SIX1 reprograms myogenic transcription factors to maintain the rhabdomyosarcoma undifferentiated state

Jessica Y. Hsu; Etienne P. Danis; Stephanie Nance; Jenean H. O'Brien; Annika L. Gustafson; Veronica M. Wessells; Andrew E. Goodspeed; Jared C. Talbot; Sharon L. Amacher; Paul Jedlicka; Joshua C. Black; James C. Costello; Adam D. Durbin; Kristin B. Artinger; Heide L. Ford

doi:10.1016/j.celrep.2022.110323

SIX1 reprograms myogenic transcription factors to maintain the rhabdomyosarcoma undifferentiated state

Jessica Y. Hsu, Etienne P. Danis, Stephanie Nance, Jenean H. O'Brien, Annika L. Gustafson, Veronica M. Wessells, Andrew E. Goodspeed, Jared C. Talbot, Sharon L. Amacher, Paul Jedlicka, Joshua C. Black, James C. Costello, Adam D. Durbin, Kristin B. Artinger, Heide L. Ford

Research output: Contribution to journal › Article › peer-review

16 Scopus citations

Abstract

Rhabdomyosarcoma (RMS) is a pediatric muscle sarcoma characterized by expression of the myogenic lineage transcription factors (TFs) MYOD1 and MYOG. Despite high expression of these TFs, RMS cells fail to terminally differentiate, suggesting the presence of factors that alter their functions. Here, we demonstrate that the developmental TF SIX1 is highly expressed in RMS and critical for maintaining a muscle progenitor-like state. SIX1 loss induces differentiation of RMS cells into myotube-like cells and impedes tumor growth in vivo. We show that SIX1 maintains the RMS undifferentiated state by controlling enhancer activity and MYOD1 occupancy at loci more permissive to tumor growth over muscle differentiation. Finally, we demonstrate that a gene signature derived from SIX1 loss correlates with differentiation status and predicts RMS progression in human disease. Our findings demonstrate a master regulatory role of SIX1 in repression of RMS differentiation via genome-wide alterations in MYOD1 and MYOG-mediated transcription.

Original language	English (US)
Article number	110323
Journal	Cell reports
Volume	38
Issue number	5
DOIs	https://doi.org/10.1016/j.celrep.2022.110323
State	Published - Feb 1 2022
Externally published	Yes

Bibliographical note

Funding Information:
This work was supported by R21CA201809 (to H.L.F. and K.B.A.), R01CA224867 (to H.L.F.), R01CA183874 (to P.J.), GM117964 (to S.L.A.), NS098780 (to S.L.A.), K08CA245251 (to A.D.D.), an Alex’s Lemonade Stand Foundation Innovation Award (to H.L.F.), CU Cancer Center Molecular and Cellular Oncology Pilot Grant P30CA046934 (to H.L.F., K.B.A., and P.J.), and training fellowships T32GM763538 and TL1TR001081 (to J.Y.H.). The reported research was supported by NIH /NCI P30CA021765 (to St. Jude Children's Research Hospital Comprehensive Cancer Center), CureSearch for the Children’s Cancer Foundation, the Rally Foundation for Childhood Cancer Research, the American Lebanese Syrian Associated Charities (to A.D.D. and S.N.), a Pelotonia fellowship (to J.C.T.), and a University of Maine start-up fund (to J.C.T.). This work used the Cell Technologies, Functional Genomics, Pathology, and Biostatistics and Bioinformatics Shared Resource supported by P30CA046934.

Funding Information:
This work was supported by R21CA201809 (to H.L.F. and K.B.A.), R01CA224867 (to H.L.F.), R01CA183874 (to P.J.), GM117964 (to S.L.A.), NS098780 (to S.L.A.), K08CA245251 (to A.D.D.), an Alex's Lemonade Stand Foundation Innovation Award (to H.L.F.), CU Cancer Center Molecular and Cellular Oncology Pilot Grant P30CA046934 (to H.L.F. K.B.A. and P.J.), and training fellowships T32GM763538 and TL1TR001081 (to J.Y.H.). The reported research was supported by NIH/NCI P30CA021765 (to St. Jude Children's Research Hospital Comprehensive Cancer Center), CureSearch for the Children's Cancer Foundation, the Rally Foundation for Childhood Cancer Research, the American Lebanese Syrian Associated Charities (to A.D.D. and S.N.), a Pelotonia fellowship (to J.C.T.), and a University of Maine start-up fund (to J.C.T.). This work used the Cell Technologies, Functional Genomics, Pathology, and Biostatistics and Bioinformatics Shared Resource supported by P30CA046934. K.B.A. and H.L.F. conceptualized and supervised experiments with input from A.D.D. J.C.B. and J.C.C. J.Y.H. S.N. and A.L.G. performed cell line experiments and were involved in data generation and interpretation. J.Y.H. performed mouse experiments. J.Y.H. and J.H.O. performed zebrafish experiments. J.Y.H. S.N. E.P.D. and A.E.G. performed and analyzed NGS experiments. V.M.W. and P.J. performed and scored tumor histology in zebrafish and mouse models. Critical reagents were provided by S.L.A. A.D.D. and J.T. J.Y.H. wrote the manuscript with significant input from K.B.A. H.L.F. A.D.D. J.C.B. and J.C.T. All authors contributed to manuscript editing and review. J.C.C. is a co-founder of PrecisionProfile. H.L.F. is a co-founder of Sieyax, LLC.

