Pluripotent stem cells (ES and iPS cells) have the ability to self-renew and to differentiate into multiple lineages in vitro. This makes these cells powerful tool to study early embryonic developmental pathways and to generate specific cell populations for regenerative medicine and disease investigation. Supported by R01 AR055299, our research group has pioneered methods to derive skeletal myogenic cells from mouse and human pluripotent ES and iPS cells. Until our work, studies in this area were rare since skeletal myogenic differentiation is extremely inefficient during in vitro differentiation. We reasoned that because in vitro systems do not recapitulate neural tube or notochord development, and indeed do not produce structures resembling somites, that the inducing signals that properly pattern paraxial mesoderm were absent. We have overcome this roadblock by activating the myogenic program through transient induction of Pax3 or Pax7 during early mesoderm development. This approach allows for the in vitro generation of large quantities of early embryonic skeletal myogenic progenitors. Transplantation of these cells into dystrophic mice results in myofiber and satellite cell engraftment that is accompanied by improvement in muscle force generation. This renewal application builds on the advances we have brought to the field over the past 5 years and evolves from these into 3 areas: i) understanding the molecular regulation of the embryonic myogenic program by Pax3, ii) investigating the long-term regenerative capacity of human pluripotent donor-derived satellite cells as well as their transcriptional profiling in comparison to in vitro generated myogenic progenitors, and iii) using myotubes from patient-specific pluripotent cells to biochemically and functionally model DMD/BMD pathology and its reversal by therapeutic delivery of dystrophin mini-genes.

Public Health Relevance

This proposal takes advantage of the ability of pluripotent stem cells to uniquely produce differentiated somatic cell types in large quantities. This allows the study of embryonic developmental pathways using biochemical methods, the generation of specific cell populations for regenerative medicine, and the modeling of disease and genetic correction in vitro. Although studying muscle development using pluripotent cell systems has historically been difficult or impossible because of the inefficiency with which pluripotent cells undergo skeletal muscle differentiation, we have developed methods, based on the conditional expression of myogenic master regulators, Pax3 and Pax7, to induce the muscle program in early mesoderm, and thus derive embryonic muscle progenitors with great efficiency. Our investigations will focus on 1) understanding the mechanism by which Pax3 regulates embryonic myogenesis;2) investigating the regenerative potential of donor-derived satellite cells and their molecular signature;3) in vitro disease modeling of DMD/BMD through the use of disease-specific iPS cell-derived myotubes.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
2R01AR055299-06A1
Application #
8814038
Study Section
Skeletal Muscle and Exercise Physiology Study Section (SMEP)
Program Officer
Boyce, Amanda T
Project Start
2007-07-01
Project End
2019-07-31
Budget Start
2014-09-11
Budget End
2015-07-31
Support Year
6
Fiscal Year
2014
Total Cost
$334,400
Indirect Cost
$114,400
Name
University of Minnesota Twin Cities
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
McCullagh, Karl J A; Perlingeiro, Rita C R (2015) Coaxing stem cells for skeletal muscle repair. Adv Drug Deliv Rev 84:198-207
Dandapat, Abhijit; Bosnakovski, Darko; Hartweck, Lynn M et al. (2014) Dominant lethal pathologies in male mice engineered to contain an X-linked DUX4 transgene. Cell Rep 8:1484-96
Magli, Alessandro; Schnettler, Erin; Swanson, Scott A et al. (2014) Pax3 and Tbx5 specify whether PDGFR?+ cells assume skeletal or cardiac muscle fate in differentiating embryonic stem cells. Stem Cells 32:2072-83
Rinaldi, Fabrizio; Perlingeiro, Rita C R (2014) Stem cells for skeletal muscle regeneration: therapeutic potential and roadblocks. Transl Res 163:409-17
Skoglund, Gunnar; Lainé, Jeanne; Darabi, Radbod et al. (2014) Physiological and ultrastructural features of human induced pluripotent and embryonic stem cell-derived skeletal myocytes in vitro. Proc Natl Acad Sci U S A 111:8275-80
Magli, Alessandro; Schnettler, Erin; Rinaldi, Fabrizio et al. (2013) Functional dissection of Pax3 in paraxial mesoderm development and myogenesis. Stem Cells 31:59-70
Parker, Sarah L; Perlingeiro, Rita C R (2013) Are we there yet? Navigating roadblocks to induced pluripotent stem cell therapy translation. Regen Med 8:389-91
Filareto, Antonio; Parker, Sarah; Darabi, Radbod et al. (2013) An ex vivo gene therapy approach to treat muscular dystrophy using inducible pluripotent stem cells. Nat Commun 4:1549
Darabi, Radbod; Santos, Filipe N C; Filareto, Antonio et al. (2011) Assessment of the myogenic stem cell compartment following transplantation of Pax3/Pax7-induced embryonic stem cell-derived progenitors. Stem Cells 29:777-90
Darabi, Radbod; Pan, Weihong; Bosnakovski, Darko et al. (2011) Functional myogenic engraftment from mouse iPS cells. Stem Cell Rev 7:948-57

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