Inhibiting p21-activated kinase induces cell death in vestibular schwannoma and meningioma via mitotic catastrophe

Melania Mercado-Pimentel, Craig Miller, Daniela N. Rolph, Edrick F. Villalobos, Allison M. Dunn, Prithvi M. Mohan, Suzu Igarashi, Xiangdang Liu, Macken Yrun-Duffy, Neal K. Patel, Cecilia M. Read, Ross H. Francis, Adelina Isabella Lane, Swaroop Murugesh, Abraham Jacob

Research output: Contribution to journalArticle

Abstract

Hypothesis: p21-activated kinase (PAK) regulates signaling pathways that promote cell survival and proliferation; therefore, pharmacological inhibition of PAK will induce cell death in vestibular schwannomas (VS) and meningiomas. Background: All VS and many meningiomas result from loss of the neurofibromatosis type 2 (NF2) gene product merlin, with ensuing PAK hyperactivation and increased cell proliferation/survival. Methods: The novel small molecule PAK inhibitors PI-8 and PI-15-tested in schwannoma and meningioma cells-perturb molecular signaling and induce cell death. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, flow cytometry, and terminal deoxynucleotidyl transferase dUTP nick end labeling assay analyzed PAK inhibitors' effect on cell viability, cell cycle, and cell death, respectively. Western blots evaluated activation and expression of cell proliferation, apoptotic, and mitotic catastrophe markers. Light microscopy evaluated cell morphology, and immunocytochemistry analyzed cellular localization of phospho-Merlin and autophagy-related protein. Results: Treatment with PI-8 and PI-15 decreased cell viability at 0.65 to 3.7mM 50% inhibitory concentration (IC50) in schwannoma and meningioma cells. Terminal deoxynucleotidyl transferase dUTP nick end labeling and immunocytochemistry studies show that PI-8 and PI-15 induce mitotic catastrophe but not apoptosis in HEI193 cells while in BenMen1 cells, PI-8 induces autophagy and mitotic catastrophe. PI-15 induces apoptosis in BenMen1 cells. PAK inhibitor treated cells show phospho-Merlin localized to over-duplicated centrosomes of dividing cells, multiple enlarged nuclei, and misaligned/missegregated chromosomes-markers for mitotic catastrophe. Increased autophagy-related protein levels in the nucleus confirmed this cell death type. PI-8 and PI-15 inhibits PAK in both cell lines. However, only PI-15 inhibits v-akt murine thymoma viral oncogene homolog in BenMen1 cells. Conclusion: PAK inhibitors induce cell death in schwannoma and meningioma cells, at least in part, by mitotic catastrophe.

Original languageEnglish (US)
Pages (from-to)139-146
Number of pages8
JournalOtology and Neurotology
Volume38
Issue number1
DOIs
StatePublished - 2017

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p21-Activated Kinases
Acoustic Neuroma
Meningioma
Cell Death
Neurofibromin 2
Cell Survival
Neurilemmoma
DNA Nucleotidylexotransferase
Cell Proliferation
Inhibitory Concentration 50
Immunohistochemistry
Apoptosis
Neurofibromatosis 2
Centrosome
Thymoma
Autophagy
Genetic Markers
Oncogenes
Microscopy
Cell Cycle

Keywords

  • Apoptosis
  • Autophagy
  • Meningioma
  • Mitotic catastrophe
  • Neurofibromatosis type 2
  • p21-activated kinase (PAK)
  • Vestibular schwannoma

ASJC Scopus subject areas

  • Otorhinolaryngology
  • Sensory Systems
  • Clinical Neurology

Cite this

Inhibiting p21-activated kinase induces cell death in vestibular schwannoma and meningioma via mitotic catastrophe. / Mercado-Pimentel, Melania; Miller, Craig; Rolph, Daniela N.; Villalobos, Edrick F.; Dunn, Allison M.; Mohan, Prithvi M.; Igarashi, Suzu; Liu, Xiangdang; Yrun-Duffy, Macken; Patel, Neal K.; Read, Cecilia M.; Francis, Ross H.; Lane, Adelina Isabella; Murugesh, Swaroop; Jacob, Abraham.

In: Otology and Neurotology, Vol. 38, No. 1, 2017, p. 139-146.

