Genomic Responses of Human Immune and Non-Immune Cells to Glucocorticoids
Condition: Normal Physiology
Study type: Interventional
Phase: Phase 1

Summary: 
      Background:

      The immune system defends the body against bacteria and other harmful invaders. But it can
      overact and attack healthy cells by mistake. The group of drugs called glucocorticoids (GCs)
      can calm down an overactive immune system. But they often cause negative side effects.
      Researchers want to learn how human genes respond to GCs. Genes live inside each cell of the
      body. They tell our cells how to function. Researchers hope the results of this study will
      show them how to develop better drugs that will have the benefits of GCs without the side
      effects.

      Objectives:

      To study how human genes respond to glucocorticoid drugs.

      Eligibility:

      Healthy adult volunteers ages 18-64.

      Design:

      Participants will be screened with a medical history and physical exam. They will have a
      heart test and blood tests.

      The study visit will last about 6 hours.

      Participants will have medical history, physical exam, and 3 blood draws.

      Participants will have a skin biopsy. An injection will numb the skin on one arm. Then a tool
      will remove a piece of skin about as big as a pencil eraser.

      A GC cream will be applied to the other arm.

      Participants will get the GC study drug for 30 minutes. It will be a liquid that will drip
      through a needle placed in an arm vein.

      Participants will have a skin biopsy of the arm that had the cream applied.

      Participants will have follow-up calls 1 and 4 days later. They will be asked about reactions
      or other health problems.
    

Detailed description: 
      Glucocorticoids are among the most frequently prescribed immunosuppressive and
      anti-inflammatory medications worldwide. Long-term use, however, is complicated by serious
      non-immunologic side effects. Ongoing in vitro experiments with human primary cells in our
      laboratory suggest that there are indeed fundamental differences in the genomic response of
      immune and non-immune cells to glucocorticoids. These and other aspects of drug action at the
      genomic level have not been completely characterized. This study will attempt to generate a
      list of human genes and non-coding RNAs that are differentially expressed and regulated in
      response to glucocorticoids between immune and non-immune cells. These data will be used to
      identify transcripts, their corresponding proteins, and the molecular pathways that are best
      candidates for targeted intervention. Potential targets could be validated with small
      interfering RNA (siRNA) libraries, with the long-term goal of developing small-molecule or
      nanoparticle-facilitated RNA interference (RNAi) interventions that reproduce the therapeutic
      action of glucocorticoids in immune cells while avoiding their harmful side effects on other
      tissues.

      Healthy volunteers will undergo baseline blood collection prior to receiving a single
      intravenous dose of 250 milligrams of methylprednisolone sodium succinate. Blood will be
      collected in one of two regimens: 1 and 2 hours or 2 and 4 hours after the start of the
      infusion. A skin punch biopsy may be obtained before healthy volunteers receive IV
      methylprednisolone. If so, topical methylprednisolone will be applied to a limited area of
      skin, contralateral to the site of the baseline skin biopsy, and an additional skin biopsy
      will be obtained 4 hours after drug administration, from the area where topical
      methylprednisolone was applied. Follow-up phone calls 1 day and 1 week after discharge will
      document any adverse effects related to the drug or skin biopsy. Total length of individual
      study participation is 1-5 weeks.

      Blood samples will be processed for isolation of hematopoietic cell sub-population
      (neutrophils, B cells, CD4+ T cells, CD8+ T cells, monocytes, and natural killer [NK] cells).
      Laboratory studies will be performed in the purified cells, with the goal of understanding
      the human response to glucocorticoids in vivo at the level of RNA (e.g., RNA sequencing,
      small -RNA-sequencing, real-time PCR), DNA (e.g., ChIP-seq, methylation analysis, DNA
      sequencing, genotyping, and protein (e.g., flow cytometry, mass spectrometry). At each time
      point, serum methylprednisolone levels will be measured and flow cytometry for standard
      lineage markers will be performed. Skin biopsies will be subjected to RNA isolation for RNA
      sequencing and small-RNA sequencing. A fragment of each skin biopsy will undergo fibroblast
      isolation and culture for in vitro exposure to glucocorticoids and for the generation of
      induced pluripotent stem (iPS) cells.
    

Intervention name(s): Methylprednisolone sodium succinate(Solu-Medrol); Topical methylprednisolone (Advantan emulsion 0 /1%)
Intervention type(s): Drug; Drug
Intervention description(s): Methylprednisolone sodium succinate for injection, USP (SOLU-MEDROL sterile powder, Pfizer, Inc.) is an anti-inflammatory glucocorticoid which occurs as a white, or nearly white, odorless hygroscopic, amorphous solid. It is very soluble in water and in alcohol; it is insoluble in chloroform and is very slightly soluble in acetone.
125-milligram Act-O-Vial (AOV) System: Each 2 mL AOV (when mixed) contains methylprednisolone sodium succinate equivalent to 125 milligrams methylprednisolone; also 1.6 mg monobasic sodium phosphate anhydrous; and 17.4 mg dibasic sodium phosphate dried.; 1 g Advantan emulsion 0.1% (Bayer) contains methylprednisolone aceponate (21-acetoxy-11beta-hydroxy-6alpha-methyl-17-propionyloxy-1,4-pregnadiene-3,20-dione) 1 mg, as the active ingredient. It is an oil-in-water emulsion containing medium chain triglycerides, caprylic-capric-stearic triglyceride, polyoxyethylene alcohol 2-stearyl ether, polyoxyethylene alcohol-21-stearyl ether, benzyl alcohol, disodium edetate, glycerol, and purified water.
Intervention MeSH terms: Methylprednisolone; Methylprednisolone Acetate; Methylprednisolone Hemisuccinate; Prednisolone; Prednisolone acetate; Methylprednisolone aceponate; Prednisolone hemisuccinate; Prednisolone phosphate
Allocation: Non-Randomized
Intervention model: Sequential Assignment
Masking: None (Open Label)
Primary purpose: Basic Science
Keywords: Methylprednisolone; Immunologic; Glucocorticoids; Functional genomics; RNA; Steroids