Chronic muscle diseases including Duchenne muscular dystrophy (DMD) and aging-related sarcopenia result in muscle weakness, loss of independence, and increased risk of death. In addition, traumatic muscle injury and loss due to accidents, surgery, and wartime injuries needs prolonged recovery. Skeletal muscle is a highly regenerative tissue in which satellite cells, a stem cell population for skeletal muscle, play essential roles in creating and repairing skeletal muscle. However, this potential ultimately fails with disease and aging. Autologous satellite cell transplantation is a potential approach to create and repair skeletal muscle fibers, but satellite cells are rare (a few % of all muscle nuclei) and often difficult to isolate. Patient-derived induced pluripotent stem cells (iPSCs) are the ideal cell source to obtain an unlimited number of myogenic cells that escape immune rejection after engraftment. However, efficient myogenic differentiation and the scale-up of myogenic differentiation remain elusive and must be developed further in order to generate effective cellular therapies. A key to the generation of human myogenic cells and skeletal muscle in a host animal is the selective knockout of genes in the blastocyst that are critical for organ development. Therefore, in this proposal, (1) we will determine to which extent mouse iPSC-derived limb skeletal muscle will be generated after injection of mouse iPSCs into Pax3 mutant mouse blastocysts, creating a niche in which stem cells can occupy and form skeletal muscle in the limb. This approach will provide evidence for the creation of entire skeletal muscle by mouse iPSCs in vivo. In addition, (2) we will generate a humanized skeletal muscle using Pax3 mutant mouse embryos via in utero injection of human iPSCs in combination with Pax3 mutant embryos and iPSCs. This concerted approach will help us to create iPSC-derived skeletal muscle and myogenic cells in vivo that can be transplanted into patients for a definitive cure of myopathic diseases and muscle injuries. In addition, the humanized skeletal muscle in mice will serve as an animal model to study the characteristics and regeneration characteristics of the human skeletal muscle diseases and responses to pharmacological agents and to provide a proof of concept for generating patient-derived cell and tissue sources for autologous muscle transplantation. .

Public Health Relevance

Lay summary The innovation in our approach is to create humanized skeletal muscle and myogenic stem cells in animals with high efficiency. The creation of humanized skeletal muscle, healthy copies of a patient?s own skeletal muscle and myogenic stem cells, would revolutionize the field of regenerative medicine and transplantation for patients suffering myopathic diseases and muscle injuries.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21AR070319-02
Application #
9323297
Study Section
Skeletal Muscle and Exercise Physiology Study Section (SMEP)
Program Officer
Boyce, Amanda T
Project Start
2016-08-01
Project End
2018-06-30
Budget Start
2017-07-01
Budget End
2018-06-30
Support Year
2
Fiscal Year
2017
Total Cost
Indirect Cost
Name
University of Minnesota Twin Cities
Department
Neurology
Type
Schools of Medicine
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Lowe, Matthew; Lage, Jacob; Paatela, Ellen et al. (2018) Cry2 Is Critical for Circadian Regulation of Myogenic Differentiation by Bclaf1-Mediated mRNA Stabilization of Cyclin D1 and Tmem176b. Cell Rep 22:2118-2132
Verma, Mayank; Asakura, Yoko; Murakonda, Bhavani Sai Rohit et al. (2018) Muscle Satellite Cell Cross-Talk with a Vascular Niche Maintains Quiescence via VEGF and Notch Signaling. Cell Stem Cell 23:530-543.e9
Mademtzoglou, Despoina; Asakura, Yoko; Borok, Matthew J et al. (2018) Cellular localization of the cell cycle inhibitor Cdkn1c controls growth arrest of adult skeletal muscle stem cells. Elife 7:
Mohan, Amrudha; Asakura, Atsushi (2017) CDK inhibitors for muscle stem cell differentiation and self-renewal. J Phys Fit Sports Med 6:65-74
Chowdhury, Neeladri; Asakura, Atsushi (2017) In Utero Stem Cell Transplantation: Potential Therapeutic Application for Muscle Diseases. Stem Cells Int 2017:3027520
Hron, Alexander J; Asakura, Atsushi (2017) An Examination of the Role of Transcriptional and Posttranscriptional Regulation in Rhabdomyosarcoma. Stem Cells Int 2017:2480375
Kodaka, Yusaku; Rabu, Gemachu; Asakura, Atsushi (2017) Skeletal Muscle Cell Induction from Pluripotent Stem Cells. Stem Cells Int 2017:1376151
Kodaka, Yusaku; Asakura, Yoko; Asakura, Atsushi (2017) Spin infection enables efficient gene delivery to muscle stem cells. Biotechniques 63:72-76
Wang, Chao; Liu, Weiyi; Nie, Yaohui et al. (2017) Loss of MyoD Promotes Fate Transdifferentiation of Myoblasts Into Brown Adipocytes. EBioMedicine 16:212-223