Abstract
The use of endogenous protoporphyrin IX after administration of 5-aminolaevulinic acid (ALA) has led to many applications in photodynamic therapy (PDT). We have previously reported that the conjugation of ALA dendrimers enhances porphyrin synthesis. The first aim of this work was to evaluate the ability of ALA dendrimers carrying 6 and 9 ALA residues (6m-ALA and 9m-ALA) to photosensitise cancer cells. For this aim, we employed LM3 mammary carcinoma cells. In these tumour cells, at low concentrations porphyrin synthesis from dendrimers was higher compared to ALA, whereas at high concentrations, porphyrin synthesis was similar from both compounds. Topical application of ALA dendrimers on the skin overlying a subcutaneous LM3 implanted tumour showed no diffusion of the molecules either to distant skin sites or to the adjacent tumour, suggesting a promising use of the ALA macromolecules in superficial cancer models. As a second objective, we proposed the use of ALA-dendrimers in vascular PDT for the treatment of atherosclerosis. Thus, we focused our studies on ALA-dendrimer’s selectivity towards macrophages in comparison with endothelial cells. For this aim we employed Raw 264.7 macrophages and HMEC-1 microvasculature cells. Porphyrin synthesis induced in macrophages by 6m-ALA and 9m-ALA (3 h, 0.025 mM) was 6 and 4.6 times higher respectively compared to the endothelial cell line, demonstrating the high affinity of ALA dendrimers for macrophages. On the other hand, ALA employed at low concentrations was slightly selective (1.7-fold) for macrophages. Inhibition studies suggested that ALA dendrimer uptake in macrophages is mainly mediated by caveloae-mediated endocytosis. Our main conclusion is that in addition to being promising molecules in PDT of superficial cancer, ALA dendrimers may also find applications in vascular PDT, since in vitro they showed selectivity to the macrophage component of the atheromatous plaque, as compared to the vascular endothelium.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- ALA:
-
5-Aminolevulinic acid
- MTT:
-
(3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazoliumbromide)
- PDT:
-
Photodynamic therapy
- PpIX:
-
Protoporphyrin IX
References
J. Kennedy, R. Pottier and G. Pross, Photodynamic Therapy with endogenous protoporphyrin IX: basic principles and present clinical experience, J. Photochem. Photobiol., B, 1990, 6, 143–148.
H. Fukuda, A. Casas, F. Chueke, S. Paredes and A. Batlle, Photodynamic action of endogenously syntesizedporphyrins from aminolevulinic acid, using a new model for assaying the effectiveness of tumoral cell killing, Int. J. Biochem., 1993, 25, 1395–1398.
J. Kopecek, P. Kopeckova, T. Minko and Z. Lu, HPMA copolymer anticancer drug conjugates: design, activity, and mechanism of action, Eur. J. Pharm. Biopharm., 2000, 50, 61–81.
R. Mehvar, Recent trends in the use of polysaccharides for improved delivery of therapeutic agents: pharmacokinetic and pharmacodynamic perspectives, Curr. Pharm. Biotechnol., 2003, 4, 283–302.
U. Boas and P. Heegaard, Dendrimers in drug research, Chem. Soc. Rev., 2004, 33, 43–63.
T. McCarthy, P. Karellas, S. Henderson, M. Giannis, D. O’Keefe, G. Heery, J. Paul, B. Matthews and G. Holan, Dendrimers as drugs: discovery and preclinical and clinicaldevelopment of dendrimer-based microbicides for HIV and STI prevention, Mol. Pharmacol., 2005, 2, 312–318.
S. Battah, C. Chee, H. Nakanishi, S. Gerscher, A. MacRobert and C. Edwards, Synthesis and biological studies of 5-aminolevulinic acid-containing dendrimers for photodynamic therapy, Bioconjugate Chem., 2001, 12, 980–988.
G. M. Di Venosa, A. G. Casas, S. Battah, P. Dobbin, H. Fukuda, A. J. Macrobert and A. Batlle, Investigation of a novel dendritic derivative of 5-aminolaevulinic acid for photodynamic therapy, Int. J. Biochem. Cell Biol., 2006, 38, 82–91.