Publisher Copyright:
© 2022 The Authors

Keywords

chromatin
CUT&RUN
mouse xenograft
muscle differentiation
muscle progenitor
MYOD1
rhabdomyosarcoma
SIX1
transcriptional control
zebrafish

PubMed: MeSH publication types

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.celrep.2022.110323

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Hsu, J. Y., Danis, E. P., Nance, S., O'Brien, J. H., Gustafson, A. L., Wessells, V. M., Goodspeed, A. E., Talbot, J. C., Amacher, S. L., Jedlicka, P., Black, J. C., Costello, J. C., Durbin, A. D., Artinger, K. B., & Ford, H. L. (2022). SIX1 reprograms myogenic transcription factors to maintain the rhabdomyosarcoma undifferentiated state. Cell reports, 38(5), Article 110323. https://doi.org/10.1016/j.celrep.2022.110323

Hsu, JY, Danis, EP, Nance, S, O'Brien, JH, Gustafson, AL, Wessells, VM, Goodspeed, AE, Talbot, JC, Amacher, SL, Jedlicka, P, Black, JC, Costello, JC, Durbin, AD, Artinger, KB & Ford, HL 2022, 'SIX1 reprograms myogenic transcription factors to maintain the rhabdomyosarcoma undifferentiated state', Cell reports, vol. 38, no. 5, 110323. https://doi.org/10.1016/j.celrep.2022.110323

@article{aa5f03266e76408f918c047bd40b5a3e,

title = "SIX1 reprograms myogenic transcription factors to maintain the rhabdomyosarcoma undifferentiated state",

abstract = "Rhabdomyosarcoma (RMS) is a pediatric muscle sarcoma characterized by expression of the myogenic lineage transcription factors (TFs) MYOD1 and MYOG. Despite high expression of these TFs, RMS cells fail to terminally differentiate, suggesting the presence of factors that alter their functions. Here, we demonstrate that the developmental TF SIX1 is highly expressed in RMS and critical for maintaining a muscle progenitor-like state. SIX1 loss induces differentiation of RMS cells into myotube-like cells and impedes tumor growth in vivo. We show that SIX1 maintains the RMS undifferentiated state by controlling enhancer activity and MYOD1 occupancy at loci more permissive to tumor growth over muscle differentiation. Finally, we demonstrate that a gene signature derived from SIX1 loss correlates with differentiation status and predicts RMS progression in human disease. Our findings demonstrate a master regulatory role of SIX1 in repression of RMS differentiation via genome-wide alterations in MYOD1 and MYOG-mediated transcription.",

keywords = "chromatin, CUT&RUN, mouse xenograft, muscle differentiation, muscle progenitor, MYOD1, rhabdomyosarcoma, SIX1, transcriptional control, zebrafish",

author = "Hsu, {Jessica Y.} and Danis, {Etienne P.} and Stephanie Nance and O'Brien, {Jenean H.} and Gustafson, {Annika L.} and Wessells, {Veronica M.} and Goodspeed, {Andrew E.} and Talbot, {Jared C.} and Amacher, {Sharon L.} and Paul Jedlicka and Black, {Joshua C.} and Costello, {James C.} and Durbin, {Adam D.} and Artinger, {Kristin B.} and Ford, {Heide L.}",