Research output: Contribution to journalArticle

Mercado-Pimentel, M, Miller, C, Rolph, DN, Villalobos, EF, Dunn, AM, Mohan, PM, Igarashi, S, Liu, X, Yrun-Duffy, M, Patel, NK, Read, CM, Francis, RH, Lane, AI, Murugesh, S & Jacob, A 2017, 'Inhibiting p21-activated kinase induces cell death in vestibular schwannoma and meningioma via mitotic catastrophe', Otology and Neurotology, vol. 38, no. 1, pp. 139-146. https://doi.org/10.1097/MAO.0000000000001247
Mercado-Pimentel, Melania ; Miller, Craig ; Rolph, Daniela N. ; Villalobos, Edrick F. ; Dunn, Allison M. ; Mohan, Prithvi M. ; Igarashi, Suzu ; Liu, Xiangdang ; Yrun-Duffy, Macken ; Patel, Neal K. ; Read, Cecilia M. ; Francis, Ross H. ; Lane, Adelina Isabella ; Murugesh, Swaroop ; Jacob, Abraham. / Inhibiting p21-activated kinase induces cell death in vestibular schwannoma and meningioma via mitotic catastrophe. In: Otology and Neurotology. 2017 ; Vol. 38, No. 1. pp. 139-146.
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abstract = "Hypothesis: p21-activated kinase (PAK) regulates signaling pathways that promote cell survival and proliferation; therefore, pharmacological inhibition of PAK will induce cell death in vestibular schwannomas (VS) and meningiomas. Background: All VS and many meningiomas result from loss of the neurofibromatosis type 2 (NF2) gene product merlin, with ensuing PAK hyperactivation and increased cell proliferation/survival. Methods: The novel small molecule PAK inhibitors PI-8 and PI-15-tested in schwannoma and meningioma cells-perturb molecular signaling and induce cell death. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, flow cytometry, and terminal deoxynucleotidyl transferase dUTP nick end labeling assay analyzed PAK inhibitors' effect on cell viability, cell cycle, and cell death, respectively. Western blots evaluated activation and expression of cell proliferation, apoptotic, and mitotic catastrophe markers. Light microscopy evaluated cell morphology, and immunocytochemistry analyzed cellular localization of phospho-Merlin and autophagy-related protein. Results: Treatment with PI-8 and PI-15 decreased cell viability at 0.65 to 3.7mM 50{\%} inhibitory concentration (IC50) in schwannoma and meningioma cells. Terminal deoxynucleotidyl transferase dUTP nick end labeling and immunocytochemistry studies show that PI-8 and PI-15 induce mitotic catastrophe but not apoptosis in HEI193 cells while in BenMen1 cells, PI-8 induces autophagy and mitotic catastrophe. PI-15 induces apoptosis in BenMen1 cells. PAK inhibitor treated cells show phospho-Merlin localized to over-duplicated centrosomes of dividing cells, multiple enlarged nuclei, and misaligned/missegregated chromosomes-markers for mitotic catastrophe. Increased autophagy-related protein levels in the nucleus confirmed this cell death type. PI-8 and PI-15 inhibits PAK in both cell lines. However, only PI-15 inhibits v-akt murine thymoma viral oncogene homolog in BenMen1 cells. Conclusion: PAK inhibitors induce cell death in schwannoma and meningioma cells, at least in part, by mitotic catastrophe.",
keywords = "Apoptosis, Autophagy, Meningioma, Mitotic catastrophe, Neurofibromatosis type 2, p21-activated kinase (PAK), Vestibular schwannoma",
author = "Melania Mercado-Pimentel and Craig Miller and Rolph, {Daniela N.} and Villalobos, {Edrick F.} and Dunn, {Allison M.} and Mohan, {Prithvi M.} and Suzu Igarashi and Xiangdang Liu and Macken Yrun-Duffy and Patel, {Neal K.} and Read, {Cecilia M.} and Francis, {Ross H.} and Lane, {Adelina Isabella} and Swaroop Murugesh and Abraham Jacob",
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T1 - Inhibiting p21-activated kinase induces cell death in vestibular schwannoma and meningioma via mitotic catastrophe

AU - Mercado-Pimentel, Melania

AU - Miller, Craig

AU - Rolph, Daniela N.

AU - Villalobos, Edrick F.

AU - Dunn, Allison M.

AU - Mohan, Prithvi M.

AU - Igarashi, Suzu

AU - Liu, Xiangdang

AU - Yrun-Duffy, Macken

AU - Patel, Neal K.

AU - Read, Cecilia M.

AU - Francis, Ross H.

