key: cord-0884831-fai1kymu authors: Abramczyk, H.; Brozek-Pluska, B.; Beton, Karolina title: Decoding COVID-19 mRNA Vaccine Immunometabolism in Central Nervous System: human brain normal glial and glioma cells by Raman imaging date: 2022-03-02 journal: bioRxiv DOI: 10.1101/2022.03.02.482639 sha: 027fc84b704910f3ef4992c9270dd225e56cd9dc doc_id: 884831 cord_uid: fai1kymu The paper presents the effect of COVID-19 mRNA (Pfizer/BioNT) vaccine on in vitro glial cells of the brain studied by means of Raman spectroscopy and imaging.. The results obtained for human brain normal and tumor glial cells of astrocytes, astrocytoma, glioblastoma incubated with the Covid-19 mRNA vaccine Pfizer/BioNT vaccine show alterations in the reduction-oxidation pathways associated with Cytochrome c. We found that the Pfizer/BioNT vaccine down regulate the concentration of cytochrome c in mitochondria upon incubation with normal and tumorous glial cells. Concentration of oxidized form of cytochrome c in brain cells has been shown to decrease upon incubation the mRNA vaccine. Lower concentration of oxidized cytochrome c results in lower effectiveness of oxidative phosphorylation (respiration), reduced apoptosis and lessened ATP production. Alteration of Amide I concentration, which may reflect the decrease of mRNA adenine nucleotide translocator. Moreover, mRNA vaccine leads to alterations in biochemical composition of lipids that suggest the increasing role of signaling. mRNA vaccine produce statistically significant changes in cell nucleus due to histone alterations. The results obtained for mitochondria, lipid droplets, cytoplasm may suggest that COVID-19 mRNA (Pfizer/BioNT) vaccine reprograms immune responses. The observed alterations in biochemical profiles upon incubation with COVID-19 mRNA in the specific organelles of the glial cells are similar to those we observe for brain cancer vs grade of aggressiveness. The COVID- 19 pandemics has witnessed an explosion in research in the field of 2 as a template for the production of viral proteins. These proteins are designed to trigger the production 48 of antibodies, which are then transferred to the host's immune system. response to mRNA vaccine in specific organelles without any labeling. 126 In this paper we will study implications for possible consequences of COVID-19 mRNA vaccine 127 (Pfizer/BioNT BNT162b2) on the central nervous system (CNS). We have already studied in detail the 128 biochemical alterations in specific organelles of brain cells vs cancer aggressiveness. [16, 17] Thus, we 129 can compare the effect of mRNA on normal and cancer cells with the effect of cancer aggressiveness. 130 As far as we know the mRNA Pfizer vaccine has not been tested for patients suffering of cancer. Therefore this contribution will help monitoring responses in host brain cells similar to a viral 132 infection, because the incubation with COVID-19 mRNA vaccine mimics COVID-19 infection, but 133 instead of the whole virus, only one key protein S for the immune response is synthesized, without 134 causing COVID-19 infection. 135 We will study human brain normal glial cells and glioma cells in vitro: normal human astrocytes causing COVID-19 infection. 141 We will monitor the effect of the mRNA vaccine on biodistribution of different chemical 142 components, particularly cytochrome c, in the specific organelles of a cell: nucleus, mitochondria, 143 lipid droplets, cytoplasm and membrane. 144 In the presented study we will identify dynamics and biochemical composition of the organelles 145 through characteristic Raman spectra upon injection of mRNA vaccine and incubation with the 146 vaccine in vitro cells. 147 We will show also that Raman spectroscopy and Raman imaging are competitive clinical 148 diagnostics tools for cancer diseases linked to mitochondrial dysfunction and are a prerequisite for 149 successful pharmacotherapy of cancer. 150 In this paper we explore alterations in reduction-oxidation pathways related to Cyt c in human 151 brain normal and tumor cells upon incubation in vitro with COVID-19 vaccine (Pfizer/ BioNT 152 BNT162b2). The studies were performed on normal human astrocytes (Clonetics NHA), human astrocytoma were recorded with a spatial resolution of 1 × 1 µm. The Raman microspectrometer was calibrated every 228 day prior to the measurements using a silica plate with a maximum peak at 520.7 cm -1 . The normalization was performed for low (500-1800 cm -1 ) and high (2600-3500 cm -1 ) frequency 238 spectral regions separately. For each organelle of a cell we have recorded hundreds of Raman spectra (because we have recorded and correlates with cancer aggressiveness [16] . We showed that the Raman Our results presented so far fully support these suggestions [17, 21] . triangle: cytochrome c-cancer -the immune system are strongly linked. 