scholarly journals Cell‐Free Mitochondrial DNA as a Potential Biomarker for Astronauts' Health

2021 ◽  
Author(s):  
Malik Bisserier ◽  
Santhanam Shanmughapriya ◽  
Amit Kumar Rai ◽  
Carolina Gonzalez ◽  
Agnieszka Brojakowska ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Mononuclear Cells ◽  
Physiological Stress ◽  
Risk Mitigation ◽  
Space Station ◽  
Mitigation Strategies ◽  
Diagnostic Tools ◽  
Peripheral Blood Mononuclear ◽  
Markers Of Inflammation ◽  
Potential Biomarker

Background Space travel–associated stressors such as microgravity or radiation exposure have been reported in astronauts after short‐ and long‐duration missions aboard the International Space Station. Despite risk mitigation strategies, adverse health effects remain a concern. Thus, there is a need to develop new diagnostic tools to facilitate early detection of physiological stress. Methods and Results We measured the levels of circulating cell‐free mitochondrial DNA in blood plasma of 14 astronauts 10 days before launch, the day of landing, and 3 days after return. Our results revealed a significant increase of cell‐free mitochondrial DNA in the plasma on the day of landing and 3 days after return with vast ~2 to 355‐fold interastronaut variability. In addition, gene expression analysis of peripheral blood mononuclear cells revealed a significant increase in markers of inflammation, oxidative stress, and DNA damage. Conclusions Our study suggests that cell‐free mitochondrial DNA abundance might be a biomarker of stress or immune response related to microgravity, radiation, and other environmental factors during space flight.

scholarly journals LPS-Induced Endotoxemia Evokes Epigenetic Alterations in Mitochondrial DNA That Impacts Inflammatory Response

Cells ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 2282
Author(s):  
Björn Koos ◽  
Eva Lotta Moderegger ◽  
Katharina Rump ◽  
Hartmuth Nowak ◽  
Katrin Willemsen ◽  
...  
Keyword(s):  
Immune Response ◽  
Mitochondrial Dna ◽  
Dna Methyltransferase ◽  
Protein Import ◽  
Mononuclear Cells ◽  
Mitochondrial Protein ◽  
Mitochondrial Fraction ◽  
Protein Abundance ◽  
Peripheral Blood Mononuclear ◽  
Cellular Expression

Mitochondrial DNA (mtDNA) plays a vital role as a damage-associated molecular pattern in sepsis being able to shape the immune response. Since pathogen recognition receptors of innate immune cells are activated by demethylated DNA only, we set out to investigate the amount of DNA methyltransferase 1 (DNMT1) in mitochondria and the extent of mtDNA methylation in a human endotoxin model. Peripheral blood mononuclear cells of 20 healthy individuals were isolated from whole blood and stimulated with lipopolysaccharide (LPS) for 48 h. Subsequently, DNMT1 protein abundance was assessed in whole cells and a mitochondrial fraction. At the same time, methylation levels of mtDNA were quantified, and cytokine expression in the supernatant was measured. Despite increased cellular expression of DNMT1 after LPS stimulation, the degree of mtDNA methylation slightly decreased. Strikingly the mitochondrial protein abundance of DNMT1 was reduced by 50% in line with the lower degree of mtDNA methylation. Although only modest alterations were seen in the degree of mtDNA methylation, these strongly correlated with IL-6 and IL-10 expression. Our data may hint at a protein import problem for DNMT1 into the mitochondria under LPS stimulation and suggest a role of demethylated mtDNA in the regulation of the inflammatory immune response.

Frataxin deficiency in Friedreich’s ataxia is associated with reduced levels of HAX-1, a regulator of cardiomyocyte death and survival

2020 ◽  
Vol 29 (3) ◽  
pp. 471-482
Author(s):  
Francesca Tiano ◽  
Francesca Amati ◽  
Fabio Cherubini ◽  
Elena Morini ◽  
Chiara Vancheri ◽  
...  
Keyword(s):  
Pancreatic Beta Cells ◽  
Mononuclear Cells ◽  
Premature Death ◽  
Friedreich’S Ataxia ◽  
Friedreich's Ataxia ◽  
Peripheral Blood Mononuclear ◽  
Protein Levels ◽  
Potential Biomarker ◽  
Mrna And Protein Expression ◽  
Defence Proteins

Abstract Frataxin deficiency, responsible for Friedreich’s ataxia (FRDA), is crucial for cell survival since it critically affects viability of neurons, pancreatic beta cells and cardiomyocytes. In FRDA, the heart is frequently affected with typical manifestation of hypertrophic cardiomyopathy, which can progress to heart failure and cause premature death. A microarray analysis performed on FRDA patient’s lymphoblastoid cells stably reconstituted with frataxin, indicated HS-1-associated protein X-1 (HAX-1) as the most significantly upregulated transcript (FC = +2, P < 0.0006). quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) and western blot analysis performed on (I) HEK293 stably transfected with empty vector compared to wild-type frataxin and (II) lymphoblasts from FRDA patients show that low frataxin mRNA and protein expression correspond to reduced levels of HAX-1. Frataxin overexpression and silencing were also performed in the AC16 human cardiomyocyte cell line. HAX-1 protein levels are indeed regulated through frataxin modulation. Moreover, correlation between frataxin and HAX-1 was further evaluated in peripheral blood mononuclear cells (PBMCs) from FRDA patients and from non-related healthy controls. A regression model for frataxin which included HAX-1, group membership and group* HAX-1 interaction revealed that frataxin and HAX-1 are associated both at mRNA and protein levels. Additionally, a linked expression of FXN, HAX-1 and antioxidant defence proteins MnSOD and Nrf2 was observed both in PBMCs and AC16 cardiomyocytes. Our results suggest that HAX-1 could be considered as a potential biomarker of cardiac disease in FRDA and the evaluation of its expression might provide insights into its pathogenesis as well as improving risk stratification strategies.

