Stemness, Pluripotentiality, and Wnt Antagonism: sFRP4, a Wnt antagonist Mediates Pluripotency and Stemness in Glioblastoma

Gurubharathi Bhuvanalakshmi, Naisarg Gamit, Manasi Patil, Frank Arfuso, Gautam Sethi, Arun Dharmarajan, Alan Prem Kumar, Sudha Warrier

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Background: Chemotherapeutic resistance of glioblastoma has been attributed to a self-renewing subpopulation, the glioma stem cells (GSCs), which is known to be maintained by the Wnt beta-catenin pathway. Our previous findings demonstrated that exogeneous addition of the Wnt antagonist, secreted fizzled-related protein 4 (sFRP4) hampered stem cell properties in GSCs. Methods: To understand the molecular mechanism of sFRP4, we overexpressed sFRP4 (sFRP4 OE) in three human glioblastoma cell lines U87MG, U138MG, and U373MG. We also performed chromatin immunoprecipitation (ChIP) sequencing of sFRP4 OE and RNA sequencing of sFRP4 OE and sFRP4 knocked down U87 cells. Results: We observed nuclear localization of sFRP4, suggesting an unknown nuclear role. ChIP-sequencing of sFRP4 pulldown DNA revealed a homeobox Cphx1, related to the senescence regulator ETS proto-oncogene 2 (ETS2). Furthermore, miRNA885, a p53-mediated apoptosis inducer, was upregulated in sFRP4 OE cells. RNA sequencing analysis suggested that sFRP4-mediated apoptosis is via the Fas-p53 pathway by activating the Wnt calcium and reactive oxygen species pathways. Interestingly, sFRP4 OE cells had decreased stemness, but when knocked down in multipotent mesenchymal stem cells, pluripotentiality was induced and the Wnt beta-catenin pathway was upregulated. Conclusions: This study unveils a novel nuclear role for sFRP4 to promote apoptosis by a possible activation of DNA damage machinery in glioblastoma.

Original languageEnglish
Article number25
Number of pages20
JournalCancers
Volume11
Issue number1
DOIs
Publication statusPublished - Jan 2019

Cite this

Bhuvanalakshmi, Gurubharathi ; Gamit, Naisarg ; Patil, Manasi ; Arfuso, Frank ; Sethi, Gautam ; Dharmarajan, Arun ; Kumar, Alan Prem ; Warrier, Sudha. / Stemness, Pluripotentiality, and Wnt Antagonism : sFRP4, a Wnt antagonist Mediates Pluripotency and Stemness in Glioblastoma. In: Cancers. 2019 ; Vol. 11, No. 1.
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abstract = "Background: Chemotherapeutic resistance of glioblastoma has been attributed to a self-renewing subpopulation, the glioma stem cells (GSCs), which is known to be maintained by the Wnt beta-catenin pathway. Our previous findings demonstrated that exogeneous addition of the Wnt antagonist, secreted fizzled-related protein 4 (sFRP4) hampered stem cell properties in GSCs. Methods: To understand the molecular mechanism of sFRP4, we overexpressed sFRP4 (sFRP4 OE) in three human glioblastoma cell lines U87MG, U138MG, and U373MG. We also performed chromatin immunoprecipitation (ChIP) sequencing of sFRP4 OE and RNA sequencing of sFRP4 OE and sFRP4 knocked down U87 cells. Results: We observed nuclear localization of sFRP4, suggesting an unknown nuclear role. ChIP-sequencing of sFRP4 pulldown DNA revealed a homeobox Cphx1, related to the senescence regulator ETS proto-oncogene 2 (ETS2). Furthermore, miRNA885, a p53-mediated apoptosis inducer, was upregulated in sFRP4 OE cells. RNA sequencing analysis suggested that sFRP4-mediated apoptosis is via the Fas-p53 pathway by activating the Wnt calcium and reactive oxygen species pathways. Interestingly, sFRP4 OE cells had decreased stemness, but when knocked down in multipotent mesenchymal stem cells, pluripotentiality was induced and the Wnt beta-catenin pathway was upregulated. Conclusions: This study unveils a novel nuclear role for sFRP4 to promote apoptosis by a possible activation of DNA damage machinery in glioblastoma.",
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author = "Gurubharathi Bhuvanalakshmi and Naisarg Gamit and Manasi Patil and Frank Arfuso and Gautam Sethi and Arun Dharmarajan and Kumar, {Alan Prem} and Sudha Warrier",
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Stemness, Pluripotentiality, and Wnt Antagonism : sFRP4, a Wnt antagonist Mediates Pluripotency and Stemness in Glioblastoma. / Bhuvanalakshmi, Gurubharathi; Gamit, Naisarg; Patil, Manasi; Arfuso, Frank; Sethi, Gautam; Dharmarajan, Arun; Kumar, Alan Prem; Warrier, Sudha.

