Ex Vivo Induction of mRNA in Human Whole Blood as a New Platform of Drug and Dietary Supplement Development

PurposeWe introduced a new concept of ex vivo gene expression analysis (Mitsuhashi, Clin Chem 53:148–149, 2007), where drug action was simulated under physiological conditions. This model system was applied to study various fields of drug development.Materials and MethodsHeparinized human whole blood was incubated with drugs for less than 4h. The changes of specific mRNA were then quantified using the method we developed (Mitsuhashi, Tomozawa, Endo, and Shinagawa, Clin Chem 52:634–642, 2006).ResultsThe mRNA quantitation method was used as a model system to study the following areas: (1) identification of respondents and non-respondents, (2) ex vivo compound screening, (3) determination of individually optimized doses, (4) drug-to-drug comparison, (5) assessment of leukocyte toxicity, (6) discovery of molecular targets, (7) assessment of the action of dietary supplements, and (8) characterization of respondents and non-respondents for various dietary supplements.ConclusionSince ex vivo assays are safe, a large number of healthy donors and disease patients can be recruited to identify individual-to-individual variations, which is not available from current preclinical study models. Although each system should be validated using a large number of samples, the ex vivo analysis will be a new tool for the development of drugs and dietary supplements in future.

• Publication year

  2008

• Venue

  Pharmaceutical Research

• Publication date

  2008-01-09

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1007/s11095-007-9510-2PMID 18183479PMCID 2292497
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Ex vivo simulation of drug action: quantification of drug-induced mRNA as a bridge between preclinical and clinical trials.

  2007influential reference
• Prolactin enhances interferon-gamma-induced production of CXC ligand 9 (CXCL9), CXCL10, and CXCL11 in human keratinocytes.

  2007cited by this paper
• Quantification of mRNA in whole blood by assessing recovery of RNA and efficiency of cDNA synthesis.

  2006influential reference
• Sensitivity of whole-blood T lymphocytes in individual patients to tacrolimus (FK 506): impact of interleukin-2 mRNA expression as surrogate measure of immunosuppressive effect.

  2004cited by this paper
• Monitoring of NFAT-regulated gene expression in the peripheral blood of allograft recipients: a novel perspective toward individually optimized drug doses of cyclosporine A

  2004cited by this paper
• IgG Fc receptor polymorphisms in human disease: implications for intravenous immunoglobulin therapy.

  2003cited by this paper
• Differential chemokine expression profiles in human peripheral blood T lymphocytes: dependence on T-cell coreceptor and calcineurin signaling.

  2003cited by this paper
• No association between lck gene polymorphisms and protein level in type 1 diabetes.

  2002cited by this paper
• PUMA induces the rapid apoptosis of colorectal cancer cells.

  2001cited by this paper
• Expression of bbc3, a pro-apoptotic BH3-only gene, is regulated by diverse cell death and survival signals

  2001cited by this paper
• PUMA, a novel proapoptotic gene, is induced by p53.

  2001cited by this paper
• Toll-like Receptor-4 Mediates Lipopolysaccharide-induced Signal Transduction*

  1999cited by this paper
• Quantification of low-copy transcripts by continuous SYBR Green I monitoring during amplification.

  1998cited by this paper
• The Phosphotyrosine Phosphatase SHP-2 Participates in a Multimeric Signaling Complex and Regulates T Cell Receptor (TCR) coupling to the Ras/Mitogen-activated Protein Kinase (MAPK) Pathway in Jurkat T Cells

  1998cited by this paper
• Toll-like receptor-2 mediates lipopolysaccharide-induced cellular signalling

  1998cited by this paper
• Direct reverse transcription-PCR on oligo(dT)-immobilized polypropylene microplates after capturing total mRNA from crude cell lysates.

  1998influential reference
• Software to determine optimal oligonucleotide sequences based on hybridization simulation data.

  1996cited by this paper
• The EMT/ITK/TSK (EMT) tyrosine kinase is activated during TCR signaling: LCK is required for optimal activation of EMT.

  1996cited by this paper
• Oligonucleotide probe design — a new approach

  1994cited by this paper
• Cell cycle. p21 inhibits cyclin shock.

  1994cited by this paper
• Differential regulation of lymphotoxin (LT), lymphotoxin-beta (LT-beta), and TNF-alpha in murine T cell clones activated through the TCR.

  1994cited by this paper
• p21 inhibits cyclin shock

  1994cited by this paper
• Gene manipulation on plastic plates

  1992influential reference
• Dinucleotide repeat polymorphism at the human MHC class I CD8 alpha-chain (Leu-2/T8) gene.

  1991cited by this paper
• Detection of specific polymerase chain reaction product by utilizing the 5'----3' exonuclease activity of Thermus aquaticus DNA polymerase.

  1991cited by this paper
• Human T-cell receptor CD3-gamma (CD3G)/MspI DNA polymorphism.

  1989cited by this paper
• A cytosolic binding protein for the immunosuppressant FK506 has peptidyl-prolyl isomerase activity but is distinct from cyclophilin

  1989influential reference
• Human T-cell receptor CD3-epsilon (CD3E)/TaqI DNA polymorphism.

  1989cited by this paper
• Human T-cell receptor CD3-delta (CD3D)/MspI DNA polymorphism.

  1989cited by this paper
• Human T-cell receptor CD3- ε (CD3D) / TaqI DNA polymorphism

  1989cited by this paper
• Human T-cell receptor CD3-d (CD3D) / MspI DNA polymorphism

  1989cited by this paper
• Fc-receptor cross-linking induces rapid secretion of tumor necrosis factor (cachectin) by human peripheral blood monocytes.

  1988cited by this paper
• Inhibition of cytokine production by cyclosporin A and transforming growth factor beta

  1987cited by this paper
• Production and characterization of heat-aggregated IgG complexes with pre-determined molecular masses: light-scattering study.

  1987cited by this paper
• Cyclophilin: a specific cytosolic binding protein for cyclosporin A.

  1984cited by this paper

• Rapid and high throughput assessment of cellular immunity against SARS-CoV-2 based on the ex vivo activation of genes in leukocyte assay with whole blood.

  2023cites this paper
• Combinative effects of thanh hao miet giap thang (sweet wormwood and tortoise shell decoction) ingredients on antioxidative activity in vitro.

  2014cites this paper
• Ex vivo TCR‐induced leukocyte gene expression of inflammatory mediators is increased in type 1 diabetic patients but not in overweight children

  2010cites this paper
• Ex vivo simulation of leukocyte function: stimulation of specific subset of leukocytes in whole blood followed by the measurement of function-associated mRNAs.

  2010cites this paper
• A dried tofu-supplemented diet affects mRNA expression of inflammatory cytokines in human blood.

  2010cites this paper
• cDNA targets improve whole blood gene expression profiling and enhance detection of pharmocodynamic biomarkers: a quantitative platform analysis

  2010cites this paper
• Trastuzumab-induced CCL20 and interleukin-8 mRNA in human whole blood ex vivo

  2009cites this paper
• Enhanced Expression of Radiation-induced Leukocyte CDKN1A mRNA in Multiple Primary Breast Cancer Patients: Potential New Marker of Cancer Susceptibility

  2009cites this paper