Biology subjects

Surmacki, J. M.

Publications and source records attributed to Surmacki, J. M..

5 recordsLinked to original sources

bioRxiv2024

The role of cytochrome c in mitochondrial metabolism of human oocytes

We investigated the biochemical composition of specific organelles in human oocyte cells at various maturation stages (GV, immature MI, MII with the first polar body, MII with giant polar body, and vacuoles) using Raman imaging. The structures analyzed included the nucleus, zona pellucida, perivitelline space, polar body, mitochondria, and cytoplasm. Raman imaging combined with chemometric classification via Cluster Analysis facilitated a comprehensive biochemical analysis of the proteomic, lipidomic, and DNA profiles of the human oocyte. Our findings indicate that cytochrome c is a crucial protein in oocyte mitochondria, essential for both cellular respiration (via oxidative phosphorylation) and apoptosis. Effective oocyte cell functioning requires cytochrome c in its redox-balanced forms: reduced and oxidized. The oxidized form of cytochrome c plays a pivotal role in the electron transport chain, enabling proper electron shuttling between complex III, cytochrome c, and complex IV. This process ensures controlled oxidative phosphorylation and ATP production, which are vital for most steps in oocyte maturation. BackgroundIn spite of progress in the identification of the fertile/infertile human oocytes in the recent decades, our understanding of molecular mechanisms occurring in the female reproductive cells did not make significant progress. ObjectiveThe present study aimed to develop our knowledge on mitochondrial metabolism in oocyte cells. ResultsBiochemical human oocyte analysis of proteomic-lipidomic-DNA profile by using Raman imaging combined with chemometric classification method of Cluster Analysis has been obtained. Our results show that cytochrome c is a key protein in oocyte mitochondria that is needed to maintain life (respiration via oxidative phosphorylation) and cell death (apoptosis). Materials and MethodsIn this experimental study, the nine fresh human native live oocytes from six donors collected in Salve Medica, Lodz, Poland have been investigated. The fingerprint and high-frequency region were evaluated by using Raman spectroscopy and imaging from 400 to 3500 cm-1. The principal component analysis method was used to visualize the difference in the Raman spectra of oocytes at different stage of maturation. The study was approved by the Bioethical Committee of the Medical University in Lodz, Poland (No RNN/83/23/KE). Written informed consent was obtained from all patients who voluntarily participated in the study. All procedures were conducted in accordance with the guidelines of the Declaration of Helsinki (2013). O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=119 SRC="FIGDIR/small/616010v1_ufig1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@db4837org.highwire.dtl.DTLVardef@623424org.highwire.dtl.DTLVardef@14d6748org.highwire.dtl.DTLVardef@b7b1de_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗

bioRxiv2024

A novel HER2 protein identification methodology in breast cancer cells using Raman spectroscopy and Raman imaging: an analytical validation study

BackgroundConventional assays such as immunohistochemistry (IHC) and in situ hybridization (ISH) used in clinical procedures for quantification of the human epidermal growth factor receptor-2 (HER2) status in breast cancer have many limitations. Our results suggest that a new Raman method may improve specificity that will result in better patients selection for HER2 targeted therapy. In the current study, we have used HER2 expression in a broad range of breast cancer phenotypes to explore the potential utility of a novel immunodetection technique, using Raman spectroscopy and Raman imaging combined with artificial intelligence models. MethodsThe expression of HER2 protein in different cancer subtypes was evaluated using Raman methodology to test correlations with currently used quantitative protein analysis for a broad range of cancer subtypes based on the expression levels of estrogen and progesterone receptors (ER and PR), HER2, cytokeratins and claudins. In the current study, five breast cancer cell lines: MCF-10A, MCF-7, MDA-MB-231, HTB-30 (SK-BR-3) and AU-565 were tested. ResultsThe correlations between Raman method and conventional HER2 testing methodologies (IHC and ISH) have been tested. Raman measurements showed a strong linear correlation (p= 0.05, R2=0,9816) with IHC analysis in the studied breast cell lines: MCF-10A, MCF-7, MDA-MB- 231, HTB-30 (SK-BR-3) and AU-565 representing normal, non-tumorigenic epithelial cells, triple-positive breast carcinoma and triple-negative breast cancer cell lines. ConclusionsAnalytic testing of Raman spectroscopy and Raman imaging demonstrated that this method may offer advantages over currently used diagnostic methodologies. The broad range of HER2 expression on the surface of human breast normal and cancer cells from triple negative to triple positive cell lines were studied. Our data demonstrate that Raman based methods for HER2 quantitation of HER2 may offer significant progress in patient selection for HER2 targeted therapy over conventional HER2 identification. Further studies of HER2 determination by Raman methods with in vivo cells and ex vivo tissue are warranted.

cancer biology↗

bioRxiv2022

The effect of COVID-19 mRNA vaccine on respiratory system: human lung carcinoma cells by means of Raman spectroscopy and imaging.

