Novel strategies for personalized treatment of lung cancer using H19 gene regulatory sequences combined with targeted nucleic acid delivery
Final Report Abstract
Current treatment of advanced small-cell and non-small-cell lung cancer with chemotherapeutics is nonspecific, nonselective and toxic. Several tailored biological molecules are under clinical evaluation for targeted and personalized treatment regimes. We examined if BC-819, which is in clinical testing for bladder cancer, can further be used for lung cancer treatment. Furthermore, we analyzed the biological role of H19 RNA and its effectors that influence cancer development. We found, H19 is expressed in human lung tumors with elevated levels in more severe grades. Thus, H19 could serve as a qualified biomarker towards personalized lung cancer treatment. Aerosol application of BC-819 in an orthotopic lung cancer model was effective to prevent primary but not secondary lung tumors. Notably, it was encouraging, that repeated i.v. injections BC-819 decreased tumor loads in a metastatic lung tumor model. Likewise, various in vitro experiments suggested that H19 expression has a substantial stake in the invasiveness and the metastatic potential of lung cancer cells. The tumor suppressor p53 was identified as a negative regulator of the oncogene H19 and hypoxic stress as an inductor of H19 expression. Although, these results point to a pivotal role of H19 in cancer development, siRNA mediated knockdown failed to prevent tumor formation in vivo. In a second approach, we aimed to improve cell-specific mRNA activity in order to treat lung cancer more selectively. Incorporation of a microRNA binding site could be used to reduce mRNA expression in a cell specific manner. This approach may help to improve targeted mRNA expression upon in vivo application. In another attempt, we optimized nanoparticles to enhance selective targeting and to improve mRNA complexation. We found that i) grafting of polymers with PEG side chains increased mRNA binding and transfection efficiency remarkably, ii) insulin coating was useful to alter cell selectivity and iii) magnetized aerosols enable a localized treatment of defined lung areas with magnetized nanoparticles. Furthermore, we could show that dry powder aerosols can be used as efficient pulmonary gene delivery agents. Taken together, in this project we improved various parameters for gene delivery and addressed critical issues towards targeted and personalized lung cancer therapies.
Publications
- (2010). The oncofetal H19 RNA connection: hypoxia, p53 and cancer. Biochim Biophys Acta 1803(4):443-51
Matouk IJ, Mezan S, Mizrahi A, Ohana P, Abu-Lail R, Fellig Y, Degroot N, Galun E, Hochberg A
- (2011). Aerosolized BC-819 inhibits primary but not secondary lung cancer growth. PLoS One 6(6):e20760
Hasenpusch G, Pfeifer C, Aneja MK, Wagner K, Reinhardt D, Gilon M, Ohana P, Hochberg A, Rudolph C
- (2011). Dry powder aerosols of polyethylenimine (PEI)-based gene vectors mediate efficient gene delivery to the lung. J Control Release 154(1):69-76
Pfeifer C, Hasenpusch G, Uezguen S, Aneja MK, Reinhardt D, Kirch J, Schneider M, Claus S, Friess W, Rudolph C
- (2011). PEGylation improves nanoparticle formation and transfection efficiency of messenger RNA. Pharm Res. 28(9):2223-32
Uzgün S, Nica G, Pfeifer C, Bosinco M, Michaelis K, Lutz JF, Schneider M, Rosenecker J, Rudolph C
- (2012). Magnetized aerosols comprising superparamagnetic iron oxide nanoparticles improve targeted drug and gene delivery to the lung. Pharm Res. 29(5):1308-18
Hasenpusch G, Geiger J, Wagner K, Mykhaylyk O, Wiekhorst F, Trahms L, Heidsieck A, Gleich B, Bergemann C, Aneja MK, Rudolph C
(See online at https://doi.org/10.1007/s11095-012-0682-z) - (2013). The increasing complexity of the oncofetal h19 gene locus: functional dissection and therapeutic intervention. Int J Mol Sci. 14(2):4298-316
Matouk I, Raveh E, Ohana P, Lail RA, Gershtain E, Gilon M, De Groot N, Czerniak A, Hochberg A
