Identification of PADI2 as a potential breast cancer biomarker and therapeutic target

John L. McElwee, Sunish Mohanan, Obi L. Griffith, Heike C. Breuer, Lynne J. Anguish, Brian D. Cherrington, Ashley M. Palmer, Louise R. Howe, Venkataraman Subramanian, Corey P. Causey, Paul R. Thompson, Joe Gray, Scott A. Coonrod

Research output: Contribution to journalArticle

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Abstract

Background: We have recently reported that the expression of peptidylarginine deiminase 2 (PADI2) is regulated by EGF in mammary cancer cells and appears to play a role in the proliferation of normal mammary epithelium; however, the role of PADI2 in the pathogenesis of human breast cancer has yet to be investigated. Thus, the goals of this study were to examine whether PADI2 plays a role in mammary tumor progression, and whether the inhibition of PADI activity has anti-tumor effects.Methods: RNA-seq data from a collection of 57 breast cancer cell lines was queried for PADI2 levels, and correlations with known subtype and HER2/ERBB2 status were evaluated. To examine PADI2 expression levels during breast cancer progression, the cell lines from the MCF10AT model were used. The efficacy of the PADI inhibitor, Cl-amidine, was tested in vitro using MCF10DCIS cells grown in 2D-monolayers and 3D-spheroids, and in vivo using MCF10DCIS tumor xenografts. Treated MCF10DCIS cells were examined by flow-cytometry to determine the extent of apoptosis and by RT2 Profiler PCR Cell Cycle Array to detect alterations in cell cycle associated genes.Results: We show by RNA-seq that PADI2 mRNA expression is highly correlated with HER2/ERBB2 (p = 2.2 × 106) in luminal breast cancer cell lines. Using the MCF10AT model of breast cancer progression, we then demonstrate that PADI2 expression increases during the transition of normal mammary epithelium to fully malignant breast carcinomas, with a strong peak of PADI2 expression and activity being observed in the MCF10DCIS cell line, which models human comedo-DCIS lesions. Next, we show that a PADI inhibitor, Cl-amidine, strongly suppresses the growth of MCF10DCIS monolayers and tumor spheroids in culture. We then carried out preclinical studies in nude (nu/nu) mice and found that Cl-amidine also suppressed the growth of xenografted MCF10DCIS tumors by more than 3-fold. Lastly, we performed cell cycle array analysis of Cl-amidine treated and control MCF10DCIS cells, and found that the PADI inhibitor strongly affects the expression of several cell cycle genes implicated in tumor progression, including p21, GADD45α, and Ki67.Conclusion: Together, these results suggest that PADI2 may function as an important new biomarker for HER2/ERBB2+ tumors and that Cl-amidine represents a new candidate for breast cancer therapy.

Original languageEnglish (US)
Article number500
JournalBMC Cancer
Volume12
DOIs
StatePublished - Oct 30 2012

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Tumor Biomarkers
Breast Neoplasms
Cell Line
cdc Genes
Neoplasms
Therapeutics
Cell Cycle
Breast
Epithelium
Tissue Array Analysis
RNA
protein-arginine deiminase
Carcinoma, Intraductal, Noninfiltrating
Growth
Epidermal Growth Factor
Heterografts
Flow Cytometry
Biomarkers
N-alpha-benzoyl-N5-(2-chloro-1-iminoethyl)-L-ornithine amide
Apoptosis

Keywords

  • Breast cancer
  • Citrullination
  • Cl-amidine
  • HER2/ERBB2
  • Luminal
  • PAD2/PADI2
  • Peptidylarginine deiminase

ASJC Scopus subject areas

  • Oncology
  • Cancer Research
  • Genetics

Cite this

McElwee, J. L., Mohanan, S., Griffith, O. L., Breuer, H. C., Anguish, L. J., Cherrington, B. D., ... Coonrod, S. A. (2012). Identification of PADI2 as a potential breast cancer biomarker and therapeutic target. BMC Cancer, 12, [500]. https://doi.org/10.1186/1471-2407-12-500

Identification of PADI2 as a potential breast cancer biomarker and therapeutic target. / McElwee, John L.; Mohanan, Sunish; Griffith, Obi L.; Breuer, Heike C.; Anguish, Lynne J.; Cherrington, Brian D.; Palmer, Ashley M.; Howe, Louise R.; Subramanian, Venkataraman; Causey, Corey P.; Thompson, Paul R.; Gray, Joe; Coonrod, Scott A.

In: BMC Cancer, Vol. 12, 500, 30.10.2012.

