Adult muscle stem cells (MuSC), also known as satellite cells, are the main source skeletal muscle regeneration. This is due to their ability to self-renew in vivo, i.e. to generate upon cell division more copies of themselves as well as give rise to committed progeny, as we have conclusively demonstrated by single MuSC transplantation. MuSC exist in healthy adult tissues in a quiescent state, and upon stress or injury they are activated to proliferate and generate large numbers of progenitors to repair the damaged tissue. Although the balance between quiescence and activation is a critical switch in MuSC function, the molecular mechanisms regulating this transition are still largely unknown. A more clear understanding of the regulatory networks underlying MuSC activation might provide novel targets that would aid in the development of therapeutic approaches to ameliorate muscle-wasting diseases. Our preliminary findings indicate for the first time that STAT3 plays a direct role in the transcriptional activation of the bHLH myogenic regulatory factor MyoD, a key event as MuSC exit the quiescent state. STAT3 is a transcription factor that plays a major role in self-renewal of several types of stem cells, including embryonic, intestinal and hematopoietic compartments. Upon cytokine stimulation, STAT3 is phosphorylated by JAK kinases, it homodimerizes, translocates to the nucleus and binds DNA to activate the transcription of target genes. Although it has been previously shown that STAT3 is activated upon skeletal muscle injury, its role in MuSC activation as well as its relevant downstream targets are still poorly understood. Accordingly, the focus of this proposal is to investigate the role of the JAK/STAT3 pathway in MuSC activation and its functional interaction with MyoD. Our research will take advantage of the following tools: (1) Loss of function studies in conjunction with time-lapse microscopy, to monitor MuSC activation, proliferation and survival in vitro, (2) Luciferase reporter assay using deletion mutants of the MyoD proximal enhancer, to validate the direct role of STAT3 in MyoD transcriptional activation, (3) Conditional genetic ablation of STAT3 in MuSC, in order to evaluate its role in MuSC function in vivo in the intact animal. Together, these studie would constitute a conceptual advancement in the field, as they would identify a direct functional interaction between JAK/STAT3 and MyoD, and further extend our knowledge of the regulatory network in MuSC activation. Finally, these findings would aid in the development of strategies aimed at promoting muscle stem cell-mediated tissue regeneration to ameliorate muscle-wasting diseases.
The proposed studies are relevant to efforts for developing therapeutic approaches for muscle-wasting diseases. We will dissect molecular mechanisms regulating skeletal muscle stem cell activation, thus providing a foundation of knowledge on how muscle stem cells coordinate tissue regeneration and how they can rapidly respond to stress or injury and repair the damaged tissue. Finally, these studies will suggest novel targets that will aid in the development of novel therapies to ameliorate muscle-wasting diseases.
|Tierney, Matthew T; Sacco, Alessandra (2016) Inducing and Evaluating Skeletal Muscle Injury by Notexin and Barium Chloride. Methods Mol Biol 1460:53-60|
|Sala, David; Sacco, Alessandra (2016) Signal transducer and activator of transcription 3 signaling as a potential target to treat muscle wasting diseases. Curr Opin Clin Nutr Metab Care 19:171-6|
|Tierney, Matthew T; Sacco, Alessandra (2016) Satellite Cell Heterogeneity in Skeletal Muscle Homeostasis. Trends Cell Biol 26:434-444|
|Sacco, Alessandra; Puri, Pier Lorenzo (2015) Regulation of Muscle Satellite Cell Function in Tissue Homeostasis and Aging. Cell Stem Cell 16:585-7|
|Gromova, Anastasia; Tierney, Matthew T; Sacco, Alessandra (2015) FACS-based Satellite Cell Isolation From Mouse Hind Limb Muscles. Bio Protoc 5:|
|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|