Anti-MAdCAM Antibody Increases ß7+ T Cells and CCR9 Gene Expression in the Peripheral Blood of Patients With Crohn's Disease

Mina Hassan-Zahraee, Anindita Banerjee, John B Cheng, Weidong Zhang, Alaa Ahmad, Karen Page, David von Schack, Baohong Zhang, Steven W Martin, Satyaprakash Nayak, Padma Reddy, Li Xi, Hendrik Neubert, Mireia Fernandez Ocana, Ken Gorelick, Robert Clare, Michael Vincent, Fabio Cataldi, Kenneth Hung, Mina Hassan-Zahraee, Anindita Banerjee, John B Cheng, Weidong Zhang, Alaa Ahmad, Karen Page, David von Schack, Baohong Zhang, Steven W Martin, Satyaprakash Nayak, Padma Reddy, Li Xi, Hendrik Neubert, Mireia Fernandez Ocana, Ken Gorelick, Robert Clare, Michael Vincent, Fabio Cataldi, Kenneth Hung

Abstract

Objective: To define pharmacodynamic biomarkers in the peripheral blood of patients with Crohn's disease [CD] after treatment with PF-00547659, an anti-human mucosal addressin cell adhesion molecule-1 [MAdCAM-1] monoclonal antibody.

Methods: In this Phase 2, randomised, double-blind, controlled study [OPERA], blood samples were analysed from patients with moderate to severe active CD who received placebo or 22.5 mg, 75 mg, or 225 mg of PF-00547659 subcutaneously at baseline and at Weeks 4 and 8, with follow-up at Week 12. Soluble MAdCAM [sMAdCAM] was measured by mass spectrometry, β7-expressing T cells by flow cytometry, and gene transcriptome by RNA sequencing.

Results: A slight increase in sMAdCAM was measured in the placebo group from baseline to Week 12 [6%], compared with significant decreases in all PF-00547659 groups [-87% to -98%]. A slight increase from baseline to Week 12 was observed in frequency and molecules of equivalent soluble fluorochrome for β7+ central memory T cells in the placebo group [4%], versus statistically significant increases in the active treatment groups [48% to 81%]. Similar trends were seen for β7+ effector memory T cells [placebo, 8%; PF-00547659, 84-138%] and β7+ naïve T cells [8%; 13-50%]. CCR9 gene expression had statistically significant up-regulation [p = 1.09e-06; false discovery rate < 0.1] with PF-00547659 treatment, and was associated with an increase in β7+ T cells.

Conclusions: Results of the OPERA study demonstrate positive pharmacology and dose-dependent changes in pharmacodynamic biomarker measurements in blood, including changes in cellular composition of lymphocytes and corresponding CCR9 gene expression changes.

Trial registration: ClinicalTrials.gov NCT01276509.

Keywords: Crohn’s disease; MAdCAM; PF-00547659; pharmacodynamics; treatment.

Copyright © 2017 European Crohn’s and Colitis Organisation (ECCO). Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oup.com

Figures

Figure 1.
Figure 1.
The time course of serum soluble MAdCAM levels [geometric mean 90% CI, ng/ml] in patients following treatment with placebo or PF-00547659. Different symbols represent different dose groups: ○ placebo, ▲ PF-00547659 22.5 mg, ■ PF-00547659 75 mg, and ● PF-00547659 225 mg. CI, confidence interval; sMAdCAM, soluble mucosal addressin cell adhesion molecule.
Figure 2.
Figure 2.
Effect of PF-00547659 on [A] β7+ central memory T cells; [B] β7+ effector memory T cells; and [C] β7+ naïve T cells, in CD4+ cell subsets of patients with CD. β7 expression was measured by a validated whole blood assay [see Supplementary Figure 1 for gating strategy]. The error bars are geometric mean estimates [90% CI] for percent change from baseline in MESF. CI, confidence interval; MESF, molecules of equivalent soluble fluorochrome.
Figure 3.
Figure 3.
Effect of PF-00547659 on percent change from baseline of β7 expression in CD4+ cell subsets of patients with CD in the 225-mg group. Geometric mean % MESF change from baseline on β7+ in central memory, effector memory, and naïve T cells are calculated from data in Figure 2. CD, Crohn’s disease; FACS, fluorescence-activated cell sorting; MESF, molecules of equivalent soluble fluorochrome.
Figure 4.
Figure 4.
Fold changes in CCR9 gene expression [measured by counts per million] from baseline to Week 12 by treatment group. Each vertical line represents the 90% confidence interval for each fold change.
Figure 5.
Figure 5.
Expression profile of TReg-cell–related genes across treatment arms, ie in placebo, and PF-00547659 22.5-mg, 75-mg, and 225-mg groups, at Week 12. TReg, T regulatory cells.
Figure 6.
Figure 6.
Canonical pathways enriched in each treatment group with a z-score ≥ 2 or ≤ –2 and –log [p-value] > 1.3. Enriched pathways with z-scores of ≥ 2 were activated by differentially expressed genes, whereas those with z-scores ≤ –2 indicated inhibition of the enriched pathways. All enriched pathways met the statistical significance cut-off of p < 0.05.