Research output: Contribution to journalArticle

McElwee, JL, Mohanan, S, Griffith, OL, Breuer, HC, Anguish, LJ, Cherrington, BD, Palmer, AM, Howe, LR, Subramanian, V, Causey, CP, Thompson, PR, Gray, J & Coonrod, SA 2012, 'Identification of PADI2 as a potential breast cancer biomarker and therapeutic target', BMC Cancer, vol. 12, 500. https://doi.org/10.1186/1471-2407-12-500
McElwee JL, Mohanan S, Griffith OL, Breuer HC, Anguish LJ, Cherrington BD et al. Identification of PADI2 as a potential breast cancer biomarker and therapeutic target. BMC Cancer. 2012 Oct 30;12. 500. https://doi.org/10.1186/1471-2407-12-500
McElwee, John L. ; Mohanan, Sunish ; Griffith, Obi L. ; Breuer, Heike C. ; Anguish, Lynne J. ; Cherrington, Brian D. ; Palmer, Ashley M. ; Howe, Louise R. ; Subramanian, Venkataraman ; Causey, Corey P. ; Thompson, Paul R. ; Gray, Joe ; Coonrod, Scott A. / Identification of PADI2 as a potential breast cancer biomarker and therapeutic target. In: BMC Cancer. 2012 ; Vol. 12.
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abstract = "Background: We have recently reported that the expression of peptidylarginine deiminase 2 (PADI2) is regulated by EGF in mammary cancer cells and appears to play a role in the proliferation of normal mammary epithelium; however, the role of PADI2 in the pathogenesis of human breast cancer has yet to be investigated. Thus, the goals of this study were to examine whether PADI2 plays a role in mammary tumor progression, and whether the inhibition of PADI activity has anti-tumor effects.Methods: RNA-seq data from a collection of 57 breast cancer cell lines was queried for PADI2 levels, and correlations with known subtype and HER2/ERBB2 status were evaluated. To examine PADI2 expression levels during breast cancer progression, the cell lines from the MCF10AT model were used. The efficacy of the PADI inhibitor, Cl-amidine, was tested in vitro using MCF10DCIS cells grown in 2D-monolayers and 3D-spheroids, and in vivo using MCF10DCIS tumor xenografts. Treated MCF10DCIS cells were examined by flow-cytometry to determine the extent of apoptosis and by RT2 Profiler PCR Cell Cycle Array to detect alterations in cell cycle associated genes.Results: We show by RNA-seq that PADI2 mRNA expression is highly correlated with HER2/ERBB2 (p = 2.2 × 106) in luminal breast cancer cell lines. Using the MCF10AT model of breast cancer progression, we then demonstrate that PADI2 expression increases during the transition of normal mammary epithelium to fully malignant breast carcinomas, with a strong peak of PADI2 expression and activity being observed in the MCF10DCIS cell line, which models human comedo-DCIS lesions. Next, we show that a PADI inhibitor, Cl-amidine, strongly suppresses the growth of MCF10DCIS monolayers and tumor spheroids in culture. We then carried out preclinical studies in nude (nu/nu) mice and found that Cl-amidine also suppressed the growth of xenografted MCF10DCIS tumors by more than 3-fold. Lastly, we performed cell cycle array analysis of Cl-amidine treated and control MCF10DCIS cells, and found that the PADI inhibitor strongly affects the expression of several cell cycle genes implicated in tumor progression, including p21, GADD45α, and Ki67.Conclusion: Together, these results suggest that PADI2 may function as an important new biomarker for HER2/ERBB2+ tumors and that Cl-amidine represents a new candidate for breast cancer therapy.",
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author = "McElwee, {John L.} and Sunish Mohanan and Griffith, {Obi L.} and Breuer, {Heike C.} and Anguish, {Lynne J.} and Cherrington, {Brian D.} and Palmer, {Ashley M.} and Howe, {Louise R.} and Venkataraman Subramanian and Causey, {Corey P.} and Thompson, {Paul R.} and Joe Gray and Coonrod, {Scott A.}",
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T1 - Identification of PADI2 as a potential breast cancer biomarker and therapeutic target

AU - McElwee, John L.

AU - Mohanan, Sunish

AU - Griffith, Obi L.

AU - Breuer, Heike C.

AU - Anguish, Lynne J.

AU - Cherrington, Brian D.

AU - Palmer, Ashley M.

AU - Howe, Louise R.

AU - Subramanian, Venkataraman

AU - Causey, Corey P.

AU - Thompson, Paul R.

AU - Gray, Joe

AU - Coonrod, Scott A.

