Prediction of combination therapies based on topological modeling of the immune signaling network in multiple sclerosis

Abstract

Background Multiple sclerosis (MS) is a major health problem, leading to a significant disability and patient suffering. Although chronic activation of the immune system is a hallmark of the disease, its pathogenesis is poorly understood, while current treatments only ameliorate the disease and may produce severe side effects. Methods Here, we applied a network-based modeling approach based on phosphoproteomic data to uncover the differential activation in signaling wiring between healthy donors, untreated patients, and those under different treatments. Based in the patient-specific networks, we aimed to create a new approach to identify drug combinations that revert signaling to a healthy-like state. We performed ex vivo multiplexed phosphoproteomic assays upon perturbations with multiple drugs and ligands in primary immune cells from 169 subjects (MS patients, n=129 and matched healthy controls, n=40). Patients were either untreated or treated with fingolimod, natalizumab, interferon-β, glatiramer acetate, or the experimental therapy epigallocatechin gallate (EGCG). We generated for each donor a dynamic logic model by fitting a bespoke literature-derived network of MS-related pathways to the perturbation data. Last, we developed an approach based on network topology to identify deregulated interactions whose activity could be reverted to a “healthy-like” status by combination therapy. The experimental autoimmune encephalomyelitis (EAE) mouse model of MS was used to validate the prediction of combination therapies. Results Analysis of the models uncovered features of healthy-, disease-, and drug-specific signaling networks. We predicted several combinations with approved MS drugs that could revert signaling to a healthy-like state. Specifically, TGF-β activated kinase 1 (TAK1) kinase, involved in Transforming growth factor β-1 proprotein (TGF-β), Toll-like receptor, B cell receptor, and response to inflammation pathways, was found to be highly deregulated and co-druggable with all MS drugs studied. One of these predicted combinations, fingolimod with a TAK1 inhibitor, was validated in an animal model of MS. Conclusions Our approach based on donor-specific signaling networks enables prediction of targets for combination therapy for MS and other complex diseases.

Multiscale networks in multiple sclerosis

“…Fingolimod treatment induced a decrease in total CD4+ T cells and mature and memory B cells and increases in CD4+, CD8+ T-regulatory and B-regulatory cells (24). Finally, the phosphoproteomic analysis was carried out by conducting ex-vivo assays in PBMCs and quantified using xMAP assays on the first 148 patients at baseline as described before (25, 27), showing higher levels of phosphorylated IKBA, JUN, KSGB1, MK03, RS6, STAT3 and STAT6 in MS patients compared to controls ( Methods, File S1 ).…”

“…Phosphoprotein levels were quantified using xMAP assays performed blindly at ProtAtOnce (Athens, Greece) as described previously (25, 27). We analyzed a set of kinases associated with MS (9) which provides an adequate signal to noise ratio and test-retest reproducibility: AKT1, AKTS1, CREB1, GSK3AB, HSPB1, IKBA, JUN, KS6B1, LCK, MK12, MK03/01, MK09, MP2K1, NRF2, P53, PGFRB, PTN11, RS6, SRC, STAT1, STAT3, STAT5, STAT6, TF65, WNK1.…”

“…We analyzed a set of kinases associated with MS (9) which provides an adequate signal to noise ratio and test-retest reproducibility: AKT1, AKTS1, CREB1, GSK3AB, HSPB1, IKBA, JUN, KS6B1, LCK, MK12, MK03/01, MK09, MP2K1, NRF2, P53, PGFRB, PTN11, RS6, SRC, STAT1, STAT3, STAT5, STAT6, TF65, WNK1. Phosphoprotein data was normalized after the measurements were taken as described elsewhere (27).…”

Proteomics in Multiple Sclerosis: The Perspective of the Clinician

“…The model found the TGF-β-activated kinase 1 (TAK1) pathway, an important player of inflammatory and immune pathways, to be activated under all DMT treatment. After, utilizing the EAE mice model, based on these findings, they were able to validate the potential efficacy of FNG + TAK1-inhibitor combination therapy [201].…”

Epigallocatechin Gallate: A Multifaceted Molecule for Neurological Disorders and Neurotropic Viral Infections

“…71 EGCG administration in mouse models of MS led to a decrease in TGF-β activated kinase 1 (TAK1), which is involved in transforming growth factor β-1 proprotein (TGF-β), toll-like receptor, and B-cell receptor in response to inflammatory pathways (Figure 1II). 72 Intraperitoneal injection of EGCG (50 mg/kg/daily) in MSmice was found to elevate the expression level of proteolipid protein (PLP) and oligodendrocyte transcription factor 1 (Olig1) in transcriptional levels compared to controls. 73 Treating PBMCs isolated from MS patients with EGCG (25, 50, and 100 μM) for 18 h showed a significant decrease in RORC2 gene levels which acts as a transcription factor in the process of evolution of Th17 cells.…”