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. 2024 Jan 31;25(3):1719.
doi: 10.3390/ijms25031719.

In Vitro MRS of Cells Treated with Trastuzumab at 1.5 Tesla

Wiesław Guz  1 , Rafal Podgórski  2 , Zuzanna Bober  3 , David Aebisher  3 , Adrian Truszkiewicz  3 , Marcin Olek  4 , Agnieszka Machorowska Pieniążek  4 , Aleksandra Kawczyk-Krupka  5 , Dorota Bartusik-Aebisher  2
Affiliations

In Vitro MRS of Cells Treated with Trastuzumab at 1.5 Tesla

Wiesław Guz et al. Int J Mol Sci. .

Abstract

The aim of the study was to investigate the effect of Trastuzumab on the MCF-7 and CRL-2314 breast cancer cell lines. Additionally, an attempt was made to optimize magnetic resonance spectroscopy (MRS) for cell culture studies, with particular emphasis on the impact of treatment with Trastuzumab. The research materials included MCF-7 and CRL-2314 breast cancer cell lines. The study examined the response of these cell lines to treatment with Trastuzumab. The clinical magnetic resonance imaging (MRI) system, OPTIMA MR360 manufactured by GEMS, with a magnetic field induction of 1.5 T, was used. Due to the nature of the tested objects, their size and shape, it was necessary to design and manufacture additional receiving coils. They were used to image the tested cell cultures and record the spectroscopic signal. The spectra obtained by MRS were confirmed by NMR using a 300 MHz NMR Fourier 300 with the TopSpin 3.1 system from Bruker. The designed receiving coils allowed for conducting experiments with the cell lines in a satisfactory manner. These tests would not be possible using factory-delivered coils due to their parameters and the size of the test objects, whose volume did not exceed 1 mL. MRS studies revealed an increase in the metabolite at 1.9 ppm, which indicates the induction of histone acetylation. Changes in histone acetylation play a very important role in both cell development and differentiation processes. The use of Trastuzumab therapy in breast cancer cells increases the levels of acetylated histones. MRS studies and spectra obtained from the 300 MHz NMR system are consistent with the specificity inherent in both systems.

Keywords: MRS; Trastuzumab; breast cancer.

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Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
The presentation of factors with influence on the development of Trastuzumab resistance.
Figure 2
Figure 2
MR spectra (a) and NMR spectra (b) of Trastuzumab.
Figure 3
Figure 3
MR Spectra (a) and NMR Spectra (b) of L-Histidine.
Figure 4
Figure 4
Representative 1H MRS of cells MCF-7 samples before treatment. The cells were properly cleaned by washing with PBS. There was no residual drug or culture medium left before preparing them for spectroscopy: 1—glutamine; 2—creatine; 3—total choline; 4—taurine; 5—glycine; 6—PCho; 7—GPC; 8—alanine.
Figure 5
Figure 5
(a) Representative 1H MRS of cells MCF-7 after treatment: 1—glutamine; 2—creatine; 3—total choline; 4—taurine; 5—glycine; 6—PCho; 7—GPC; 8—alanine; 9—Trastuzumab; 9a—pylosorbate 20; 9b—L-Histidine; 9c—D-Trehalose. (b) Spectrum of Trastuzumab.
Figure 6
Figure 6
MR Spectra for CRL-2314 (A) cells, CRL-2314 cells + 0.012 mg/mL Trastuzumab (B), CRL-2314 cells + 0.024 mg/mL Trastuzumab, (C) and CRL-2314 cells + 0.048 mg/mL Trastuzumab (D).
Figure 7
Figure 7
The ratio of choline to creatinine for breast cells CRL-2314 before and after treatment with the drug Trastuzumab dissolved in the culture medium.
Figure 8
Figure 8
MR Spectra for CRL-2314 (A) cells, CRL-2314 cells + 0.012 mg/mL Trastuzumab (B), CRL-2314 cells + 0.024 mg/mL Trastuzumab, (C) and CRL-2314 cells + 0.048 mg/mL Trastuzumab (D). The drug is dissolved in a liquid for injection.
Figure 9
Figure 9
The ratio of choline to creatinine for CRL-2314 cells before and after treatment with Trastuzumab dissolved in injection fluid.
Figure 10
Figure 10
Comparison of treatment results for CRL-2314 cells cultured in injection fluid and culture medium.
Figure 11
Figure 11
View of the bioreactor.
Figure 12
Figure 12
View of the receiving circuit during the test.
Figure 13
Figure 13
Result of the receiving coil analysis. RIGOL type: DSA815.
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