In: Cancers, Vol. 11, No. 1, 25, 01.2019.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Stemness, Pluripotentiality, and Wnt Antagonism

T2 - sFRP4, a Wnt antagonist Mediates Pluripotency and Stemness in Glioblastoma

AU - Bhuvanalakshmi, Gurubharathi

AU - Gamit, Naisarg

AU - Patil, Manasi

AU - Arfuso, Frank

AU - Sethi, Gautam

AU - Dharmarajan, Arun

AU - Kumar, Alan Prem

AU - Warrier, Sudha

PY - 2019/1

Y1 - 2019/1

N2 - Background: Chemotherapeutic resistance of glioblastoma has been attributed to a self-renewing subpopulation, the glioma stem cells (GSCs), which is known to be maintained by the Wnt beta-catenin pathway. Our previous findings demonstrated that exogeneous addition of the Wnt antagonist, secreted fizzled-related protein 4 (sFRP4) hampered stem cell properties in GSCs. Methods: To understand the molecular mechanism of sFRP4, we overexpressed sFRP4 (sFRP4 OE) in three human glioblastoma cell lines U87MG, U138MG, and U373MG. We also performed chromatin immunoprecipitation (ChIP) sequencing of sFRP4 OE and RNA sequencing of sFRP4 OE and sFRP4 knocked down U87 cells. Results: We observed nuclear localization of sFRP4, suggesting an unknown nuclear role. ChIP-sequencing of sFRP4 pulldown DNA revealed a homeobox Cphx1, related to the senescence regulator ETS proto-oncogene 2 (ETS2). Furthermore, miRNA885, a p53-mediated apoptosis inducer, was upregulated in sFRP4 OE cells. RNA sequencing analysis suggested that sFRP4-mediated apoptosis is via the Fas-p53 pathway by activating the Wnt calcium and reactive oxygen species pathways. Interestingly, sFRP4 OE cells had decreased stemness, but when knocked down in multipotent mesenchymal stem cells, pluripotentiality was induced and the Wnt beta-catenin pathway was upregulated. Conclusions: This study unveils a novel nuclear role for sFRP4 to promote apoptosis by a possible activation of DNA damage machinery in glioblastoma.

AB - Background: Chemotherapeutic resistance of glioblastoma has been attributed to a self-renewing subpopulation, the glioma stem cells (GSCs), which is known to be maintained by the Wnt beta-catenin pathway. Our previous findings demonstrated that exogeneous addition of the Wnt antagonist, secreted fizzled-related protein 4 (sFRP4) hampered stem cell properties in GSCs. Methods: To understand the molecular mechanism of sFRP4, we overexpressed sFRP4 (sFRP4 OE) in three human glioblastoma cell lines U87MG, U138MG, and U373MG. We also performed chromatin immunoprecipitation (ChIP) sequencing of sFRP4 OE and RNA sequencing of sFRP4 OE and sFRP4 knocked down U87 cells. Results: We observed nuclear localization of sFRP4, suggesting an unknown nuclear role. ChIP-sequencing of sFRP4 pulldown DNA revealed a homeobox Cphx1, related to the senescence regulator ETS proto-oncogene 2 (ETS2). Furthermore, miRNA885, a p53-mediated apoptosis inducer, was upregulated in sFRP4 OE cells. RNA sequencing analysis suggested that sFRP4-mediated apoptosis is via the Fas-p53 pathway by activating the Wnt calcium and reactive oxygen species pathways. Interestingly, sFRP4 OE cells had decreased stemness, but when knocked down in multipotent mesenchymal stem cells, pluripotentiality was induced and the Wnt beta-catenin pathway was upregulated. Conclusions: This study unveils a novel nuclear role for sFRP4 to promote apoptosis by a possible activation of DNA damage machinery in glioblastoma.

KW - Wnt

KW - sFRP4

KW - glioblastoma

KW - pluripotency

KW - p53

KW - EPIGENETIC INACTIVATION

KW - PROTEIN

KW - CELLS

KW - APOPTOSIS

KW - PROLIFERATION

KW - EXPRESSION

KW - KINASE

KW - IDENTIFICATION

KW - REQUIREMENT

KW - ACTIVATION

U2 - 10.3390/cancers11010025

DO - 10.3390/cancers11010025

M3 - Article

VL - 11

JO - Cancers

JF - Cancers

SN - 2072-6694

IS - 1

M1 - 25

ER -