PY - 2012/10/30

Y1 - 2012/10/30

N2 - Background: We have recently reported that the expression of peptidylarginine deiminase 2 (PADI2) is regulated by EGF in mammary cancer cells and appears to play a role in the proliferation of normal mammary epithelium; however, the role of PADI2 in the pathogenesis of human breast cancer has yet to be investigated. Thus, the goals of this study were to examine whether PADI2 plays a role in mammary tumor progression, and whether the inhibition of PADI activity has anti-tumor effects.Methods: RNA-seq data from a collection of 57 breast cancer cell lines was queried for PADI2 levels, and correlations with known subtype and HER2/ERBB2 status were evaluated. To examine PADI2 expression levels during breast cancer progression, the cell lines from the MCF10AT model were used. The efficacy of the PADI inhibitor, Cl-amidine, was tested in vitro using MCF10DCIS cells grown in 2D-monolayers and 3D-spheroids, and in vivo using MCF10DCIS tumor xenografts. Treated MCF10DCIS cells were examined by flow-cytometry to determine the extent of apoptosis and by RT2 Profiler PCR Cell Cycle Array to detect alterations in cell cycle associated genes.Results: We show by RNA-seq that PADI2 mRNA expression is highly correlated with HER2/ERBB2 (p = 2.2 × 106) in luminal breast cancer cell lines. Using the MCF10AT model of breast cancer progression, we then demonstrate that PADI2 expression increases during the transition of normal mammary epithelium to fully malignant breast carcinomas, with a strong peak of PADI2 expression and activity being observed in the MCF10DCIS cell line, which models human comedo-DCIS lesions. Next, we show that a PADI inhibitor, Cl-amidine, strongly suppresses the growth of MCF10DCIS monolayers and tumor spheroids in culture. We then carried out preclinical studies in nude (nu/nu) mice and found that Cl-amidine also suppressed the growth of xenografted MCF10DCIS tumors by more than 3-fold. Lastly, we performed cell cycle array analysis of Cl-amidine treated and control MCF10DCIS cells, and found that the PADI inhibitor strongly affects the expression of several cell cycle genes implicated in tumor progression, including p21, GADD45α, and Ki67.Conclusion: Together, these results suggest that PADI2 may function as an important new biomarker for HER2/ERBB2+ tumors and that Cl-amidine represents a new candidate for breast cancer therapy.

AB - Background: We have recently reported that the expression of peptidylarginine deiminase 2 (PADI2) is regulated by EGF in mammary cancer cells and appears to play a role in the proliferation of normal mammary epithelium; however, the role of PADI2 in the pathogenesis of human breast cancer has yet to be investigated. Thus, the goals of this study were to examine whether PADI2 plays a role in mammary tumor progression, and whether the inhibition of PADI activity has anti-tumor effects.Methods: RNA-seq data from a collection of 57 breast cancer cell lines was queried for PADI2 levels, and correlations with known subtype and HER2/ERBB2 status were evaluated. To examine PADI2 expression levels during breast cancer progression, the cell lines from the MCF10AT model were used. The efficacy of the PADI inhibitor, Cl-amidine, was tested in vitro using MCF10DCIS cells grown in 2D-monolayers and 3D-spheroids, and in vivo using MCF10DCIS tumor xenografts. Treated MCF10DCIS cells were examined by flow-cytometry to determine the extent of apoptosis and by RT2 Profiler PCR Cell Cycle Array to detect alterations in cell cycle associated genes.Results: We show by RNA-seq that PADI2 mRNA expression is highly correlated with HER2/ERBB2 (p = 2.2 × 106) in luminal breast cancer cell lines. Using the MCF10AT model of breast cancer progression, we then demonstrate that PADI2 expression increases during the transition of normal mammary epithelium to fully malignant breast carcinomas, with a strong peak of PADI2 expression and activity being observed in the MCF10DCIS cell line, which models human comedo-DCIS lesions. Next, we show that a PADI inhibitor, Cl-amidine, strongly suppresses the growth of MCF10DCIS monolayers and tumor spheroids in culture. We then carried out preclinical studies in nude (nu/nu) mice and found that Cl-amidine also suppressed the growth of xenografted MCF10DCIS tumors by more than 3-fold. Lastly, we performed cell cycle array analysis of Cl-amidine treated and control MCF10DCIS cells, and found that the PADI inhibitor strongly affects the expression of several cell cycle genes implicated in tumor progression, including p21, GADD45α, and Ki67.Conclusion: Together, these results suggest that PADI2 may function as an important new biomarker for HER2/ERBB2+ tumors and that Cl-amidine represents a new candidate for breast cancer therapy.

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KW - Citrullination

KW - Cl-amidine

KW - HER2/ERBB2

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KW - PAD2/PADI2

KW - Peptidylarginine deiminase

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