ACVR1C/SMAD2 signaling promotes invasion and growth in retinoblastoma

Laura Asnaghi, David T. White, Nolan Key, Joshua Choi, Alka Mahale, Hind Alkatan, Deepak P. Edward, Sahar M. Elkhamary, Saleh Al-Mesfer, Azza Maktabi, Christopher G. Hurtado, Grace Y. Lee, Angel M. Carcaboso, Jeffrey Mumm, Leen Abu Safieh, Charles G Eberhart

Research output: Contribution to journalArticle

Abstract

Retinoblastoma is the most common intraocular cancer in children. While the primary tumor can often be treated by local or systemic chemotherapy, metastatic dissemination is generally resistant to therapy and remains a leading cause of pediatric cancer death in much of the world. In order to identify new therapeutic targets in aggressive tumors, we sequenced RNA transcripts in five snap frozen retinoblastomas which invaded the optic nerve and five which did not. A three-fold increase was noted in mRNA levels of ACVR1C/ALK7, a type I receptor of the TGF-β family, in invasive retinoblastomas, while downregulation of DACT2 and LEFTY2, negative modulators of the ACVR1C signaling, was observed in most invasive tumors. A two- to three-fold increase in ACVR1C mRNA was also found in invasive WERI Rb1 and Y79 cells as compared to non-invasive cells in vitro. Transcripts of ACVR1C receptor and its ligands (Nodal, Activin A/B, and GDF3) were expressed in six retinoblastoma lines, and evidence of downstream SMAD2 signaling was present in all these lines. Pharmacological inhibition of ACVR1C signaling using SB505124, or genetic downregulation of the receptor using shRNA potently suppressed invasion, growth, survival, and reduced the protein levels of the mesenchymal markers ZEB1 and Snail. The inhibitory effects on invasion, growth, and proliferation were recapitulated by knocking down SMAD2, but not SMAD3. Finally, in an orthotopic zebrafish model of retinoblastoma, a 55% decrease in tumor spread was noted (p = 0.0026) when larvae were treated with 3 µM of SB505124, as compared to DMSO. Similarly, knockdown of ACVR1C in injected tumor cells using shRNA also resulted in a 54% reduction in tumor dissemination in the zebrafish eye as compared to scrambled shRNA control (p = 0.0005). Our data support a role for the ACVR1C/SMAD2 pathway in promoting invasion and growth of retinoblastoma.

Original languageEnglish (US)
JournalOncogene
DOIs
StateAccepted/In press - Jan 1 2018

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Retinoblastoma
Growth
Neoplasms
Small Interfering RNA
Zebrafish
Down-Regulation
Messenger RNA
Optic Nerve
Dimethyl Sulfoxide
Larva
Pharmacology
RNA
Pediatrics
Ligands
Drug Therapy
Therapeutics

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cancer Research

Cite this

ACVR1C/SMAD2 signaling promotes invasion and growth in retinoblastoma. / Asnaghi, Laura; White, David T.; Key, Nolan; Choi, Joshua; Mahale, Alka; Alkatan, Hind; Edward, Deepak P.; Elkhamary, Sahar M.; Al-Mesfer, Saleh; Maktabi, Azza; Hurtado, Christopher G.; Lee, Grace Y.; Carcaboso, Angel M.; Mumm, Jeffrey; Safieh, Leen Abu; Eberhart, Charles G.

In: Oncogene, 01.01.2018.

Research output: Contribution to journalArticle

Asnaghi, L, White, DT, Key, N, Choi, J, Mahale, A, Alkatan, H, Edward, DP, Elkhamary, SM, Al-Mesfer, S, Maktabi, A, Hurtado, CG, Lee, GY, Carcaboso, AM, Mumm, J, Safieh, LA & Eberhart, CG 2018, 'ACVR1C/SMAD2 signaling promotes invasion and growth in retinoblastoma', Oncogene. https://doi.org/10.1038/s41388-018-0543-2
Asnaghi, Laura ; White, David T. ; Key, Nolan ; Choi, Joshua ; Mahale, Alka ; Alkatan, Hind ; Edward, Deepak P. ; Elkhamary, Sahar M. ; Al-Mesfer, Saleh ; Maktabi, Azza ; Hurtado, Christopher G. ; Lee, Grace Y. ; Carcaboso, Angel M. ; Mumm, Jeffrey ; Safieh, Leen Abu ; Eberhart, Charles G. / ACVR1C/SMAD2 signaling promotes invasion and growth in retinoblastoma. In: Oncogene. 2018.
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abstract = "Retinoblastoma is the most common intraocular cancer in children. While the primary tumor can often be treated by local or systemic chemotherapy, metastatic dissemination is generally resistant to therapy and remains a leading cause of pediatric cancer death in much of the world. In order to identify new therapeutic targets in aggressive tumors, we sequenced RNA transcripts in five snap frozen retinoblastomas which invaded the optic nerve and five which did not. A three-fold increase was noted in mRNA levels of ACVR1C/ALK7, a type I receptor of the TGF-β family, in invasive retinoblastomas, while downregulation of DACT2 and LEFTY2, negative modulators of the ACVR1C signaling, was observed in most invasive tumors. A two- to three-fold increase in ACVR1C mRNA was also found in invasive WERI Rb1 and Y79 cells as compared to non-invasive cells in vitro. Transcripts of ACVR1C receptor and its ligands (Nodal, Activin A/B, and GDF3) were expressed in six retinoblastoma lines, and evidence of downstream SMAD2 signaling was present in all these lines. Pharmacological inhibition of ACVR1C signaling using SB505124, or genetic downregulation of the receptor using shRNA potently suppressed invasion, growth, survival, and reduced the protein levels of the mesenchymal markers ZEB1 and Snail. The inhibitory effects on invasion, growth, and proliferation were recapitulated by knocking down SMAD2, but not SMAD3. Finally, in an orthotopic zebrafish model of retinoblastoma, a 55{\%} decrease in tumor spread was noted (p = 0.0026) when larvae were treated with 3 µM of SB505124, as compared to DMSO. Similarly, knockdown of ACVR1C in injected tumor cells using shRNA also resulted in a 54{\%} reduction in tumor dissemination in the zebrafish eye as compared to scrambled shRNA control (p = 0.0005). Our data support a role for the ACVR1C/SMAD2 pathway in promoting invasion and growth of retinoblastoma.",
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AU - Mahale, Alka

AU - Alkatan, Hind

AU - Edward, Deepak P.

AU - Elkhamary, Sahar M.

AU - Al-Mesfer, Saleh

AU - Maktabi, Azza

AU - Hurtado, Christopher G.

AU - Lee, Grace Y.

AU - Carcaboso, Angel M.

AU - Mumm, Jeffrey

AU - Safieh, Leen Abu

AU - Eberhart, Charles G

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