References

    1. Pullen N, Molloy E, Carter D et al. . Pharmacological characterization of PF-00547659, an anti-human MAdCAM monoclonal antibody. Br J Pharmacol 2009;157:281–93.
    1. Vermeire S, Sandborn WJ, Danese S et al. . Anti-MAdCAM antibody [PF-00547659] for ulcerative colitis [TURANDOT]: a phase 2, randomised, double-blind, placebo-controlled trial. Lancet 2017, May 17. doi: 10.1016/S0140-6736[17]30930–3. [Epub ahead of print.]
    1. Sandborn WJ, Lee S, Tarabar D et al. . A Phase 2 evaluation of anti-MAdCAM antibody PF-00547659 in the treatment of Crohn’s disease: Report of the OPERA study. Gut 2017. [in press].
    1. Palandra J, Finelli A, Zhu M, Masferrer J, Neubert H. Highly specific and sensitive measurements of human and monkey interleukin 21 using sequential protein and tryptic peptide immunoaffinity LC-MS/MS. Anal Chem 2013;85:5522–9.
    1. Neubert H, Muirhead D, Kabir M, Grace C, Cleton A, Arends R. Sequential protein and peptide immunoaffinity capture for mass spectrometry-based quantification of total human β-nerve growth factor. Anal Chem 2013;85:1719–26.
    1. Sun H, Liu J, Zheng Y, Pan Y, Zhang K, Chen J. Distinct chemokine signaling regulates integrin ligand specificity to dictate tissue-specific lymphocyte homing. Dev Cell 2014;30:61–70.
    1. Singh H, Grewal N, Arora E, Kumar H, Kakkar AK. Vedolizumab: A novel anti-integrin drug for treatment of inflammatory bowel disease. J Nat Sci Biol Med 2016;7:4–9.
    1. Dobin A, Davis CA, Schlesinger F et al. . STAR: ultrafast universal RNA-seq aligner. Bioinformatics 2013;29:15–21.
    1. Quinlan AR, Hall IM. BEDTools: a flexible suite of utilities for comparing genomic features. Bioinformatics 2010;26:841–2.
    1. Robinson MD, McCarthy DJ, Smyth GK. edgeR: a bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics 2010;26:139–40.
    1. Chindelevitch L, Ziemek D, Enayetallah A et al. . Causal reasoning on biological networks: interpreting transcriptional changes. Bioinformatics 2012;28:1114–21.
    1. Dalmasso C. LBE: Estimation of the False Discovery Rate. R package version 1.26.0. 2007 Accessed December 9, 2015.
    1. Subramanian A, Tamayo P, Mootha VK et al. . Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc Natl Acad Sci U S A 2005;102:15545–50.
    1. Rodriguez MW, Paquet AC, Yang YH, Erle DJ. Differential gene expression by integrin beta 7+ and beta 7- memory T helper cells. BMC Immunol 2004;5:13.
    1. Gorfu G, Rivera-Nieves J, Ley K. Role of beta7 integrins in intestinal lymphocyte homing and retention. Curr Mol Med 2009;9:836–50.
    1. Vermeire S, Sandborn WJ, Danese S et al. . Anti-MAdCAM antibody [PF-00547659] for ulcerative colitis: the TURANDOT study. Lancet 2017;390:135–44.

Source: PubMed

Спонсоры и соавторы

Заболевания

Медикаментозные вмешательства

Подписаться