Human pluripotent stem cells (hPSCs) have enormous promise for regenerative medicine as they can differentiate into cells from all three embryonic germ layers. Further hPSCs provide a unique pre-clinical screening tool for evaluating disease mechanisms and therapies in patient samples. Despite the enormous potential of hPSC, many obstacles need to be overcome before the use of stem cells in cell-based therapy will be realized, including an incomplete knowledge of the growth factor cues regulating human skeletal muscle progenitor cell (SMPC) specification, growth and engraftment. In this proposal we will investigate the cellular cues required to direct skeletal muscle fate from hPSCs as seen during embryonic development. We will also develop the first screening approach to identify new regulators of human SMPC fate. It is imperative that we understand how to obtain SMPCs without genetic or viral manipulation. As an alternative approach we have developed a unique non-viral reprogramming platform, which results in superior delivery and reprogramming efficiency, and will be utilized to specify SMPCs without viral or genetic manipulation. We will compare the in vivo engraftment potential of growth factor directed versus reprogrammed SMPCs in a mouse model of DMD. The development of reprogramming strategies to direct SMPC fate and the identification of signals regulating muscle progenitor cell specification and maintenance could improve our basic understanding of human skeletal myogenesis as well as enhance our ability to generate scalable progenitors from humans for muscle disorders including DMD.
Duchenne Muscular Dystrophy is a lethal muscle disease that results in progressive muscle loss. Human pluripotent stem cells (hPSCs) are unique in that they have the ability to differentiate any cell type in the human body including skeletal muscle progenitor cells. This grant will examine the role of intrinsic and extrinsic signals in directing muscle differentiation from hPSCs as well as development of a novel reprogramming platform for muscle differentiation, both of which have the potential to lead to development of human muscle for pre-clinical testing, regenerative applications, as well as improve our basic understanding of human muscle specification.
|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|