Preclinical rationale for entinostat in embryonal rhabdomyosarcoma

Narendra Bharathy, Noah E. Berlow, Eric Wang, Jinu Abraham, Teagan P. Settelmeyer, Jody Hooper, Matthew N. Svalina, Zia Bajwa, Martin W. Goros, Brian S. Hernandez, Johannes E. Wolff, Ranadip Pal, Angela M. Davies, Arya Ashok, Darnell Bushby, Maria Mancini, Christopher Noakes, Neal C. Goodwin, Peter Ordentlich, James KeckDouglas S. Hawkins, Erin R. Rudzinski, Atiya Mansoor, Theodore J. Perkins, Christopher R. Vakoc, Joel E. Michalek, Charles Keller

Research output: Contribution to journalArticle

Abstract

Background: Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in the pediatric cancer population. Survival among metastatic RMS patients has remained dismal yet unimproved for years. We previously identified the class I-specific histone deacetylase inhibitor, entinostat (ENT), as a pharmacological agent that transcriptionally suppresses the PAX3:FOXO1 tumor-initiating fusion gene found in alveolar rhabdomyosarcoma (aRMS), and we further investigated the mechanism by which ENT suppresses PAX3:FOXO1 oncogene and demonstrated the preclinical efficacy of ENT in RMS orthotopic allograft and patient-derived xenograft (PDX) models. In this study, we investigated whether ENT also has antitumor activity in fusion-negative eRMS orthotopic allografts and PDX models either as a single agent or in combination with vincristine (VCR). Methods: We tested the efficacy of ENT and VCR as single agents and in combination in orthotopic allograft and PDX mouse models of eRMS. We then performed CRISPR screening to identify which HDAC among the class I HDACs is responsible for tumor growth inhibition in eRMS. To analyze whether ENT treatment as a single agent or in combination with VCR induces myogenic differentiation, we performed hematoxylin and eosin (H&E) staining in tumors. Results: ENT in combination with the chemotherapy VCR has synergistic antitumor activity in a subset of fusion-negative eRMS in orthotopic "allografts," although PDX mouse models were too hypersensitive to the VCR dose used to detect synergy. Mechanistic studies involving CRISPR suggest that HDAC3 inhibition is the primary mechanism of cell-autonomous cytoreduction in eRMS. Following cytoreduction in vivo, residual tumor cells in the allograft models treated with chemotherapy undergo a dramatic, entinostat-induced (70-100%) conversion to non-proliferative rhabdomyoblasts. Conclusion: Our results suggest that the targeting class I HDACs may provide a therapeutic benefit for selected patients with eRMS. ENT's preclinical in vivo efficacy makes ENT a rational drug candidate in a phase II clinical trial for eRMS.

Original languageEnglish (US)
Article number12
JournalSkeletal Muscle
Volume9
Issue number1
DOIs
StatePublished - May 21 2019

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Embryonal Rhabdomyosarcoma
Vincristine
Allografts
Heterografts
Rhabdomyosarcoma
Clustered Regularly Interspaced Short Palindromic Repeats
Neoplasms
Alveolar Rhabdomyosarcoma
entinostat
Phase II Clinical Trials
Histone Deacetylase Inhibitors
Gene Fusion
Residual Neoplasm
Hematoxylin
Eosine Yellowish-(YS)
Combination Drug Therapy
Oncogenes
Sarcoma
Pharmacology
Pediatrics

Keywords

  • Embryonal rhabdomyosarcoma (eRMS)
  • Entinostat
  • HDAC3
  • Patient-derived xenograft (PDX)
  • Vincristine

ASJC Scopus subject areas

  • Orthopedics and Sports Medicine
  • Molecular Biology
  • Cell Biology

Cite this

Bharathy, N., Berlow, N. E., Wang, E., Abraham, J., Settelmeyer, T. P., Hooper, J., ... Keller, C. (2019). Preclinical rationale for entinostat in embryonal rhabdomyosarcoma. Skeletal Muscle, 9(1), [12]. https://doi.org/10.1186/s13395-019-0198-x

Preclinical rationale for entinostat in embryonal rhabdomyosarcoma. / Bharathy, Narendra; Berlow, Noah E.; Wang, Eric; Abraham, Jinu; Settelmeyer, Teagan P.; Hooper, Jody; Svalina, Matthew N.; Bajwa, Zia; Goros, Martin W.; Hernandez, Brian S.; Wolff, Johannes E.; Pal, Ranadip; Davies, Angela M.; Ashok, Arya; Bushby, Darnell; Mancini, Maria; Noakes, Christopher; Goodwin, Neal C.; Ordentlich, Peter; Keck, James; Hawkins, Douglas S.; Rudzinski, Erin R.; Mansoor, Atiya; Perkins, Theodore J.; Vakoc, Christopher R.; Michalek, Joel E.; Keller, Charles.

