Muscle stem cells (MuSC) are required for the development and repair of skeletal muscle. During these dynamic periods of muscle growth and regeneration, MuSC must selectively interpret a host of signaling molecules to self-renew. This ability is progressively impaired in Duchenne muscular dystrophy (DMD), a devastating muscle wasting disease for which there is no cure. DMD is characterized by altered microenvironmental signaling that negatively influences MuSC-mediated regenerative potential. To better understand how MuSC communicate with their extracellular environment, I explored several membrane-bound mediators of signal transduction. Heparan sulfate proteoglycans (HSPG), in particular syndecans, are highly expressed in MuSC and can regulate ligand bioavailability and receptor formation. Heterogeneous sulfation patterns on HSPG via post-translational modifications increase behavioral complexity, leading to a differential activity of critical signalng pathways. Because regeneration in the adult recapitulates several aspects of myogenesis, I focused my preliminary studies on HSPG sulfation in fetal MuSC, the developmental precursors of adult MuSC. I have demonstrated that Sulf1, a specific regulator of HSPG 6-O-sulfation, is downregulated in fetal MuSC. I provide evidence that fetal MuSC are resistant to myogenic commitment in vitro and capable of rapid in vivo expansion and long-term self-renewal following transplantation. Furthermore, uncommitted fetal MuSC are preferentially sensitive to the potent mitogen FGF2 while less responsive to Wnt/?-catenin-mediated myogenic differentiation. Indeed, Sulf1 has been previously shown to simultaneously promote Wnt/?-catenin and inhibit FGF2 signaling. Sulf1 and HSPG expression are dysregulated in muscle wasting conditions. Therefore, the goal of this project is to examine the role of HSPG 6-O-sulfation on the regulation of dystrophic MuSC self-renewal and regenerative potential. To accomplish this, I propose (Aim 1a) to validate HSPG sulfatase and FGF and Wnt/?-catenin signaling component expression levels throughout myogenesis and in dystrophic animals. To determine MuSC responsiveness to FGF and Wnt/?-catenin pathway activation (Aim 1b), I will measure the expression and phosphorylation levels of downstream signaling pathway effectors. Inhibition of HSPG 6-O-sulfation will ascertain its role in FGF receptor complex formation and Wnt3a ligand bioavailability, defining the mechanisms underlying HSPG regulation of proper pathway activation.
(Aim 1 c). I will determine the impact of Sulf1 inhibition on dystrophic, myofiber-associated MuSC self-renewal (Aim 2a). Finally, I will evaluate the functional role of HSPG 6-O-sulfation in a preclinical model of DMD by inhibiting Sulf1 expression in (Aim 2b) transplanted, donor MuSC and (Aim 2c) native, dystrophic MuSC. Through these studies I will investigate a molecular mechanism, HSPG 6-O-sulfation, able to regulate FGF and Wnt/?-catenin signaling and MuSC regenerative potential in dystrophic animals. This work will potentially identify Sulf1 as a novel therapeutic target for the enhancement of MuSC self-renewal and the amelioration of DMD.
Muscle stem cells progressively lose their regenerative potential in muscle wasting conditions. This is caused in part by the communication of unfavorable signaling cues resulting from a diseased microenvironment. The proposed studies will (1) provide insight into the molecular mechanisms underlying the means by which muscle stem cells interpret external cues to regulate their behavior and (2) aid in the development of interventions designed to ameliorate muscle stem cell impairment in Duchenne muscular dystrophy.
|Tierney, Matthew Timothy; Gromova, Anastasia; Sesillo, Francesca Boscolo et al. (2016) Autonomous Extracellular Matrix Remodeling Controls a Progressive Adaptation in Muscle Stem Cell Regenerative Capacity during Development. Cell Rep 14:1940-52|
|Tierney, Matthew T; Sacco, Alessandra (2016) Satellite Cell Heterogeneity in Skeletal Muscle Homeostasis. Trends Cell Biol 26:434-44|
|Gromova, Anastasia; Tierney, Matthew T; Sacco, Alessandra (2015) FACS-based Satellite Cell Isolation From Mouse Hind Limb Muscles. Bio Protoc 5:|
|Kang, Yunyi; Tierney, Matthew; Ong, Edison et al. (2015) Combinations of Kinase Inhibitors Protecting Myoblasts against Hypoxia. PLoS One 10:e0126718|
|Tierney, Matthew Timothy; Aydogdu, Tufan; Sala, David et al. (2014) STAT3 signaling controls satellite cell expansion and skeletal muscle repair. Nat Med 20:1182-6|
|Chakkalakal, Joe V; Christensen, Josef; Xiang, Wanyi et al. (2014) Early forming label-retaining muscle stem cells require p27kip1 for maintenance of the primitive state. Development 141:1649-59|
|Gokhin, David S; Tierney, Matthew T; Sui, Zhenhua et al. (2014) Calpain-mediated proteolysis of tropomodulin isoforms leads to thin filament elongation in dystrophic skeletal muscle. Mol Biol Cell 25:852-65|