AU - Lane, Adelina Isabella

AU - Murugesh, Swaroop

AU - Jacob, Abraham

PY - 2017

Y1 - 2017

N2 - Hypothesis: p21-activated kinase (PAK) regulates signaling pathways that promote cell survival and proliferation; therefore, pharmacological inhibition of PAK will induce cell death in vestibular schwannomas (VS) and meningiomas. Background: All VS and many meningiomas result from loss of the neurofibromatosis type 2 (NF2) gene product merlin, with ensuing PAK hyperactivation and increased cell proliferation/survival. Methods: The novel small molecule PAK inhibitors PI-8 and PI-15-tested in schwannoma and meningioma cells-perturb molecular signaling and induce cell death. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, flow cytometry, and terminal deoxynucleotidyl transferase dUTP nick end labeling assay analyzed PAK inhibitors' effect on cell viability, cell cycle, and cell death, respectively. Western blots evaluated activation and expression of cell proliferation, apoptotic, and mitotic catastrophe markers. Light microscopy evaluated cell morphology, and immunocytochemistry analyzed cellular localization of phospho-Merlin and autophagy-related protein. Results: Treatment with PI-8 and PI-15 decreased cell viability at 0.65 to 3.7mM 50% inhibitory concentration (IC50) in schwannoma and meningioma cells. Terminal deoxynucleotidyl transferase dUTP nick end labeling and immunocytochemistry studies show that PI-8 and PI-15 induce mitotic catastrophe but not apoptosis in HEI193 cells while in BenMen1 cells, PI-8 induces autophagy and mitotic catastrophe. PI-15 induces apoptosis in BenMen1 cells. PAK inhibitor treated cells show phospho-Merlin localized to over-duplicated centrosomes of dividing cells, multiple enlarged nuclei, and misaligned/missegregated chromosomes-markers for mitotic catastrophe. Increased autophagy-related protein levels in the nucleus confirmed this cell death type. PI-8 and PI-15 inhibits PAK in both cell lines. However, only PI-15 inhibits v-akt murine thymoma viral oncogene homolog in BenMen1 cells. Conclusion: PAK inhibitors induce cell death in schwannoma and meningioma cells, at least in part, by mitotic catastrophe.

AB - Hypothesis: p21-activated kinase (PAK) regulates signaling pathways that promote cell survival and proliferation; therefore, pharmacological inhibition of PAK will induce cell death in vestibular schwannomas (VS) and meningiomas. Background: All VS and many meningiomas result from loss of the neurofibromatosis type 2 (NF2) gene product merlin, with ensuing PAK hyperactivation and increased cell proliferation/survival. Methods: The novel small molecule PAK inhibitors PI-8 and PI-15-tested in schwannoma and meningioma cells-perturb molecular signaling and induce cell death. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, flow cytometry, and terminal deoxynucleotidyl transferase dUTP nick end labeling assay analyzed PAK inhibitors' effect on cell viability, cell cycle, and cell death, respectively. Western blots evaluated activation and expression of cell proliferation, apoptotic, and mitotic catastrophe markers. Light microscopy evaluated cell morphology, and immunocytochemistry analyzed cellular localization of phospho-Merlin and autophagy-related protein. Results: Treatment with PI-8 and PI-15 decreased cell viability at 0.65 to 3.7mM 50% inhibitory concentration (IC50) in schwannoma and meningioma cells. Terminal deoxynucleotidyl transferase dUTP nick end labeling and immunocytochemistry studies show that PI-8 and PI-15 induce mitotic catastrophe but not apoptosis in HEI193 cells while in BenMen1 cells, PI-8 induces autophagy and mitotic catastrophe. PI-15 induces apoptosis in BenMen1 cells. PAK inhibitor treated cells show phospho-Merlin localized to over-duplicated centrosomes of dividing cells, multiple enlarged nuclei, and misaligned/missegregated chromosomes-markers for mitotic catastrophe. Increased autophagy-related protein levels in the nucleus confirmed this cell death type. PI-8 and PI-15 inhibits PAK in both cell lines. However, only PI-15 inhibits v-akt murine thymoma viral oncogene homolog in BenMen1 cells. Conclusion: PAK inhibitors induce cell death in schwannoma and meningioma cells, at least in part, by mitotic catastrophe.

KW - Apoptosis

KW - Autophagy

KW - Meningioma

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KW - Neurofibromatosis type 2

KW - p21-activated kinase (PAK)

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