Biomarker-focused multi-drug combination therapy and repurposing trial in mdx mice

PLoS One. 2021 Feb 22;16(2):e0246507. doi: 10.1371/journal.pone.0246507. eCollection 2021.


Duchenne muscular dystrophy is initiated by dystrophin deficiency, but downstream pathophysiological pathways such as membrane instability, NFĸB activation, mitochondrial dysfunction, and induction of TGFβ fibrosis pathways are thought to drive the disability. Dystrophin replacement strategies are hopeful for addressing upstream dystrophin deficiency; however, all methods to date use semi-functional dystrophin proteins that are likely to trigger downstream pathways. Thus, combination therapies that can target multiple downstream pathways are important in treating DMD, even for dystrophin-replacement strategies. We sought to define blood pharmacodynamic biomarkers of drug response in the mdx mouse model of Duchenne muscular dystrophy using a series of repurposed drugs. Four-week-old mdx mice were treated for four weeks with four different drugs singly and in combination: vehicle, prednisolone, vamorolone, rituximab, β-aminoisobutyric acid (BAIBA) (11 treatment groups; n = 6/group). Blood was collected via cardiac puncture at study termination, and proteomic profiling was carried out using SOMAscan aptamer panels (1,310 proteins assayed). Prednisolone was tested alone and in combination with other drugs. It was found to have a good concordance of prednisolone-responsive biomarkers (56 increased by prednisolone, 39 decreased) focused on NFκB and TGFβ cascades. Vamorolone shared 45 (80%) of increased biomarkers and 13 (33%) of decreased biomarkers with prednisolone. Comparison of published human corticosteroid-responsive biomarkers to our mdx data showed 14% (3/22) concordance between mouse and human. Rituximab showed fewer drug-associated biomarkers, with the most significant being human IgG. On the other hand, BAIBA treatment (high and low dose) showed a drug-associated increase in 40 serum proteins and decreased 5 serum proteins. Our results suggest that a biomarker approach could be employed for assessing drug combinations in both mouse and human studies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aminoisobutyric Acids / therapeutic use
  • Animals
  • Biomarkers
  • Disease Models, Animal
  • Drug Repositioning
  • Male
  • Mice
  • Mice, Inbred mdx
  • Muscular Dystrophy, Animal / drug therapy*
  • Muscular Dystrophy, Duchenne / drug therapy*
  • Prednisolone / therapeutic use
  • Pregnadienediols / therapeutic use
  • Rituximab / therapeutic use


  • Aminoisobutyric Acids
  • Biomarkers
  • Pregnadienediols
  • VBP15 compound
  • Rituximab
  • Prednisolone
  • 3-aminoisobutyric acid

Grant support

Funded by the Foundation to Eradicate Duchenne. The authors would like to thank the Foundation to Eradicate Duchenne for financial support of this study. The financers had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.