Nanotopography-responsive myotube alignment and orientation as a sensitive phenotypic biomarker for Duchenne Muscular Dystrophy

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11 Scopus citations

Abstract

Duchenne Muscular Dystrophy (DMD) is a fatal genetic disorder currently having no cure. Here we report that culture substrates patterned with nanogrooves and functionalized with Matrigel (or laminin) present an engineered cell microenvironment to allow myotubes derived from non-diseased, less-affected DMD, and severely-affected DMD human induced pluripotent stem cells (hiPSCs) to exhibit prominent differences in alignment and orientation, providing a sensitive phenotypic biomarker to potentially facilitate DMD drug development and early diagnosis. We discovered that myotubes differentiated from myogenic progenitors derived from non-diseased hiPSCs align nearly perpendicular to nanogrooves, a phenomenon not reported previously. We further found that myotubes derived from hiPSCs of a dystrophin-null DMD patient orient randomly, and those from hiPSCs of a patient carrying partially functional dystrophin align approximately 14° off the alignment direction of non-diseased myotubes. Substrates engineered with micron-scale grooves and/or cell adhesion molecules only interacting with integrins all guide parallel myotube alignment to grooves and lose the ability to distinguish different cell types. Disruption of the interaction between the Dystrophin-Associated-Protein-Complex (DAPC) and laminin by heparin or anti-α-dystroglycan antibody IIH6 disenables myotubes to align perpendicular to nanogrooves, suggesting that this phenotype is controlled by the DAPC-mediated cytoskeleton-extracellular matrix linkage.

Original languageEnglish (US)
Pages (from-to)54-66
Number of pages13
JournalBiomaterials
Volume183
DOIs
StatePublished - Nov 2018

Bibliographical note

Funding Information:
This work was supported by the National Science Foundation ( CAREER DMR-1151529 for W.S.), by the National Institutes of Arthritis and Musculoskeletal and Skin Diseases of the National Institutes of Health ( R01 AR055299 for R.C.R.P.), by the Minnesota Partnership for Biotechnology and Medical Genomics (for Y.A. and S.J.K.), and by Institute for Engineering in Medicine at the University of Minnesota . Portions of this work were carried out in the Minnesota Nano Center, which receives partial support from NSF through the National Nanotechnology Coordinated Infrastructure (NNCI) , and the Characterization Facility at the University of Minnesota, which has received capital equipment funding from NSF through the MRSEC program under Award No. DMR-1420013 . The cytogenetic analyses were performed in the Cytogenomics Shared Resource at the University of Minnesota with support from the comprehensive Masonic Cancer Center NIH Grant # P30 CA077598-09 . We thank James Kiley for assistance with generation of hPSC-derived myogenic progenitors, Yi Ren for assistance with cell sorting, Tony Whipple for assistance with nanofabrication, and Patrick Alford for the use of the confocal microscope. We thank Allison Siehr for proofreading the manuscript.

Keywords

  • Duchenne Muscular Dystrophy
  • Laminin
  • Matrigel
  • Myotube orientation
  • Nanotopography
  • Stem cells

How much support was provided by MRSEC?

  • Shared

Reporting period for MRSEC

  • Period 5

PubMed: MeSH publication types

  • Journal Article
  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

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