In: Skeletal Muscle, Vol. 9, No. 1, 12, 21.05.2019.

Research output: Contribution to journalArticle

Bharathy, N, Berlow, NE, Wang, E, Abraham, J, Settelmeyer, TP, Hooper, J, Svalina, MN, Bajwa, Z, Goros, MW, Hernandez, BS, Wolff, JE, Pal, R, Davies, AM, Ashok, A, Bushby, D, Mancini, M, Noakes, C, Goodwin, NC, Ordentlich, P, Keck, J, Hawkins, DS, Rudzinski, ER, Mansoor, A, Perkins, TJ, Vakoc, CR, Michalek, JE & Keller, C 2019, 'Preclinical rationale for entinostat in embryonal rhabdomyosarcoma', Skeletal Muscle, vol. 9, no. 1, 12. https://doi.org/10.1186/s13395-019-0198-x
Bharathy N, Berlow NE, Wang E, Abraham J, Settelmeyer TP, Hooper J et al. Preclinical rationale for entinostat in embryonal rhabdomyosarcoma. Skeletal Muscle. 2019 May 21;9(1). 12. https://doi.org/10.1186/s13395-019-0198-x
Bharathy, Narendra ; Berlow, Noah E. ; Wang, Eric ; Abraham, Jinu ; Settelmeyer, Teagan P. ; Hooper, Jody ; Svalina, Matthew N. ; Bajwa, Zia ; Goros, Martin W. ; Hernandez, Brian S. ; Wolff, Johannes E. ; Pal, Ranadip ; Davies, Angela M. ; Ashok, Arya ; Bushby, Darnell ; Mancini, Maria ; Noakes, Christopher ; Goodwin, Neal C. ; Ordentlich, Peter ; Keck, James ; Hawkins, Douglas S. ; Rudzinski, Erin R. ; Mansoor, Atiya ; Perkins, Theodore J. ; Vakoc, Christopher R. ; Michalek, Joel E. ; Keller, Charles. / Preclinical rationale for entinostat in embryonal rhabdomyosarcoma. In: Skeletal Muscle. 2019 ; Vol. 9, No. 1.
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abstract = "Background: Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in the pediatric cancer population. Survival among metastatic RMS patients has remained dismal yet unimproved for years. We previously identified the class I-specific histone deacetylase inhibitor, entinostat (ENT), as a pharmacological agent that transcriptionally suppresses the PAX3:FOXO1 tumor-initiating fusion gene found in alveolar rhabdomyosarcoma (aRMS), and we further investigated the mechanism by which ENT suppresses PAX3:FOXO1 oncogene and demonstrated the preclinical efficacy of ENT in RMS orthotopic allograft and patient-derived xenograft (PDX) models. In this study, we investigated whether ENT also has antitumor activity in fusion-negative eRMS orthotopic allografts and PDX models either as a single agent or in combination with vincristine (VCR). Methods: We tested the efficacy of ENT and VCR as single agents and in combination in orthotopic allograft and PDX mouse models of eRMS. We then performed CRISPR screening to identify which HDAC among the class I HDACs is responsible for tumor growth inhibition in eRMS. To analyze whether ENT treatment as a single agent or in combination with VCR induces myogenic differentiation, we performed hematoxylin and eosin (H&E) staining in tumors. Results: ENT in combination with the chemotherapy VCR has synergistic antitumor activity in a subset of fusion-negative eRMS in orthotopic {"}allografts,{"} although PDX mouse models were too hypersensitive to the VCR dose used to detect synergy. Mechanistic studies involving CRISPR suggest that HDAC3 inhibition is the primary mechanism of cell-autonomous cytoreduction in eRMS. Following cytoreduction in vivo, residual tumor cells in the allograft models treated with chemotherapy undergo a dramatic, entinostat-induced (70-100{\%}) conversion to non-proliferative rhabdomyoblasts. Conclusion: Our results suggest that the targeting class I HDACs may provide a therapeutic benefit for selected patients with eRMS. ENT's preclinical in vivo efficacy makes ENT a rational drug candidate in a phase II clinical trial for eRMS.",
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T1 - Preclinical rationale for entinostat in embryonal rhabdomyosarcoma

AU - Bharathy, Narendra

AU - Berlow, Noah E.

AU - Wang, Eric

AU - Abraham, Jinu

AU - Settelmeyer, Teagan P.

AU - Hooper, Jody

AU - Svalina, Matthew N.

AU - Bajwa, Zia

AU - Goros, Martin W.

AU - Hernandez, Brian S.

AU - Wolff, Johannes E.