note = "Funding Information: This work was supported by R21CA201809 (to H.L.F. and K.B.A.), R01CA224867 (to H.L.F.), R01CA183874 (to P.J.), GM117964 (to S.L.A.), NS098780 (to S.L.A.), K08CA245251 (to A.D.D.), an Alex{\textquoteright}s Lemonade Stand Foundation Innovation Award (to H.L.F.), CU Cancer Center Molecular and Cellular Oncology Pilot Grant P30CA046934 (to H.L.F., K.B.A., and P.J.), and training fellowships T32GM763538 and TL1TR001081 (to J.Y.H.). The reported research was supported by NIH /NCI P30CA021765 (to St. Jude Children's Research Hospital Comprehensive Cancer Center), CureSearch for the Children{\textquoteright}s Cancer Foundation, the Rally Foundation for Childhood Cancer Research, the American Lebanese Syrian Associated Charities (to A.D.D. and S.N.), a Pelotonia fellowship (to J.C.T.), and a University of Maine start-up fund (to J.C.T.). This work used the Cell Technologies, Functional Genomics, Pathology, and Biostatistics and Bioinformatics Shared Resource supported by P30CA046934. Funding Information: This work was supported by R21CA201809 (to H.L.F. and K.B.A.), R01CA224867 (to H.L.F.), R01CA183874 (to P.J.), GM117964 (to S.L.A.), NS098780 (to S.L.A.), K08CA245251 (to A.D.D.), an Alex's Lemonade Stand Foundation Innovation Award (to H.L.F.), CU Cancer Center Molecular and Cellular Oncology Pilot Grant P30CA046934 (to H.L.F. K.B.A. and P.J.), and training fellowships T32GM763538 and TL1TR001081 (to J.Y.H.). The reported research was supported by NIH/NCI P30CA021765 (to St. Jude Children's Research Hospital Comprehensive Cancer Center), CureSearch for the Children's Cancer Foundation, the Rally Foundation for Childhood Cancer Research, the American Lebanese Syrian Associated Charities (to A.D.D. and S.N.), a Pelotonia fellowship (to J.C.T.), and a University of Maine start-up fund (to J.C.T.). This work used the Cell Technologies, Functional Genomics, Pathology, and Biostatistics and Bioinformatics Shared Resource supported by P30CA046934. K.B.A. and H.L.F. conceptualized and supervised experiments with input from A.D.D. J.C.B. and J.C.C. J.Y.H. S.N. and A.L.G. performed cell line experiments and were involved in data generation and interpretation. J.Y.H. performed mouse experiments. J.Y.H. and J.H.O. performed zebrafish experiments. J.Y.H. S.N. E.P.D. and A.E.G. performed and analyzed NGS experiments. V.M.W. and P.J. performed and scored tumor histology in zebrafish and mouse models. Critical reagents were provided by S.L.A. A.D.D. and J.T. J.Y.H. wrote the manuscript with significant input from K.B.A. H.L.F. A.D.D. J.C.B. and J.C.T. All authors contributed to manuscript editing and review. J.C.C. is a co-founder of PrecisionProfile. H.L.F. is a co-founder of Sieyax, LLC. Publisher Copyright: {\textcopyright} 2022 The Authors",

year = "2022",

month = feb,

day = "1",

doi = "10.1016/j.celrep.2022.110323",

language = "English (US)",

volume = "38",

journal = "Cell reports",

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TY - JOUR

T1 - SIX1 reprograms myogenic transcription factors to maintain the rhabdomyosarcoma undifferentiated state

AU - Hsu, Jessica Y.

AU - Danis, Etienne P.

AU - Nance, Stephanie

AU - O'Brien, Jenean H.

AU - Gustafson, Annika L.

AU - Wessells, Veronica M.

AU - Goodspeed, Andrew E.

AU - Talbot, Jared C.

AU - Amacher, Sharon L.

AU - Jedlicka, Paul

AU - Black, Joshua C.

AU - Costello, James C.

AU - Durbin, Adam D.

AU - Artinger, Kristin B.

AU - Ford, Heide L.