IMPACT: Immunotherapy in patients with metastatic cancers and CDK12 mutations.

2019 ◽  
Vol 37 (15_suppl) ◽  
pp. TPS5091-TPS5091 ◽  
Author(s):  
Melissa Andrea Reimers ◽  
Wassim Abida ◽  
Jonathan Chou ◽  
Daniel J. George ◽  
Elisabeth I. Heath ◽  
...  
Keyword(s):  
T Cell ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Immune Cell ◽  
Castration Resistant Prostate Cancer ◽  
Open Label ◽  
Histologic Diagnosis ◽  
Peripheral Blood Mononuclear ◽  
Diversity Assessment ◽  
Potential Biomarker

TPS5091 Background: Tumors with biallelic CDK12 loss have been identified as a distinct subtype in metastatic castration resistant prostate cancer (mCRPC) and other cancer types. The CDK12 biallelic loss mCRPC genomic signature, distinct from homologous recombination deficient (HRD) and ETS fusion signatures, is characterized by excessive tandem duplications, genomic instability, gene fusion-caused putative neoantigens, and increased tumor T cell infiltration. Early clinical experience with anti-PD-1 immunotherapy in CDK12 loss mCRPC patients (pts) is notable for deep and sustained PSA as well as radiographic responses. We hypothesize that CDK12 biallelic loss is a potential biomarker of immune checkpoint immunotherapy (ICI) efficacy in mCRPC and other cancers. Methods: IMPACT (NCT03570619) is a multi-center, open label, phase 2 study of pts with metastatic cancers that harbor CDK12 biallelic loss. mCRPC pts will be enrolled in cohort A (n = 25) in a Mini-Max Simon Two-Stage design, and all other pts in single-stage cohort B (n = 15). All pts will receive induction therapy with nivolumab 3 mg/kg IV and ipilimumab 1 mg/kg IV q3 weeks for up to 4 cycles, followed by maintenance nivolumab at 480 mg IV q4 weeks (up to 52 weeks in total). Eligible pts must have identified biallelic CDK12 loss on any CLIA/CAP approved next generation sequencing assay and a histologic diagnosis of metastatic prostate adenocarcinoma or other metastatic carcinoma. No prior ICI is allowed. The primary endpoint is the overall response rate (ORR) in cohort A per PCWG3 criteria. An ORR of 30% is targeted in cohort A. Secondary endpoints include safety, secondary efficacy measures, quality of life, and survival measures. Exploratory objectives include tumor whole exome analysis and changes in immune profiles with therapy. Comprehensive and serial monitoring of peripheral blood immune cell populations will be performed via T cell clonal diversity assessment and multi-parametric flow cytometry. Changes in myeloid and lymphoid populations will be assessed from whole blood. Polarization and effector function of T cells and activation of antigen presenting cells will be further characterized from isolated peripheral blood mononuclear cells. Study accrual is ongoing. Clinical trial information: NCT03570619.

scholarly journals Development of a quantitative PCR assay for measurement of trichechid herpesvirus 1 load in the Florida manatee (Trichechus manatus latirostris)

2017 ◽  
Vol 29 (4) ◽  
pp. 476-482 ◽  
Author(s):  
Jason A. Ferrante ◽  
Galaxia Cortés-Hinojosa ◽  
Linda L. Archer ◽  
James F. X. Wellehan
Keyword(s):  
Mononuclear Cells ◽  
Qpcr Assay ◽  
Pcr Assay ◽  
Florida Manatee ◽  
Peripheral Blood Mononuclear ◽  
Trichechus Manatus ◽  
Trichechus Manatus Latirostris ◽  
Potential Biomarker ◽  
Herpesvirus 1 ◽  
Quantitative Pcr Assay

Trichechid herpesvirus 1 (TrHV-1) is currently the only known herpesvirus in any sirenian. We hypothesized that stress may lead to recrudescence of TrHV-1 in manatees, thus making TrHV-1 a potential biomarker of stress. We optimized and validated a TrHV-1 real-time quantitative probe hybridization PCR (qPCR) assay that was used to quantify TrHV-1 in manatee peripheral blood mononuclear cells (PBMCs). Average baseline TrHV-1 loads in a clinically healthy wild Florida manatee ( Trichechus manatus latirostris) population ( n = 42) were 40.9 ± SD 21.2 copies/100 ng DNA; 19 of 42 manatees were positive. TrHV-1 loads were significantly different between the 2 field seasons ( p < 0.025). This optimized and validated qPCR assay may be used as a tool for further research into TrHV-1 in Florida manatees.

Sign in / Sign up

Export Citation Format

Share Document