Adult stem cells are capable of self renewal and differentiation. Although they possess self-renewal properties, this potential is not limitless. This begs the following question: how are stem cells maintained throughout life? The answer lies in the reversible state of quiescence. Adult stem cells are predominantly in a quiescent state interspersed with rare traverses into the cell cycle unlike their downstream descendents that spend more time in cycle. Using cell labeling techniques to monitor cell division history, stem cells that divide less frequently are enriched for self-renewal potential. In muscular dystrophies, muscle stem cells (satellite cells, SCs) undergo continuous bouts of proliferation, which leads to a depletion of the stem cell pool over time. In aged muscle the SC pool is diminished and displays impaired renewal and differentiation. Therefore understanding how 'proliferation-restricted'stem cells may hold the key to optimizing stem cell function during aging and disease. In this grant proposal we will interrogate 1) the functional heterogeneity between slow and faster dividing SCs. 2) How the slow dividing SC population is specified and maintained throughout life through Spry, an inhibitor of growth factor signaling. 3) The consequences of losing slow dividing subset of SCs SC function and muscle homeostasis during aging. Slow dividing SCs will be identified in vivo based on label retaining character (LRC) using a transgenic H2B-GFP approach;cells that undergo fewer divisions retain more label. Satellite cells will be characterized for LRC and non-LRC;both subpopulations will be tested for their functional differences based on in vivo and in vitro assays. We have generated preliminary data that demonstrates Spry1 expression is enriched in LRCs and genetic disruption of Spry1 leads to a loss of LRC in growing muscle and an accelerated decline in the number of SCs during aging. We will extend these studies to delete and overexpress Spry1 specifically in SCs in a temporally inducible manner. We will disrupt Spry1 to alter LRC establishment and maintenance and study the effects on SC function and muscle phenotype.
The specific aims of this proposal are: 1) To study the slow division dynamics of satellite cells during developmental and postnatal myogenesis, 2) to identify whether Spry1 is required and sufficient to regulate LRC SCs, and 3) to understand the consequences of losing SC LRC throughout life.

Public Health Relevance

Identifying a specialized subset of highly functioning muscle stem cells is essential to ameliorate skeletal muscle aging and diseases. Unfortunately, the identity of such cells and how they are established and maintained in not known. In this project we will identify highly functioning SCs and dissect an essential genetic coordinator that defines their functional superiority. We will employ genetic strategies to impair or enhance this SC subset early in life and study the long term effects in aged muscle. Deciphering endogenous stem cell modifiers that can be modulated to maintain stem cell function and tissue homeostasis throughout life is critical for harnessing their potential to treat muscle related diseases. Furthermore, the biology of satellite cells may serve as a paradigm for stem cells in other tissues.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR061002-02
Application #
8442867
Study Section
Development - 2 Study Section (DEV2)
Program Officer
Boyce, Amanda T
Project Start
2012-04-01
Project End
2017-03-31
Budget Start
2013-04-01
Budget End
2014-03-31
Support Year
2
Fiscal Year
2013
Total Cost
$333,963
Indirect Cost
$120,213
Name
Massachusetts General Hospital
Department
Type
DUNS #
073130411
City
Boston
State
MA
Country
United States
Zip Code
02199
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Brack, Andrew S; Muñoz-Cánoves, Pura (2016) The ins and outs of muscle stem cell aging. Skelet Muscle 6:1
Egerman, Marc A; Cadena, Samuel M; Gilbert, Jason A et al. (2015) GDF11 Increases with Age and Inhibits Skeletal Muscle Regeneration. Cell Metab 22:164-74
Kollu, Swapna; Abou-Khalil, Rana; Shen, Carl et al. (2015) The Spindle Assembly Checkpoint Safeguards Genomic Integrity of Skeletal Muscle Satellite Cells. Stem Cell Reports 4:1061-74
Jung, Yunjoon; Brack, Andrew S (2014) Cellular mechanisms of somatic stem cell aging. Curr Top Dev Biol 107:405-38
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
Abraham, Jinu; Nuñez-Álvarez, Yaiza; Hettmer, Simone et al. (2014) Lineage of origin in rhabdomyosarcoma informs pharmacological response. Genes Dev 28:1578-91
Brack, Andrew S (2014) Pax7 is back. Skelet Muscle 4:24
Brack, Andrew S; Hochedlinger, Konrad (2013) ISSCR 2013: back to Bean Town. Stem Cell Reports 1:479-85
Chakkalakal, Joe V; Jones, Kieran M; Basson, M Albert et al. (2012) The aged niche disrupts muscle stem cell quiescence. Nature 490:355-60

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