S. Battah, S. Balaratnam, A. Casas, S. O’Neill, C. Edwards, A. Batlle, P. Dobbin and A. J. MacRobert, Macromolecular delivery of 5-aminolaevulinic acid for photodynamic therapy using dendrimer conjugates, Mol. Cancer Ther., 2007, 6, 876–885.
A. Casas, S. Battah, G. Di Venosa, P. Dobbin, L. Rodriguez, H. Fukuda, A. Batlle and A. J. Macrobert, Sustained and efficient porphyrin generation in vivo using dendrimer conjugates of 5-ALA for photodynamic therapy, J. Controlled Release, 2009, 135, 136–143.
S. G. Rockson, P. Kramer, M. Razavi, A. Szuba, S. Filardo, P. Fitzgerald, J. P. Cooke, S. Yousuf, A. R. DeVault, M. F. Renschler and D. C. Adelman, Photoangioplasty for human peripheral atherosclerosis: results of a phase I trial of photodynamic therapy with motexafin lutetium (Antrin), Circulation, 2000, 102, 2322–2324.
M. R. Hamblin and E. L. Newman, On the mechanism of the tumour-localising effect in photodynamic therapy, J. Photochem. Photobiol., B, 1994, 23, 3–8.
R. Straight, G. Vincent and E. Hammond, Porphyrin retention and photodynamic treatment of diet induced atherosclerotic lesions in pig, in Photodynamic Therapy of Tumors and Other Diseases, ed. G. Jori and C. Perria, LibreriaProgetto, Padova, Italy, 1986, pp. 350–352.
M. Eldar, Y. Yerushalmi and E. Kessler, Preferential uptake of a water soluble phthalocyanin by atherosclerotic plaques in rabbits, Atherosclerosis, 1990, 84, 135–139.
S. W. Young, K. W. Woodburn and M. Wright, Lutetium texaphyrin (PCI- 0123): a near-infrared, water-soluble photosensitizer, Photochem. Photobiol., 1996, 63, 892–897.
M. Miller, R. Kuntz and S. Friedrich, Frequency and consequences of intimal hyperplasia in specimens retrieved by directional atherectomy of native primary coronary artery stenoses and subsequent restenosis, Am. J. Cardiol., 1993, 71, 652–658.
S. G. Rockson, D. P. Lorenz, W. F. Cheong and K. W. Woodburn, Photoangioplasty: An emerging clinical cardiovascular role for photodynamic therapy, Circulation, 2000, 102, 591–596.
N. Kipshidze and H. Sahota, Photoangioplasty recount: clear punch or dimpled chad?, Circulation, 2001, 104, 55–56.
M. Pai, W. Jamal, A. Mosse, C. Bishop, S. Bown and J. R. McEwan, Inhibition of in-stent restenosis in rabbit iliac arteries with photodynamic therapy, Eur. J. Vasc. Endovasc. Surg., 2005, 30, 573–581.
M. Drakopoulou, K. Toutouzas, A. Michelongona, D. Tousoulis and C. Stefanadis, Vulnerable plaque and inflammation: potential clinical strategies, Curr. Pharm. Des., 2011, 17, 4190–4209.
M. P. Jenkins, G. A. Buonaccorsi, M. Raphael, I. Nyamekye, J. R. McEwan, S. G. Bown and C. C. R. Bishop, Clinical study of adjuvant photodynamic therapy to reduce restenosis following angioplasty, Br. J. Surg., 1999, 86, 1258–1263.
M. P. Jenkins, G. A. Buanoaccorsi, R. Mansfield, C. C. R. Bishop, S. G. Bown and J. R. McEwan, Reduction in the response to coronary and iliac artery injury with photodynamic therapy using 5-aminolevulinic acid, Cardiovasc. Res., 2000, 45, 478–485.
R. J. Mansfield, M. P. Jenkins, M. L. Pai, C. C. Bishop, S. G. Bown and J. R. McEwan, Long-term safety and efficacy of superficial femoral artery angioplasty with adjuvant photodynamic therapy to prevent restenosis, Br. J. Surg., 2002, 89, 1538–1539.
