Transplantation of bone marrow‑derived endothelial progenitor cells overexpressing Delta‑like‑4 enhances functional neovascularization in ischemic myocardium

Huang,Hong; Huang,Feng; Huang,Jian‑Ping

doi:10.3892/mmr.2013.1657

Transplantation of bone marrow‑derived endothelial progenitor cells overexpressing Delta‑like‑4 enhances functional neovascularization in ischemic myocardium

Authors:
- Hong Huang
- Feng Huang
- Jian‑Ping Huang
View Affiliations

Affiliations: Department of Cardiothoracic Surgery, The First People's Hospital of Nanning, Nanning, Guangxi 530022, P.R. China, Department of Emergency, The First People's Hospital of Nanning, Nanning, Guangxi 530022, P.R. China, Alibaba Business College, Hangzhou Normal University, Hangzhou, Zhejiang 310036, P.R. China
Published online on: August 28, 2013 https://doi.org/10.3892/mmr.2013.1657
Pages: 1556-1562

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Delta‑like‑4 (Dll‑4) prevents excess angiogenic sprouting and promotes the formation of a well‑differentiated vascular network. Therefore, transplantation of Dll‑4‑overexpressing endothelial progenitor cells (EPCs) was hypothesized to be superior to transplantation of EPCs in the treatment of ischemic heart disease. In the current study, EPCs harvested from C57BL/6 mouse bone marrow were infected in vitro with Dll‑4 (EPCDll‑4+) or Dll‑4 knockdown (EPCDll‑4‑) with recombinant lentiviral vectors and the control cells were non‑transfected or transduced with mock vectors (EPCnull). Eight‑week‑old C57BL/6 mice underwent ligation of the left anterior descending artery to establish a myocardial infarction (MI) model. The ligated animals were randomly divided into 5 groups, which, following one week, were intravenously injected with EPCs, EPCnull, EPCDll‑4+, EPCDll‑4‑ or medium. Two weeks later, echocardiographic assessment, western blotting, fluorescent microsphere and histological studies were performed. The results demonstrated that the number of mature vessels and blood flow in ischemic myocardium were increased in the EPCDll‑4+ group, but were markedly decreased in the EPCDll‑4‑ group compared with the control groups. The expression levels of Dll‑4, hairy/enhancer of split (Hes)‑related protein 1 (Hey‑1), phosphorylation of mammalian target of rapamycin (mTOR) and p70S6 kinase (p70S6K) were significantly increased in the EPCDll‑4+ group, while they were markedly decreased in the EPCDll‑4‑ group. Furthermore, for EPCDll‑4+‑treated animals, an enhanced cardiac function was observed as assessed by echocardiography. Thus, the transplantation of Dll‑4‑overexpressing EPCs stimulates neovascularization effectively, increases the blood flow to the ischemic zone and improves cardiac function. These effects may be due to the activation of Notch/Hey‑1/mTOR/p70S6K signaling pathways, which are initiated by Dll‑4.

