Myoscaffolds reveal laminin scarring is detrimental for stem cell function while sarcospan induces compensatory fibrosis

Kristen M Stearns-Reider; Michael R Hicks; Katherine G Hammond; Joseph C Reynolds; Alok Maity; Yerbol Z Kurmangaliyev; Jesse Chin; Adam Z Stieg; Nicholas A Geisse; Sophia Hohlbauch; Stefan Kaemmer; Lauren R Schmitt; Thanh T Pham; Ken Yamauchi; Bennett G Novitch; Roy Wollman; Kirk C Hansen; April D Pyle; Rachelle H Crosbie

doi:10.1038/s41536-023-00287-2

Myoscaffolds reveal laminin scarring is detrimental for stem cell function while sarcospan induces compensatory fibrosis

NPJ Regen Med. 2023 Mar 15;8(1):16. doi: 10.1038/s41536-023-00287-2.

Authors

Kristen M Stearns-Reider^{1

2}, Michael R Hicks^{3

4

5}, Katherine G Hammond¹, Joseph C Reynolds¹, Alok Maity^{1

6

7}, Yerbol Z Kurmangaliyev^{7

8}, Jesse Chin¹, Adam Z Stieg⁹, Nicholas A Geisse¹⁰, Sophia Hohlbauch¹¹, Stefan Kaemmer¹², Lauren R Schmitt¹³, Thanh T Pham¹³, Ken Yamauchi^{4

14}, Bennett G Novitch^{4

14

15

16}, Roy Wollman^{1

6

7}, Kirk C Hansen¹³, April D Pyle^{3

4

15}, Rachelle H Crosbie^{17

18

19

20}

Affiliations

¹ Department of Integrative Biology and Physiology, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
² Department of Orthopaedic Surgery, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
³ Department of Microbiology, Immunology, and Molecular Genetics, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
⁴ Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
⁵ Department of Physiology and Biophysics, University of California, Irvine, Irvine, CA, 92697, USA.
⁶ Department of Chemistry and Biochemistry, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
⁷ Institute for Quantitative and Computational Biology, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
⁸ Department of Biological Chemistry, HHMI, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
⁹ California NanoSystems Institute, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
¹⁰ Curi Bio, Seattle, WA, 98121, USA.
¹¹ Asylum Research, An Oxford Instruments Company, Santa Barbara, CA, 93117, USA.
¹² Park Systems, 3040 Olcott St, Santa Clara, CA, 95054, USA.
¹³ Department of Biochemistry and Molecular Genetics, University of Colorado, Denver, Aurora, CO, 80045, USA.
¹⁴ Department of Neurobiology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
¹⁵ Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA, 90095, USA.
¹⁶ Intellectual and Developmental Disabilities Research Center, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, 90095, USA.
¹⁷ Department of Integrative Biology and Physiology, University of California, Los Angeles, Los Angeles, CA, 90095, USA. rcrosbie@physci.ucla.edu.
¹⁸ Eli and Edythe Broad Center of Regenerative Medicine and Stem Cell Research, University of California, Los Angeles, Los Angeles, CA, 90095, USA. rcrosbie@physci.ucla.edu.
¹⁹ Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA, 90095, USA. rcrosbie@physci.ucla.edu.
²⁰ Department of Neurology, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, CA, 90095, USA. rcrosbie@physci.ucla.edu.

Abstract

We developed an on-slide decellularization approach to generate acellular extracellular matrix (ECM) myoscaffolds that can be repopulated with various cell types to interrogate cell-ECM interactions. Using this platform, we investigated whether fibrotic ECM scarring affected human skeletal muscle progenitor cell (SMPC) functions that are essential for myoregeneration. SMPCs exhibited robust adhesion, motility, and differentiation on healthy muscle-derived myoscaffolds. All SPMC interactions with fibrotic myoscaffolds from dystrophic muscle were severely blunted including reduced motility rate and migration. Furthermore, SMPCs were unable to remodel laminin dense fibrotic scars within diseased myoscaffolds. Proteomics and structural analysis revealed that excessive collagen deposition alone is not pathological, and can be compensatory, as revealed by overexpression of sarcospan and its associated ECM receptors in dystrophic muscle. Our in vivo data also supported that ECM remodeling is important for SMPC engraftment and that fibrotic scars may represent one barrier to efficient cell therapy.

Abstract

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