S. Werbajh, A. Urtreger, L. Puricelli, E. de Lustig, E. Bal de Kier Joffe and A. R. Kornblihtt, Downregulation of fibronectin transcription in highly metastatic adenocarcinoma cells, FEBS Lett., 1998, 440, 277–281.
E. W. Ades, F. J. Candal, R. A. Swerlick, V. G. George, S. Summers, D. C. Bosse and T. J. Lawley, HMEC-1: establishment of an immortalized human microvascular endothelial cell line, J. Invest. Dermatol., 1992, 99, 683–690.
F. Denizot and R. Lang, Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability, J. Immunol. Methods, 1986, 89, 271–277.
P. Workman, E. Aboagye, F. Balkwill, A. Balmain, G. Bruder, D. Chaplin, J. Double, J. Everitt, D. Farningham, M. Glennie, L. Kelland, V. Robinson, I. Stratford, G. Tozer, S. Watson, S. Wedge and S. Eccles, Committee of the National Cancer Research Institute, Guidelines for the welfare and use of animals in cancer research, Br. J. Cancer, 2010, 102, 1555–1577.
J. Van den Akker, V. Iani, W. Star, H. Sterenborg and J. Moan, Topical application of 5-aminolevulinic acid hexyl ester and 5-aminolevulinic acid to normal nude mouse skin: Differences in protoporphyrin IX fluorescence kinetics and the role of the stratum corneum, Photochem. Photobiol., 2000, 72, 681–689.
B. van Deurs, O. W. Petersen, S. Olsnes and K. Sandvig, Delivery of internalized ricin from endosomes to cisternal Golgi elements is a discontinuous, temperature-sensitive process, Exp. Cell Res., 1987, 171, 137–152.
W. A. Dunn, A. L. Hubbard and N. N. Aronson Jr., Low temperature selectively inhibits fusion between pinocytic vesicles and lysosomes during heterophagy of 125I-asialofetuin by the perfused rat liver, J. Biol. Chem., 1980, 255, 5971–5978.
M. Gunther, E. Wagner and M. Ogris, Specific targets in tumor tissue for the delivery of therapeutic genes, Curr. Med. Chem.: Anti-Cancer Agents, 2005, 5, 157–571.
A. François, S. Battah, A. J. MacRobert, L. Bezdetnaya, F. Guillemin and M. A. D’Hallewin, Fluorescence diagnosis of bladder cancer: a novel in vivo approach using 5-aminolevulinic acid (ALA) dendrimers, BJU Int., 2012, 110, 1155–1162.
C. Perotti, H. Fukuda, G. DiVenosa, A. J. MacRobert, A. Batlle and A. Casas, Porphyrin synthesis from ALA derivatives for photodynamic therapy. In vitro and in vivo studies, Br. J. Cancer, 2004, 90, 1660–1665.
G. Sahay, D. Y. Alakhova and A. V. Kabanov, Endocytosis of nanomedicines, J. Controlled Release, 2010, 145, 182–195.
K. M. Kitchens, A. B. Foraker, R. B. Kolhatkar, P. W. Swaan and H. Ghandehari, Endocytosis and interaction of poly (amidoamine) dendrimers with Caco-2 cells, Pharm. Res., 2007, 24, 2138–2145.
F. P. Seib, A. T. Jones and R. Duncan, Comparison of the endocytic properties of linear and branched PEIs, and cationic PAMAM dendrimers in B16f10 melanoma cells, J. Controlled Release, 2007, 117, 291–300.
T. G. Iversen, T. Skotland and K. Sandvig, Endocytosis and intracellular transport of nanoparticles: Present knowledge and need for future studies, Nano Today, 2011, 6, 176–185.
X. D. Zhu, Y. Zhuang, J. J. Ben, L. L. Qian, H. P. Huang, H. Bai, J. H. Sha, Z. G. He and Q. Chen, Caveolae-dependent endocytosis is required for class A macrophage scavenger receptor-mediated apoptosis in macrophages, J. Biol. Chem., 2011, 286, 8231–8239.
J. Mercer and A. Helenius, Virus entry by macropinocytosis, Nat. Cell Biol., 2009, 11, 510–520.
O. P. Perumal, R. Inapagolla, S. Kannan and R. M. Kannan, The effect of surface functionality on cellular trafficking of dendrimers, Biomaterials, 2008, 29, 3469–3476.
