In the most prevalent muscular dystrophy, Duchenne Muscular Dystrophy (DMD), repeated muscle degeneration and regeneration leads to muscle satellite cell (SC) dysfunction and/or exhaustion and there is no cure. In the previous funding period, we have identified novel differentiation and enrichment strategies to generate the most engraftable cells to date from human induced pluripotent stem cells (hiPSCs), as well as a CRISPR correction strategy to restore dystrophin applicable to 60% of patients. The next phase of this work is now focused on improving our understanding of the functional status of skeletal muscle progenitors (SMPCs) derived from wt, DMD and CRISPR corrected lines. This work will improve our understanding of the molecular and differences between human PAX7+ stem cells and progenitor cells across human fetal development through adulthood, and inform our ability to generate the most regenerative cells from hiPSCs in this funding period.
In Aim 1, we will define human SMPCs and SCs arising in development and from hiPSCs and identify functional differences between progenitor and SC states across human development and in vitro derived cells using single cell sequencing and evaluation of candidate pathways different between SMPC and SC states.
In Aim 2, we will evaluate the role of the host microenvironment including endogenous PAX7 cells on stem cell engraftment and ability to transition to SCs and reside in the SC niche.
In Aim 3, we will utilize DMD and isogenic CRISPR/Cas9 SMPCs/SCs to evaluate specification, cell biology and functional potential of DMD and CRISPR corrected cells in diseased mdx-NSG and mdx-D2-NSG microenvironments. This will improve our understanding of differences and transitions between human muscle progenitor and stem cell states and will improve our ability to generate cells capable of repopulating the niche in long term studies.

Public Health Relevance

Skeletal muscle progenitor cells (SMPCs) and satellite cells (SCs) participate in muscle growth and regeneration in fetal and adult development, respectively. In human, the transition of SMPCs to SCs during development is poorly understood. Human induced pluripotent stem cells (hiPSCs) offer enormous potential for understanding human myogenesis including the transition of progenitors to adult SCs. Generation of the most regenerative SMPC or SC-like cells from hiPSCs could also improve our ability to model muscle diseases in a dish and have the potential for use in personalized cell replacement therapies.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
2R01AR064327-06A1
Application #
9662332
Study Section
Skeletal Muscle Biology and Exercise Physiology Study Section (SMEP)
Program Officer
Cheever, Thomas
Project Start
2013-09-20
Project End
2023-11-30
Budget Start
2019-01-01
Budget End
2019-11-30
Support Year
6
Fiscal Year
2019
Total Cost
Indirect Cost
Name
University of California Los Angeles
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Hicks, Michael R; Hiserodt, Julia; Paras, Katrina et al. (2018) ERBB3 and NGFR mark a distinct skeletal muscle progenitor cell in human development and hPSCs. Nat Cell Biol 20:46-57
Ferguson, Gabriel B; Van Handel, Ben; Bay, Maxwell et al. (2018) Mapping molecular landmarks of human skeletal ontogeny and pluripotent stem cell-derived articular chondrocytes. Nat Commun 9:3634
Young, Courtney S; Mokhonova, Ekaterina; Quinonez, Marbella et al. (2017) Creation of a Novel Humanized Dystrophic Mouse Model of Duchenne Muscular Dystrophy and Application of a CRISPR/Cas9 Gene Editing Therapy. J Neuromuscul Dis 4:139-145
Hazim, Roni A; Karumbayaram, Saravanan; Jiang, Mei et al. (2017) Differentiation of RPE cells from integration-free iPS cells and their cell biological characterization. Stem Cell Res Ther 8:217
Xi, Haibin; Fujiwara, Wakana; Gonzalez, Karen et al. (2017) In Vivo Human Somitogenesis Guides Somite Development from hPSCs. Cell Rep 18:1573-1585
Lee, Patrick C; Truong, Brian; Vega-Crespo, Agustin et al. (2016) Restoring Ureagenesis in Hepatocytes by CRISPR/Cas9-mediated Genomic Addition to Arginase-deficient Induced Pluripotent Stem Cells. Mol Ther Nucleic Acids 5:e394
Young, Courtney S; Hicks, Michael R; Ermolova, Natalia V et al. (2016) A Single CRISPR-Cas9 Deletion Strategy that Targets the Majority of DMD Patients Restores Dystrophin Function in hiPSC-Derived Muscle Cells. Cell Stem Cell 18:533-40
Hou, Shuang; Choi, Jin-sil; Chen, Kuan-Ju et al. (2015) Supramolecular nanosubstrate-mediated delivery for reprogramming and transdifferentiation of mammalian cells. Small 11:2499-504
Hicks, Michael; Pyle, April (2015) The Path from Pluripotency to Skeletal Muscle: Developmental Myogenesis Guides the Way. Cell Stem Cell 17:255-7
Peng, Jinliang; Garcia, Mitch André; Choi, Jin-sil et al. (2014) Molecular recognition enables nanosubstrate-mediated delivery of gene-encapsulated nanoparticles with high efficiency. ACS Nano 8:4621-9