N1 - Funding Information: This work was supported by R21CA201809 (to H.L.F. and K.B.A.), R01CA224867 (to H.L.F.), R01CA183874 (to P.J.), GM117964 (to S.L.A.), NS098780 (to S.L.A.), K08CA245251 (to A.D.D.), an Alex’s Lemonade Stand Foundation Innovation Award (to H.L.F.), CU Cancer Center Molecular and Cellular Oncology Pilot Grant P30CA046934 (to H.L.F., K.B.A., and P.J.), and training fellowships T32GM763538 and TL1TR001081 (to J.Y.H.). The reported research was supported by NIH /NCI P30CA021765 (to St. Jude Children's Research Hospital Comprehensive Cancer Center), CureSearch for the Children’s Cancer Foundation, the Rally Foundation for Childhood Cancer Research, the American Lebanese Syrian Associated Charities (to A.D.D. and S.N.), a Pelotonia fellowship (to J.C.T.), and a University of Maine start-up fund (to J.C.T.). This work used the Cell Technologies, Functional Genomics, Pathology, and Biostatistics and Bioinformatics Shared Resource supported by P30CA046934. Funding Information: This work was supported by R21CA201809 (to H.L.F. and K.B.A.), R01CA224867 (to H.L.F.), R01CA183874 (to P.J.), GM117964 (to S.L.A.), NS098780 (to S.L.A.), K08CA245251 (to A.D.D.), an Alex's Lemonade Stand Foundation Innovation Award (to H.L.F.), CU Cancer Center Molecular and Cellular Oncology Pilot Grant P30CA046934 (to H.L.F. K.B.A. and P.J.), and training fellowships T32GM763538 and TL1TR001081 (to J.Y.H.). The reported research was supported by NIH/NCI P30CA021765 (to St. Jude Children's Research Hospital Comprehensive Cancer Center), CureSearch for the Children's Cancer Foundation, the Rally Foundation for Childhood Cancer Research, the American Lebanese Syrian Associated Charities (to A.D.D. and S.N.), a Pelotonia fellowship (to J.C.T.), and a University of Maine start-up fund (to J.C.T.). This work used the Cell Technologies, Functional Genomics, Pathology, and Biostatistics and Bioinformatics Shared Resource supported by P30CA046934. K.B.A. and H.L.F. conceptualized and supervised experiments with input from A.D.D. J.C.B. and J.C.C. J.Y.H. S.N. and A.L.G. performed cell line experiments and were involved in data generation and interpretation. J.Y.H. performed mouse experiments. J.Y.H. and J.H.O. performed zebrafish experiments. J.Y.H. S.N. E.P.D. and A.E.G. performed and analyzed NGS experiments. V.M.W. and P.J. performed and scored tumor histology in zebrafish and mouse models. Critical reagents were provided by S.L.A. A.D.D. and J.T. J.Y.H. wrote the manuscript with significant input from K.B.A. H.L.F. A.D.D. J.C.B. and J.C.T. All authors contributed to manuscript editing and review. J.C.C. is a co-founder of PrecisionProfile. H.L.F. is a co-founder of Sieyax, LLC. Publisher Copyright: © 2022 The Authors

PY - 2022/2/1

Y1 - 2022/2/1

N2 - Rhabdomyosarcoma (RMS) is a pediatric muscle sarcoma characterized by expression of the myogenic lineage transcription factors (TFs) MYOD1 and MYOG. Despite high expression of these TFs, RMS cells fail to terminally differentiate, suggesting the presence of factors that alter their functions. Here, we demonstrate that the developmental TF SIX1 is highly expressed in RMS and critical for maintaining a muscle progenitor-like state. SIX1 loss induces differentiation of RMS cells into myotube-like cells and impedes tumor growth in vivo. We show that SIX1 maintains the RMS undifferentiated state by controlling enhancer activity and MYOD1 occupancy at loci more permissive to tumor growth over muscle differentiation. Finally, we demonstrate that a gene signature derived from SIX1 loss correlates with differentiation status and predicts RMS progression in human disease. Our findings demonstrate a master regulatory role of SIX1 in repression of RMS differentiation via genome-wide alterations in MYOD1 and MYOG-mediated transcription.

AB - Rhabdomyosarcoma (RMS) is a pediatric muscle sarcoma characterized by expression of the myogenic lineage transcription factors (TFs) MYOD1 and MYOG. Despite high expression of these TFs, RMS cells fail to terminally differentiate, suggesting the presence of factors that alter their functions. Here, we demonstrate that the developmental TF SIX1 is highly expressed in RMS and critical for maintaining a muscle progenitor-like state. SIX1 loss induces differentiation of RMS cells into myotube-like cells and impedes tumor growth in vivo. We show that SIX1 maintains the RMS undifferentiated state by controlling enhancer activity and MYOD1 occupancy at loci more permissive to tumor growth over muscle differentiation. Finally, we demonstrate that a gene signature derived from SIX1 loss correlates with differentiation status and predicts RMS progression in human disease. Our findings demonstrate a master regulatory role of SIX1 in repression of RMS differentiation via genome-wide alterations in MYOD1 and MYOG-mediated transcription.

KW - chromatin

KW - CUT&RUN

KW - mouse xenograft

KW - muscle differentiation

KW - muscle progenitor

KW - MYOD1

KW - rhabdomyosarcoma

KW - SIX1

KW - transcriptional control

KW - zebrafish

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U2 - 10.1016/j.celrep.2022.110323

DO - 10.1016/j.celrep.2022.110323

M3 - Article

C2 - 35108532

AN - SCOPUS:85123783812

SN - 2211-1247

VL - 38

JO - Cell reports

JF - Cell reports

IS - 5

M1 - 110323

ER -

SIX1 reprograms myogenic transcription factors to maintain the rhabdomyosarcoma undifferentiated state

Abstract

Bibliographical note

Keywords

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UN SDGs

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