E. Fröhlich, The role of surface charge in cellular uptake and cytotoxicity of medical nanoparticles, Int. J. Nanomed., 2012, 7, 5577–5591.
M. Manunta, P. H. Tan, P. Sagoo, K. Kashefi, A.J George, Gene delivery by dendrimers operates via a cholesterol dependent pathway, Nucleic Acids Res., 2004, 32, 2730–2739.
M. Manunta, B. J. Nichols, P. H. Tan, P. Sagoo, J. Harper and A. J. George, Gene delivery by dendrimers operates via different pathways in different cells, but is enhanced by the presence of caveolin, J. Immunol. Methods, 2006, 314, 134–146.
K. Xiao, Y. Li, J. Luo, J. S. Lee, W. Xiao, A. M. Gonik, R. G. Agarwal and K. S. Lam, The effect of surface charge on in vivo biodistribution of PEG-oligocholic acid based micellarnanoparticles., Biomaterials, 2011, 32, 3435–3446.
S. Muro, M. Koval and V. Muzykantov, Endothelial endocytic pathways: gates for vascular drug delivery, Curr. Vasc. Pharmacol., 2004, 2, 281–299.
R. V. Stan, Endocytosis pathways in endothelium: how many?, Am. J. Physiol.: Lung Cell. Mol. Physiol., 2006, 290, 806–808.
M. G. Lei and D. C. Morrison, Differential expression of caveolin-1 in lipopolysaccharide-activated murine macrophages, Infect. Immun., 2000, 68, 5084–5089.
P. Gargalovic and L. Dory, Caveolin-1 and caveolin-2 expression in mouse macrophages. High density lipoprotein 3-stimulated secretion and a lack of significant subcellular co-localization, J. Biol. Chem., 2001, 276, 26164–26170.
I. A. Khalil, K. Kogure, H. Akita and H. Harashima, Uptake pathways and subsequent intracellular trafficking in nonviral gene delivery, Pharmacol. Rev., 2006, 58, 32–45.
T. N. Demidova and M. R. Hamblin, Macrophage-targeted photodynamic therapy, Int. J. Immunopathol. Pharmacol., 2004, 17, 117–126.
M. P. Jenkins, G. Buonaccorsi, A. MacRobert, C. C. R. Bishop, S. G. Bown and J. R. McEwarn, Intra-arterial photodynamic therapy using 5-ALA in a swine model, Eur. J. Vasc. Endovasc. Surg., 1998, 16, 284–291.
G. Di Venosa, H. Fukuda, A. Batlle, A. Macrobert and A. Casas, Photodynamic therapy: regulation of porphyrin synthesis and hydrolysis from ALA esters, J. Photochem. Photobiol., B, 2006, 83, 129–136.
C. Peng, Y. Li, H. Liang, J. Cheng, Q. Li, X. Sun, Z. Li, F. Wang, Y. Guo, Z. Tian, L. Yang, Y. Tian, Z. Zhang and W. Cao, Detection and photodynamic therapy of inflamed atherosclerotic plaques in the carotid artery of rabbits, J. Photochem. Photobiol., B., 2011, 102, 26–31.
O. C. Kwon, H. J. Yoon, K. H. Kim, H. T. Kim, Y. H. Yoon and J. K. Kim, Fluorescence kinetics of protoporphyrin-IX induced from 5-ALA compounds in rabbit postballoon injury model for ALA-photoangioplasty, Photochem. Photobiol., 2008, 84, 1209–1214.
Author information
Authors and Affiliations
Corresponding author
Additional information
Part of the data in this paper were presented during the 16th International Congress on Photobiology held in Cordoba, Argentina, in September (8th–12th), 2014.
Rights and permissions
About this article
Cite this article
Rodriguez, L., Vallecorsa, P., Battah, S. et al. Aminolevulinic acid dendrimers in photodynamic treatment of cancer and atheromatous disease. Photochem Photobiol Sci 14, 1617–1627 (2015). https://doi.org/10.1039/c5pp00126a
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1039/c5pp00126a