View Figures

View References

1	Karamysheva AF: Mechanisms of angiogenesis. Biochemistry (Mosc). 73:751–762. 2008. View Article : Google Scholar
2	Lawall H, Bramlage P and Amann B: Treatment of peripheral arterial disease using stem and progenitor cell therapy. J Vasc Surg. 53:445–453. 2011. View Article : Google Scholar : PubMed/NCBI
3	Gimble JM, Bunnell BA and Guilak F: Human adipose-derived cells: an update on the transition to clinical translation. Regen Med. 7:225–235. 2012. View Article : Google Scholar : PubMed/NCBI
4	Kalka C, Masuda H, Takahashi T, Kalka-Moll WM, Silver M, Kearney M, Li T, Isner JM and Asahara T: Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization. Proc Natl Acad Sci USA. 97:3422–3427. 2000. View Article : Google Scholar : PubMed/NCBI
5	Asahara T, Murohara T, Sullivan A, Silver M, van der Zee R, Li T, Witzenbichler B, Schatteman G and Isner JM: Isolation of putative progenitor endothelial cells for angiogenesis. Science. 275:964–967. 1997. View Article : Google Scholar : PubMed/NCBI
6	Carmeliet P: Mechanisms of angiogenesis and arteriogenesis. Nat Med. 6:389–395. 2000. View Article : Google Scholar : PubMed/NCBI
7	Artavanis-Tsakonas S, Rand MD and Lake RJ: Notch signaling: cell fate control and signal integration in development. Science. 284:770–776. 1999. View Article : Google Scholar : PubMed/NCBI
8	Roca C and Adams RH: Regulation of vascular morphogenesis by Notch signaling. Genes Dev. 21:2511–2524. 2007. View Article : Google Scholar : PubMed/NCBI
9	Leslie JD, Ariza-McNaughton L, Bermange AL, McAdow R, Johnson SL and Lewis J: Endothelial signalling by the Notch ligand Delta-like 4 restricts angiogenesis. Development. 134:839–844. 2007. View Article : Google Scholar : PubMed/NCBI
10	Phng LK and Gerhardt H: Angiogenesis: a team effort coordinated by notch. Dev Cell. 16:196–208. 2009. View Article : Google Scholar : PubMed/NCBI
11	Scehnet JS, Jiang W, Kumar SR, et al: Inhibition of Dll4-mediated signaling induces proliferation of immature vessels and results in poor tissue perfusion. Blood. 109:4753–4760. 2007. View Article : Google Scholar : PubMed/NCBI
12	Noguera-Troise I, Daly C, Papadopoulos NJ, Coetzee S, Boland P, Gale NW, Lin HC, Yancopoulos GD and Thurston G: Blockade of Dll4 inhibits tumour growth by promoting non-productive angiogenesis. Nature. 444:1032–1037. 2006. View Article : Google Scholar : PubMed/NCBI
13	Ridgway J, Zhang G, Wu Y, et al: Inhibition of Dll4 signalling inhibits tumour growth by deregulating angiogenesis. Nature. 444:1083–1087. 2006. View Article : Google Scholar : PubMed/NCBI
14	Liu L, Wen T, Zheng XY, Yang DG, Zhao SP, Xu DY and Lü GH: Remnant-like particles accelerate endothelial progenitor cells senescence and induce cellular dysfunction via an oxidative mechanism. Atherosclerosis. 202:405–414. 2009. View Article : Google Scholar
15	Chen JJ and Zhou SH: Mesenchymal stem cells overexpressing MiR-126 enhance ischemic angiogenesis via the AKT/ERK-related pathway. Cardiol J. 18:675–681. 2011. View Article : Google Scholar : PubMed/NCBI
16	Urbich C, Aicher A, Heeschen C, Dernbach E, Hofmann WK, Zeiher AM and Dimmeler S: Soluble factors released by endothelial progenitor cells promote migration of endothelial cells and cardiac resident progenitor cells. J Mol Cell Cardiol. 39:733–742. 2005. View Article : Google Scholar : PubMed/NCBI
17	Huang F, Zhu X, Hu XQ, et al: Mesenchymal stem cells modified with miR-126 release angiogenic factors and activate Notch ligand Delta-like-4, enhancing ischemic angiogenesis and cell survival. Int J Mol Med. 31:484–492. 2013.PubMed/NCBI
18	Murohara T: Angiogenesis and vasculogenesis for therapeutic neovascularization. Nagoya J Med Sci. 66:1–7. 2003.PubMed/NCBI
19	Deuse T, Peter C, Fedak PW, et al: Hepatocyte growth factor or vascular endothelial growth factor gene transfer maximizes mesenchymal stem cell-based myocardial salvage after acute myocardial infarction. Circulation. 120(Suppl 11): S247–S254. 2009. View Article : Google Scholar
20	Suchting S, Freitas C, le Noble F, Benedito R, Bréant C, Duarte A and Eichmann A: The Notch ligand Delta-like 4 negatively regulates endothelial tip cell formation and vessel branching. Proc Natl Acad Sci USA. 104:3225–3230. 2007. View Article : Google Scholar : PubMed/NCBI
21	Lobov IB, Renard RA, Papadopoulos N, Gale NW, Thurston G, Yancopoulos GD and Wiegand SJ: Delta-like ligand 4 (Dll4) is induced by VEGF as a negative regulator of angiogenic sprouting. Proc Natl Acad Sci USA. 104:3219–3224. 2007. View Article : Google Scholar : PubMed/NCBI
22	Al Haj Zen A, Oikawa A, Bazan-Peregrino M, Meloni M, Emanueli C and Madeddu P: Inhibition of delta-like-4-mediated signaling impairs reparative angiogenesis after ischemia. Circ Res. 107:283–293. 2010.PubMed/NCBI
23	Miriuka SG, Rao V, Peterson M, Tumiati L, Delgado DH, Mohan R, Ramzy D, Stewart D, Ross HJ and Waddell TK: mTOR inhibition induces endothelial progenitor cell death. Am J Transplant. 6:2069–2079. 2006. View Article : Google Scholar : PubMed/NCBI
24	Maiese K, Chong ZZ, Shang YC and Hou J: FoxO proteins: cunning concepts and considerations for the cardiovascular system. Clin Sci (Lond). 116:191–203. 2009. View Article : Google Scholar : PubMed/NCBI
25	Lemaitre V, Dabo AJ and D’Armiento J: Cigarette smoke components induce matrix metalloproteinase-1 in aortic endothelial cells through inhibition of mTOR signaling. Toxicol Sci. 123:542–549. 2011. View Article : Google Scholar
26	Humar R, Kiefer FN, Berns H, Resink TJ and Battegay EJ: Hypoxia enhances vascular cell proliferation and angiogenesis in vitro via rapamycin (mTOR)-dependent signaling. FASEB J. 16:771–780. 2002. View Article : Google Scholar : PubMed/NCBI
27	Chan SM, Weng AP, Tibshirani R, Aster JC and Utz PJ: Notch signals positively regulate activity of the mTOR pathway in T-cell acute lymphoblastic leukemia. Blood. 110:278–286. 2007. View Article : Google Scholar : PubMed/NCBI
28	Shutter JR, Scully S, Fan W, Richards WG, Kitajewski J, Deblandre GA, Kintner CR and Stark KL: Dll4, a novel Notch ligand expressed in arterial endothelium. Genes Dev. 14:1313–1318. 2000.PubMed/NCBI
29	Hellström M, Phng LK, Hofmann JJ, et al: Dll4 signalling through Notch1 regulates formation of tip cells during angiogenesis. Nature. 445:776–780. 2007.PubMed/NCBI