AU - Pal, Ranadip

AU - Davies, Angela M.

AU - Ashok, Arya

AU - Bushby, Darnell

AU - Mancini, Maria

AU - Noakes, Christopher

AU - Goodwin, Neal C.

AU - Ordentlich, Peter

AU - Keck, James

AU - Hawkins, Douglas S.

AU - Rudzinski, Erin R.

AU - Mansoor, Atiya

AU - Perkins, Theodore J.

AU - Vakoc, Christopher R.

AU - Michalek, Joel E.

AU - Keller, Charles

PY - 2019/5/21

Y1 - 2019/5/21

N2 - Background: Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in the pediatric cancer population. Survival among metastatic RMS patients has remained dismal yet unimproved for years. We previously identified the class I-specific histone deacetylase inhibitor, entinostat (ENT), as a pharmacological agent that transcriptionally suppresses the PAX3:FOXO1 tumor-initiating fusion gene found in alveolar rhabdomyosarcoma (aRMS), and we further investigated the mechanism by which ENT suppresses PAX3:FOXO1 oncogene and demonstrated the preclinical efficacy of ENT in RMS orthotopic allograft and patient-derived xenograft (PDX) models. In this study, we investigated whether ENT also has antitumor activity in fusion-negative eRMS orthotopic allografts and PDX models either as a single agent or in combination with vincristine (VCR). Methods: We tested the efficacy of ENT and VCR as single agents and in combination in orthotopic allograft and PDX mouse models of eRMS. We then performed CRISPR screening to identify which HDAC among the class I HDACs is responsible for tumor growth inhibition in eRMS. To analyze whether ENT treatment as a single agent or in combination with VCR induces myogenic differentiation, we performed hematoxylin and eosin (H&E) staining in tumors. Results: ENT in combination with the chemotherapy VCR has synergistic antitumor activity in a subset of fusion-negative eRMS in orthotopic "allografts," although PDX mouse models were too hypersensitive to the VCR dose used to detect synergy. Mechanistic studies involving CRISPR suggest that HDAC3 inhibition is the primary mechanism of cell-autonomous cytoreduction in eRMS. Following cytoreduction in vivo, residual tumor cells in the allograft models treated with chemotherapy undergo a dramatic, entinostat-induced (70-100%) conversion to non-proliferative rhabdomyoblasts. Conclusion: Our results suggest that the targeting class I HDACs may provide a therapeutic benefit for selected patients with eRMS. ENT's preclinical in vivo efficacy makes ENT a rational drug candidate in a phase II clinical trial for eRMS.

AB - Background: Rhabdomyosarcoma (RMS) is the most common soft tissue sarcoma in the pediatric cancer population. Survival among metastatic RMS patients has remained dismal yet unimproved for years. We previously identified the class I-specific histone deacetylase inhibitor, entinostat (ENT), as a pharmacological agent that transcriptionally suppresses the PAX3:FOXO1 tumor-initiating fusion gene found in alveolar rhabdomyosarcoma (aRMS), and we further investigated the mechanism by which ENT suppresses PAX3:FOXO1 oncogene and demonstrated the preclinical efficacy of ENT in RMS orthotopic allograft and patient-derived xenograft (PDX) models. In this study, we investigated whether ENT also has antitumor activity in fusion-negative eRMS orthotopic allografts and PDX models either as a single agent or in combination with vincristine (VCR). Methods: We tested the efficacy of ENT and VCR as single agents and in combination in orthotopic allograft and PDX mouse models of eRMS. We then performed CRISPR screening to identify which HDAC among the class I HDACs is responsible for tumor growth inhibition in eRMS. To analyze whether ENT treatment as a single agent or in combination with VCR induces myogenic differentiation, we performed hematoxylin and eosin (H&E) staining in tumors. Results: ENT in combination with the chemotherapy VCR has synergistic antitumor activity in a subset of fusion-negative eRMS in orthotopic "allografts," although PDX mouse models were too hypersensitive to the VCR dose used to detect synergy. Mechanistic studies involving CRISPR suggest that HDAC3 inhibition is the primary mechanism of cell-autonomous cytoreduction in eRMS. Following cytoreduction in vivo, residual tumor cells in the allograft models treated with chemotherapy undergo a dramatic, entinostat-induced (70-100%) conversion to non-proliferative rhabdomyoblasts. Conclusion: Our results suggest that the targeting class I HDACs may provide a therapeutic benefit for selected patients with eRMS. ENT's preclinical in vivo efficacy makes ENT a rational drug candidate in a phase II clinical trial for eRMS.

KW - Embryonal rhabdomyosarcoma (eRMS)

KW - Entinostat

KW - HDAC3

KW - Patient-derived xenograft (PDX)

KW - Vincristine

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