Two distinct types of the inhibition of vasculogenesis by different species of charged particles

Journal of Angiogenesis Research

Charles Geard¹  Alan Bigelow²  Preety Sharma¹  Peter Grabham¹ 

[1] Center for Radiological Research, Columbia University, VC 11-205A/243, 630 West 168th street, New York, NY 10032, USA;Radiological Research Accelerator Facility, Center for Radiological Research, Nevis Laboratory, Columbia University, 136 S. Broadway, Irvington, NY 10533, USA

关键词: Motile tips;    3-D human vessel models;    Linear energy transfer;    Radiation;    Charged particle;    Vasculogenesis;   

Others  :  801504
DOI  :  10.1186/2045-824X-5-16

 received in 2013-07-08, accepted in 2013-09-02,  发布年份 2013

【 摘 要 】

Background

Charged particle radiation is known to be more biologically effective than photon radiation. One example of this is the inhibition of the formation of human blood vessels. This effect is an important factor influencing human health and is relevant to space travel as well as to cancer radiotherapy. We have previously shown that ion particles with a high energy deposition, or linear energy transfer (LET) are more than four times more effective at disrupting mature vessel tissue models than particles with a lower LET. For vasculogenesis however, the relative biological effectiveness between particles is the same. This unexpected result prompted us to investigate whether the inhibition of vasculogenesis was occurring by distinct mechanisms.

Methods

Using 3-Dimensional human vessel models, we developed assays that determine at what stage angiogenesis is inhibited. Vessel morphology, the presence of motile tip structures, and changes in the matrix architecture were assessed. To confirm that the mechanisms are distinct, stimulation of Protein Kinase C (PKC) with phorbol ester (PMA) was employed to selectively restore vessel formation in cultures where early motile tip activity was inhibited.

Results

Endothelial cells in 3-D culture exposed to low LET protons failed to make connections with other cells but eventually developed a central lumen. Conversely, cells exposed to high LET Fe charged particles extended cellular processes and made connections to other cells but did not develop a central lumen. The microtubule and actin cytoskeletons indicated that motility at the extending tips of endothelial cells is inhibited by low LET but not high LET particles. Actin-rich protrusive structures that contain bundled microtubules showed a 65% decrease when exposed to low LET particles but not high LET particles, with commensurate changes in the matrix architecture. Stimulation of PKC with PMA restored tip motility and capillary formation in low but not high LET particle treated cultures.

Conclusion

Low LET charged particles inhibit the early stages of vasculogenesis when tip cells have motile protrusive structures and are creating pioneer guidance tunnels through the matrix. High LET charged particles do not affect the early stages of vasculogenesis but they do affect the later stages when the endothelial cells migrate to form tubes.

