Genomic Responses of Human Immune and Non-Immune Cells to Glucocorticoids

Normal Physiology Clinical Trial

Official title:

Genomic Response of Human Immune and Non-Immune Cells to Glucocorticoids

Clinical Trial Details — Status: Completed

Administrative data

• NCT number: NCT02798523
• Other study ID #: 160126
• Secondary ID: 16-I-0126
• Status: Completed
• Phase: Phase 1
• Start date: January 24, 2017
• Est. completion date: November 14, 2022

Study information

• Verified date: November 14, 2022
• Source: National Institutes of Health Clinical Center (CC)
• Contact: n/a
• Is FDA regulated: No
• Study type: Interventional

Clinical Trial 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.

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.

Recruitment information / eligibility

• Status: Completed
• Enrollment: 33
• Est. completion date: November 14, 2022
• Est. primary completion date: May 31, 2022
• Accepts healthy volunteers: No
• Gender: All
• Age group: 18 Years to 64 Years

Eligibility

• INCLUSION CRITERIA:

1. Age 18 to 64 years

2. Willingness to have samples stored for future research

3. Willingness to undergo genetic testing

EXCLUSION CRITERIA:

1. Body Mass Index less than 18 or greater than 35

2. Difficult peripheral venous access (as determined by study staff at screening)

3. History of severe allergic reaction to glucocorticoids

4. History of autoimmune or autoinflammatory disease

5. Active solid or hematologic malignancy

6. History of a skin condition (such as psoriasis, pemphigus, or atopic dermatitis) that could affect the results of the transcriptional analysis of the skin biopsy samples

7. Diabetes mellitus

8. Cancer chemotherapy within the past 5 years

9. Surgery within the past 8 weeks

10. History of recent (within the past 30 days) infection

11. A positive test for human immunodeficiency virus, or hepatitis A, B or C virus infection (viral markers hepatitis screen, which includes HBsAg, anti-HCV IgG, anti-HAV IgM).

12. A positive or indeterminate test for latent tuberculosis (interferon gamma release assay)

13. History of parasitic, amebic, fungal or mycobacterial infections, or other possible latent infections

14. Coagulation test (PT and PTT) results outside of normal range

15. History of a bleeding disorder

16. Use of a glucocorticoid (including topical or inhaled), a nonsteroidal anti-inflammatory drug (including aspirin), an anti-epileptic drug, an anticoagulant, a statin, fluoxetine, diltiazem, amiodarone, clarithromycin, ketoconazole, or St. John s wort, within the past 30 days

17. Vaccination within the past 30 days

18. Receipt of an immunosuppressant or immunomodulatory drug within the past 30 days

19. Pregnancy, current or within the past 90 days, or trying to become pregnant during the study

20. Current breastfeeding

21. Complete blood count (CBC) and/or acute care panel values are both outside of the NIH Department of Laboratory Medicine normal reference range and deemed clinically significant by the principal investigator

22. Any Electrocardiogram (ECG) abnormality that is clinically significant

23. Any condition that, in the investigator s opinion, may put the participant at undue risk or compromise the study s scientific objectives

24. Participation in a clinical protocol which includes an intervention that, in the opinion of the investigator, may affect the results of the current study

Related Conditions & MeSH terms

• Normal Physiology

Intervention

Drug:
• Methylprednisolone sodium succinate(Solu-Medrol)
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.
• Topical methylprednisolone (Advantan emulsion 0 /1%)
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.

Locations

Country	Name	City	State
United States	National Institutes of Health Clinical Center	Bethesda	Maryland

Sponsors (1)

Lead Sponsor	Collaborator
National Institute of Allergy and Infectious Diseases (NIAID)

Country where clinical trial is conducted

United States, 

References & Publications (3)

Franco LM, Gadkari M, Howe KN, Sun J, Kardava L, Kumar P, Kumari S, Hu Z, Fraser IDC, Moir S, Tsang JS, Germain RN. Immune regulation by glucocorticoids can be linked to cell type-dependent transcriptional responses. J Exp Med. 2019 Feb 4;216(2):384-406. doi: 10.1084/jem.20180595. Epub 2019 Jan 23. — View Citation

Gadkari M, Makiya MA, Legrand F, Stokes K, Brown T, Howe K, Khoury P, Hu Z, Klion A, Franco LM. Transcript- and protein-level analyses of the response of human eosinophils to glucocorticoids. Sci Data. 2018 Dec 4;5:180275. doi: 10.1038/sdata.2018.275. — View Citation

Khoury P, Stokes K, Gadkari M, Makiya MA, Legrand F, Hu Z, Klion A, Franco LM. Glucocorticoid-induced eosinopenia in humans can be linked to early transcriptional events. Allergy. 2018 Oct;73(10):2076-2079. doi: 10.1111/all.13497. Epub 2018 Jul 17. No abstract available. — View Citation

Outcome

Type	Measure	Description	Time frame	Safety issue
Primary	Number of Participants Sampled for RNA-seq Differential Expression Analysis (Biological Replicates)	Number of participants (biological replicates) that were successfully sampled for RNA-seq differential gene expression analysis in glucocorticoid-treated immune cells. The analysis employed a cutoff value of < 5% false-discovery rate (FDR) to select the transcripts that were considered differentially expressed at each time point. The resulting gene lists were contrasted to determine which genes were uniquely differentially expressed in different cell types.	Up to 2 or 4 hours post infusion depending on group

External Links

•   NIH Clinical Center Detailed Web Page