The effect of COVID-19 mRNA vaccine on human lung carcinoma epithelial cells (A549) in vitro as a convenient preclinical model has been studied by means of Raman spectroscopy and imaging. The paper focuses on Raman imaging as a tool to explore apoptosis and oxidative phosphorylation in mitochondrial dysfunctions. The Raman results demonstrate alterations in the oxidation-reduction pathways associated with cytochrome c. We found that the COVID-19 mRNA vaccine downregulates the concentration of cytochrome c upon incubation with tumorous lung cells. Concentration of oxidized form of cytochrome c in mitochondria of lung cells decreases upon incubation the COVID-19 mRNA vaccine. Lower concentration of oxidized cytochrome c in mitochondria illustrates lower effectiveness of oxidative phosphorylation (respiration), reduced apoptosis and lessened ATP production. Moreover, mRNA vaccine increases significantly de novo lipids synthesis in lipid droplets up to 96 hours and alterations in biochemical composition. It seems that lipid composition of cells returns to the normal level for longer incubation time (14 days). In cell nucleus the mRNA vaccine does not produce statistically significant changes. The observed alterations in biochemical profiles upon incubation with mRNA vaccine in the specific organelles of the tumorous lung cells are similar to those we observe for other types of cancers, particularly brain glial cells.

cell biology↗

bioRxiv2020

Redox state changes of mitochondrial cytochromes in brain and breast cancers by Raman spectroscopy and imaging

This paper presents a non-invasive approach to study redox status of cytochromes in vitro human brain cells of normal astrocytes (NHA), astrocytoma (CRL-1718), glioblastoma (U87- MG) and medulloblastoma (Daoy), and human breast cells of normal cells (MCF10A), slightly malignant cells (MCF7) and highly aggressive cells (MDA-MB-231), in vivo animal models, and ex vivo brain and breast tissues surgically resected human specimens by means of Raman microspectroscopy at 355 nm, 532 nm, 785 nm and endospectroscopic Raman probe at 785 nm. Here we show that the amount of reduced cytochrome becomes abnormally high in human brain tumors and breast cancers. In contrast, the amount of reduced cytochrome c is lower in cancer cells when compared to the normal one at in vitro conditions when the effect of microenvironment is eliminated. Mitochondrial dysfunction and alterations in the chemical composition of the nucleus, mitochondria, lipid droplets, cytoplasm in single cells have been detected by Raman imaging. Incubation in vitro with retinoic acid increases the amount of reduced cytochrome c.

cancer biology↗

bioRxiv2020

Novel strategies of Raman imaging for monitoring intracellular retinoid metabolism in cancer cells

We developed a label-free Raman method for whole-cell biochemical imaging to detect molecular processes that occur in normal and cancer brain cells due to retinol transport in human cancers at the level of isolated organelles. Our approach allows to create biochemical maps of retinoids localization in lipid droplets, mitochondria and nuclei in single cells. The maps were capable of discriminating triglycerides (TAG) from retinoids (RE) in lipid droplets (LD), and mitochondria providing an excellent tool to monitor intracellular retinoid metabolism. We detected spectral changes that arose in proteins and lipids due to retinoid metabolism in human cell lines of normal astrocytes and high-grade cancer cells of glioblastoma as well as in human medulloblastoma and glioblastoma tissue. Raman imaging is an effective tool for monitoring retinoids and retinol binding proteins involved in carcinogenesis by detecting unique spectral signatures of vibrations. We found two functionally distinct lipid droplets: TAG-LD, for energy storage, and RE-LD, for regulating mechanisms of signal transduction. Raman polarization measurements revealed the occurrence of conformational changes affecting discrete regions of proteins associated with retinol binding. Aberrant expression of retinoids and retinol binding proteins in human tumours were localized in lipid droplets, and mitochondria.

cancer biology↗