Development of Potential Pharmacodynamic and Diagnostic Markers for Anti-IFN-α Monoclonal Antibody Trials in Systemic Lupus Erythematosus

Abstract

To identify potential pharmacodynamic biomarkers to guide dose selection in clinical trials using anti-interferon-alpha (IFN-α) monoclonal antibody (mAb) therapy for systemic lupus erythematosus (SLE), we used an Affymetrix human genome array platform and identified 110 IFN-α/β-inducible transcripts significantly upregulated in whole blood (WB) of 41 SLE patients. The overexpression of these genes was confirmed prospectively in 54 additional SLE patients and allowed for the categorization of the SLE patients into groups of high, moderate, and weak overexpressers of IFN-α/β-inducible genes. This approach could potentially allow for an accurate assessment of drug target neutralization in early trials of anti-IFN-α mAb therapy for SLE. Furthermore, ex vivo stimulation of healthy donor peripheral blood mononuclear cells with SLE patient serum and subsequent neutralization with anti-IFN-α mAb or anti-IFN-α receptor mAb showed that anti-IFN-α mAb has comparable effects of neutralizing the overexpression of type I IFN-inducible genes as that of anti-IFNAR mAb. These results suggest that IFN-α, and not other members of type I IFN family in SLE patients, is mainly responsible for the induction of type I IFN-inducible genes in WB of SLE patients. Taken together, these data strengthen the view of IFN-α as a therapeutic target for SLE.

Relationship between disease activity and type 1 interferon- and other cytokine-inducible gene expression in blood in dermatomyositis and polymyositis

“…36 All 42 patients satisfied widely used descriptive criteria for DM or PM. 37 In all, 13 overexpressed type 1 IFNinducible genes (from the 100 most overexpressed genes; Supplementary 30,35 Because there are no generally accepted gene expression signatures for the other cytokines used in this study, we generated empirically defined 13-gene set signatures for them as follows: whole blood collected from four healthy donors was exposed to either GM-CSF, IL-10, IL-13, IL-1b or TNF-a (R&D Systems, Minneapolis, MN, USA) at concentrations of 3 Â EC 50 , vehicle (1 Â phosphate-buffered saline) control, cytokine þ control antibody (R347, 10 mg ml À1 ) or cytokine þ cytokine-specific neutralizing monoclonal antibody (R&D Systems, 10 mg ml À1 ), as described. 38 Microarray experiments performed on extracted RNA were used to identify for each cytokine the 13 most highly overexpressed (average fold change 42) and cytokine-specific antibody-neutralized uniquely annotated genes (Supplementary Table 3).…”

“…Type 1 IFN-inducible genes are well characterized, and we defined this gene signature empirically from patient used in the study following procedures similarly to previously described for SLE. 30,35 In short, transcript profiling data were generated from the first visit of the 24 DM or PM patients enrolled in the longitudinal study in combination with a second cohort of 18 DM or PM patients screened for enrollment in a clinical trial of anti-IFN-a therapy for myositis. 36 All 42 patients satisfied widely used descriptive criteria for DM or PM.…”

Shifting of Immune Responsiveness to House Dust Mite by Influenza A Infection: Genomic Insights

“…cDNA was generated using the Super Script III Reverse Transcriptase kit (Life Technologies, Carlsbad, CA), according to the manufacturer's instructions. Relative transcript expression assay was conducted, as described previously (13), using the Fluidigm Biomark system (Fluidigm, San Francisco, CA). BestKeeper (version 1) (14) was used to identify the stably expressed housekeeping gene to be used as an internal reference.…”

A fully human monoclonal antibody with novel binding epitope and excellent neutralizing activity to multiple human IFN-α subtypes: A candidate therapy for systemic lupus erythematosus

“…The investigation into the genetics of SLE 12 has found significant overexpression of mRNAs of almost all IFN-a subtypes in the WB of SLE patients, making it important that IFN-a-targeted therapeutic agents neutralize extensive IFN-a subtypes. Additionally, the possibility that treatment with IFNAR-targeted agents may provide greater neutralization than IFN-a-targeted strategies should be considered.…”