Study of the intracellular nanoparticle-based radiosensitization mechanisms in F98 glioma cells treated with charged particle therapy through synchrotron-based infrared microspectroscopy

Abstract

The use of nanoparticles (NP) as dose enhancers in radiotherapy (RT) is a growing research field. Recently, the use of NP has been extended to charged particle therapy in order to improve the performance in radioresistant tumors. However, the biological mechanisms underlying the synergistic effects involved in NP-RT approaches are not clearly understood. Here, we used the capabilities of synchrotron-based Fourier Transform Infrared Microspectroscopy (SR-FTIRM) as a bio-analytical tool to elucidate the NPinduced cellular damage at the molecular level and at a single-cell scale. F98 glioma cells doped with AuNP and GdNP were irradiated using several types of medical ion beams ( proton, helium, carbon and oxygen). Differences in cell composition were analyzed in the nucleic acids, protein and lipid spectral regions using multivariate methods (Principal Component Analysis, PCA). Several NP-induced cellular modifications were detected, such as conformational changes in secondary protein structures, intensity variations in the lipid CH x stretching bands, as well as complex DNA rearrangements following charged particle therapy irradiations. These spectral features seem to be correlated with the already shown enhancement both in the DNA damage response and in the reactive oxygen species (ROS) production by the NP, which causes cell damage in the form of protein, lipid, and/or DNA oxidations. Vibrational features were NP-dependent due to the NP heterogeneous radiosensitization capability. Our results provided new insights into the molecular changes in response to NP-based RT treatments using ion beams, and highlighted the relevance of SR-FTIRM as a useful and precise technique for assessing cell response to innovative radiotherapy approaches. a MIRAS beamline BL01, ALBA-CELLS Synchrotron,

Graphene Oxide-Induced Protein Conformational Change in Nasopharyngeal Carcinoma Cells: A Joint Research on Cytotoxicity and Photon Therapy

“…Holman et al [46] normalized IR spectra (of normal human fetal lung fibroblast) to amide I peak intensity, and discovered that the amide II band increased, along with peak wavelength shifting downward, for dying cell as opposed to normal living cell. They ascribed the peak shift to lower wavelength to the protein structural change from α-helix to β-sheet [46,48,49]. Hence, both amide I and amide II peak shift and their intensity variation in GO-treated sample confirms the change in the secondary structure of proteins, as compared with pristine NPC-BM1 cells.…”

“…The cytotoxicity of control and nanometric GO-treated NPC-BM1 cells was studied in a dose-dependent manner, and the data were reported as viable cell percentages. The GO concentration-dependent cytotoxic effects (0.1-10 µg mL −1 ) of NPC-BM1 cells at different time periods (24,48, and 72 h) are presented in Figure 4. No noteworthy reduction in their metabolic activity at first 24 h was observed after exposure to the lower GO concentration (0.1-0.5 µg mL −1 ) compared with the control.…”

“…Figure 6a shows that there were significant peak intensity increases in the NPC-BM1 cells after the GO treatment, particularly in the amide I and amide II functional groups (at 1644 and 1536 cm −1 , respectively). These amide groups are related to protein components [46][47][48]. The amide I groups consists of C=O stretching in protein molecules, whereas amide II represents the δN-H bending and vC-N stretching vibrations [43].…”

Nanoparticles for Stem Cell Therapy Bioengineering in Glioma

“…2019; Shi et al, 2019;Sousa et al, 2019;Ye et al, 2019;Zhang et al, 2019;Zhao et al, 2019;ZhuGe et al, 2019;Alves et al, 2020;Caban-Toktas et al, 2020;Chung et al, 2020;Ferreira et al, 2020;Kazmi et al, 2020;Liang et al, 2020;Martinez-Rovira et al, 2020;Meng et al, 2020;Roberts et al, 2020;Ullah et al, 2020;Wang H. et al, 2020;Wang X. et al, 2020;Xu et al, 2020;Zhu et al, 2020; see Table 3 for examples of ligand targeting in glioma research). Another active targeting method to increase functional specificity of NPs that are used as gene delivery systems is the transcriptional targeting, which can occur at a transcriptional or posttranscriptional level (Golombek et al, 2018).…”

“…Here, the delivered gene includes a tumor-specific promoter (highly functional only in cancer cells), which will secure a well-localized expression of the transgene, limited to occur only inside the cancer cells of interest. Posttranscriptional regulations of the product encoded by the exogenously delivered gene are achieved by controlling RNA splicing, RNA stability, and initiation of the RNA translation once it is present in the cancer cell (Maier-Hauff et al, 2007;Kim et al, 2010;Wegscheid et al, 2014;Cheng Y. et al, 2016;Shen et al, 2017;Yu et al, 2017;Hua et al, 2018;Daniel et al, 2019;Denora et al, 2019;Dufort et al, 2019;Kunoh et al, 2019;Luque-Michel et al, 2019;Ruan et al, 2019;Rego et al, 2019Rego et al, , 2020Shi et al, 2019;Sousa et al, 2019;Ye et al, 2019;Zhang et al, 2019;Zhao et al, 2019;ZhuGe et al, 2019;Alves et al, 2020;Caban-Toktas et al, 2020;Chung et al, 2020;Ferreira et al, 2020;Kazmi et al, 2020;Liang et al, 2020;Martinez-Rovira et al, 2020;Meng et al, 2020;Roberts et al, 2020;Ullah et al, 2020;Wang H. et al, 2020;Wang X. et al, 2020;Xu et al, 2020;Zhu et al, 2020).…”