405 We found that there is very close relations between cytochrome c and 406 immune system through retinoic acid [22] . Retinoic acid (RA) is an essential 407 molecule in the innate immune system that does not stimulate its ATPase and 408 leads to lack of cytokine induction. Fig.6 shows effect of retinoic acid on in human lung cells [24] . (Figures 8 and 9 ) . RER is 466 studded with ribosomes that perform biological protein synthesis (mRNA 467 translation). Ribosomes are the sites of protein synthesis for mRNA vaccines. Ribosomes are too small to be seen by resolution of Raman imaging. with changes observed in mitochondria (Fig.5) . The alterations in lipid composition can also be monitored by the band at 492 2845 cm -1 that monitor concentration of triglycerides (TAG) [22, 25, 26] . Figure. Our results from Fig. 10 show that apoptosis is reduced upon mRNA vaccine We showed that new tools of Raman imaging we present in this paper raise exciting possibilities for new ways to understand links between pathways of cancer, immune responses, and recognize metabolites that regulates these pathways. We used Raman spectroscopy to monitor changes in the redox state of the mitochondrial cytochromes in human brain cells in vitro of normal astrocytes, astrocytoma, glioblastoma upon incubation with mRNA vaccine. We observed the effect of the mRNA vaccine on biodistribution of different chemical components, particularly cytochrome c, in the specific organelles of human brain glial cells: nucleus, mitochondria, lipid droplets, cytoplasm, rough endoplasmatic reticulum and membrane. We showed that mRNA vaccine (Pfizer) changes mitochondria by downregulation of cytochrome c resulting in lower effectiveness of respiration (oxidative phosphorylation) and lower ATP production. It can lead to lower immune system response. Decrease of Amide I concentration in mitochondrial membrane potential may suggest functional deterioration of the adenine nucleotide translocator. mRNA vaccine modifies significantly de novo lipids synthesis in lipid droplets. The results presented in paper suggest that upon incubation with mRNA the role of signaling function of lipid droplets increases. The observed alterations in biochemical profiles upon incubation with the Pfizer/BioNT in the specific organelles of the glial cells are similar to those we observe for brain cancer vs grade of aggressiveness. WHO Coronavirus (COVID-19) Dashboard | WHO Coronavirus Dashboard With Vaccination Data Available online Covid-19 Vaccines: In the Rush for Regulatory Approval, Do We Need More Data? SARS-CoV-2 MRNA Vaccines: Immunological Mechanism and Beyond Route of Vaccine Administration Alters Antigen Trafficking but Not Innate or Adaptive Immunity Preclinical and Clinical Demonstration of Immunogenicity by MRNA Vaccines against H10N8 and H7N9 Influenza Viruses Circulating Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Vaccine Antigen Detected in the Plasma of MRNA-1273 Vaccine Recipients The S1 Protein of SARS-CoV-2 Crosses the Blood-Brain Barrier in Mice The Neuroinvasive Potential of SARS-CoV2 May Play a Role in the Respiratory Failure of COVID-19 Patients The Involvement of the Central Nervous System in Patients with COVID-19 COVID-19 and the Brain: Acute Encephalitis as a Clinical Manifestation Prior Infection and Passive Transfer of Neutralizing Antibody Prevent Replication of Severe Acute Respiratory Syndrome Coronavirus in the Respiratory Tract of Mice Detection of Severe Acute Respiratory Syndrome Coronavirus in the Brain: Potential Role of the Chemokine Mig in Pathogenesis Infectability of Human BrainSphere Neurons Suggests Neurotropism of SARS-CoV-2 SARS-CoV-2 Infects Brain Astrocytes of COVID-19 Patients and Impairs Neuronal Viability The MRNA-LNP Platform's Lipid Nanoparticle Component Used in Preclinical Vaccine Studies Is Highly Inflammatory Revision of Commonly Accepted Warburg Mechanism of Cancer Development: Redox-Sensitive Mitochondrial Cytochromes in Breast and Brain Cancers by Raman Imaging Redox Imbalance and Biochemical Changes in Cancer by Probing Redox-Sensitive Mitochondrial Cytochromes in Label-Free Visible Resonance Raman Imaging. Cancers (Basel) Vaccine, MRNA) 4 Dosage And Administration | Pfizer Medical Information -Canada Available online Glucose Metabolism Inhibits Apoptosis in Neurons and Cancer Cells by Redox Inactivation of Cytochrome C A Walk in the Memory, from the First Functional Approach up to Its Regulatory Role of Mitochondrial Bioenergetic Flow in Health and Disease: Focus on the Adenine Nucleotide Translocator Cytochrome c as a Potentially Clinical Useful Marker of Mitochondrial and Cellular Damage Novel Strategies of Raman Imaging for Monitoring Intracellular Retinoid Metabolism in Cancer Cells Retinoic Acid: A Key Player in Immunity RIG-I Triggers a Signaling-Abortive Anti-SARS-CoV-2 Defense in Human Lung Cells A Look into the Use of Raman Spectroscopy for Brain and Breast Cancer Diagnostics: Linear and Non-Linear Optics in Cancer Research as a Gateway to Tumor Cell Identity Proteomic Analysis of Proteins Associated with Lipid Droplets of Basal and Lipolytically Stimulated 3T3-L1 Adipocytes SARS-CoV-2 Spike Protein Elicits Cell Signaling in Human Host Cells: Implications for Possible Consequences of COVID-19 Vaccines Resche-Rigon, M. The Mitochondrial Death/Life Regulator in Apoptosis and Necrosis Hallmarks of Cancer: The next Generation The Multiple Functions of Cytochrome c and Their Regulation in Life and Death Decisions of the Mammalian Cell: From Respiration to Funding: This work was supported by Statutory activity 2021: 501/3-34-1-1. The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.