【 授权许可】

   
2013 Grabham et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Wilson RR: Radiological use of fast protons. Radiology 1946, 47:487-491.
• [2]Cucinotta FA, Durante M: Cancer risk from exposure to galactic cosmic rays: implications for space exploration by human beings. Lancet Oncology 2006, 7:431-435.
• [3]Nelson GA: Fundamental space radiobiology. Gravit Space Biol Bull 2003, 16:29-36.
• [4]Townsend LW: Implications of the space radiation environment for human exploration in deep space. Radiat Prot Dosimetry 2005, 115:44-50.
• [5]Durante M, Kronenberg A: Ground-based research with heavy ions for space radiation protection. Adv Space Res 2005, 35:180-184.
• [6]Preston DL, Shimizu Y, Pierce DA, Suyama A, Mabuchi K: Studies of mortality of atomic bomb survivors. Report 13: solid cancer and noncancer disease mortality: 1950–1997. Rad Res 2003, 160:381-407.
• [7]Graebe A, Schuck EL, Lensing P, Putcha L, Derendorf H: Physiological, pharmacokinetic, and pharmacodynamic changes in space. J Clin Pharm 2004, 44:837-853.
• [8]Jurado JA, Bashir R, Mark W, Burket MW: Radiation-induced peripheral artery disease. Cath Card Interv 2008, 72:563-568.
• [9]Takahashi Y, Teshima T, Kawaguchi T, Hamada Y, Mori SA, Madachi A, Ikeda S, Mizuno H, Ogata T, Matsuura N: Heavy ion irradiation inhibits in vitro angiogenesis even at sublethal dose. Cancer Res 2003, 63:4253-4257.
• [10]Jang GH, Ha J-H, Huh T-L, Lee YM: Effect of proton beam on blood vessel formation in early developing zebrafish (Danio rerio) embryos. Arch Pharm Res 2008, 31:779-785.
• [11]Nelson GA, Rightnar S, Smith A, Jones T, Campbell-Beachler M, Mao XW, Favre C, Fike JR, Kubinova L, Anderson E: High-LET radiation-induced response of microvessels in the hippocampus. Rad Res 2011, 175:21-28.
• [12]Grabham P, Hu B, Sharma P, Geard C: Effects of ionizing radiation on three-dimensional human vessel models: differential effects according to radiation quality and cellular development. Rad Res 2011, 175:21-8.
• [13]Girdhani S, Lamont C, Hahnfeldt P, Abdollahi A, Hlatky L: Proton irradiation suppresses angiogenic genes and impairs cell invasion and tumor growth. Rad Res 2012, 178:33-45.
• [14]Davis GE, Camarillo CW: An alpha 2 beta 1 integrin-dependent pinocytic mechanism involving intracellular vacuole formation and coalescence regulates capillary lumen and tube formation in three-dimensional collagen matrix. Exp Cell Res 1996, 224:39-51.
• [15]Pate-Hett S, D’Amore PA: Signal transduction in vasculogenesis and developmental angiogenesis. Int J Dev Biol 2011, 55:353-363.
• [16]Bayless KJ, Johnson GA: Role of the cytoskeleton in formation and maintenance of angiogenic sprouts. J vasc res 2011, 48:369-385.
• [17]Grabham PW, Reznik B, Goldberg DJ: Microtubule and Rac 1-dependent F-actin in growth cones. J Cell Sci 2003, 116:3739-3748.
• [18]Oyanagi J, Ogawa T, Sato H, Higashi S, Miyazaki K: Epithelial-mesenchymal transition stimulates human cancer cells to extend microtubule-based invasive protrusions and suppresses cell growth in collagen gel. PLOS ONE 2012, 12:e53209.
• [19]Stratman AN, Saunders WB, Sacharidou A, Koh W, Fisher KE, Zawieja DC, Davis MJ, Davis GE: Endothelial cell lumen and vascular guidance tunnel formation requires MT1-MMP–dependent proteolysis in 3-dimensional collagen matrices. Blood 2009, 114:237-247.
• [20]Bayless KJ, Yeh AT, Lee PF: Nonlinear optical microscopy reveals invading endothelial cells anisotropically alter three-dimensional collagen matrices. Exp Cell Res 2009, 315:369-410.
• [21]Spalding AC, Zeitlin BD, Wilder-Romans K, Davis ME, Nor JE, Lawrence TS, Ben-Josef E: Enzastaurin, an inhibitor of PKC, enhances antiangiogenic effects and cytotoxicity of radiation against Endothelial Cells. Tranl Onc 2008, 1:195-201.
• [22]Willey CD, Xiao D, Tu T, Kim KW, Moretti L, Niermann KJ, Tawtawy MN, Quarles CC: Enzastaurin (LY317615), a protein kinase C beta selective inhibitor, enhances antiangiogenic effect of radiation. Int J Rad Oncol Biol Phys 2010, 77:1518-1526.
• [23]Girdhani S, Sachs R, Hlatky L: Biological effects of proton radiation: what we know and don’t know. Rad Res 2013, 179:257-272.
• [24]Durante M, Cucinotta FA: Heavy ion carcinogenesis and space exploration. Nat Rev Cancer 2008, 8:465-472.
• [25]Grabham P, Bigelow A, Geard C: DNA damage foci formation and decline in two-dimensional monolayers and in three-dimensional human vessel models: differential effects according to radiation quality. Int J Rad Biol 2012, 88:493-500.
• [26]Zeitlin C, Hassler DM, Cucinotta FA, Ehresmann B, Wimmer-Schweingruber RF, Brinza DE, Kang S, Weigle G, Bottcher S, Bohm E, Burmeister S, Guo J, Kohler J, Martin C, Posner A, Rafkin S, Reitz G: Measurements of energetic particle radiation in transit to mars on the mars science laboratory. Science 2013, 340:1080-1084.
• [27]NCRP: Information Needed to Make Radiation Protection Recommendations for Space Missions Beyond Low-Earth Orbit. Bethesda, MD: National Council on Radiation Protection and Measurements; 2006. Report No 153