TAPET) for diagnostic imaging. J Control Release 2001; 74( 1-3): 313– 315 https://doi.org/10.1016/s0168-3659(01)00340-6
pmid: 11489512
29
B Devraj, H Meenhard. Salmonella typhimurium as a novel RNA interference vector for cancer gene therapy. Cancer Biol Ther 2008; 7( 1): 151– 152 https://doi.org/10.416/cbt.7.15772
pmid: 1834715
30
S Mei, J Theys, W Landuyt, J Anne, P Lambin. Optimization of tumor-targeted gene delivery by engineered attenuated Salmonella typhimurium. Anticancer Res 2002; 22( 6A): 3261– 3266
pmid: 12530073
31
JT Bhushan, SF Neil. Motility is critical for effective distribution and accumulation of bacteria in tumor tissue. Integr Biol 2012; 4( 2): 165– 167 https://doi.org/10.1039/c2ib00091a
pmid: 22193245
32
KM Broadway, S Suh, B Behkam, BE Scharf. Optimizing the restored chemotactic behavior of anticancer agent Salmonella enterica serovar Typhimurium VNP20009. J Biotechnol 2017; 251 : 76– 83 https://doi.org/10.1016/j.jbiotec.2017.04.006
pmid: 28433721
33
DH Thamm, ID Kurzman, I King, Z Li, M Sznol, RR Dubielzig, DM Vail, EG MacEwen. Systemic administration of an attenuated, tumor-targeting Salmonella typhimurium to dogs with spontaneous neoplasia: phase I evaluation. Clin Cancer Res 2005; 11( 13): 4827– 4834 https://doi.org/10.1158/1078-0432.CCR-04-2510
pmid: 16000580