Haploinsufficient tumor suppressor PRP4K is negatively regulated during epithelial-to-mesenchymal transition

Livia E. Clarke; Allyson Cook; Sabateeshan Mathavarajah; Amit Bera; Jayme Salsman; Elias Habib; Carter Van Iderstine; Moamen Bydoun; Stephen M. Lewis; Graham Dellaire

doi:10.1096/fj.202001063R

Haploinsufficient tumor suppressor PRP4K is negatively regulated during epithelial-to-mesenchymal transition

Livia E. Clarke, Allyson Cook, Sabateeshan Mathavarajah, Amit Bera, Jayme Salsman, Elias Habib, Carter Van Iderstine, Moamen Bydoun, Stephen M. Lewis, Graham Dellaire

Research output: Contribution to journal › Article › peer-review

3 Citations (Scopus)

Abstract

The pre-mRNA processing factor 4 kinase (PRP4K, also known as PRPF4B) is an essential gene. However, reduced PRP4K expression is associated with aggressive breast and ovarian cancer phenotypes including taxane therapy resistance, increased cell migration and invasion in vitro, and cancer metastasis in mice. These results are consistent with PRP4K being a haploinsufficient tumor suppressor. Increased cell migration and invasion is associated with epithelial-to-mesenchymal transition (EMT), but how reduced PRP4K levels affect normal epithelial cell migration or EMT has not been studied. Depletion of PRP4K by small hairpin RNA (shRNA) in non-transformed mammary epithelial cell lines (MCF10A, HMLE) reduced or had no effect on 2D migration in the scratch assay but resulted in greater invasive potential in 3D transwell assays. Depletion of PRP4K in mesenchymal triple-negative breast cancer cells (MDA-MB-231) resulted in both enhanced 2D migration and 3D invasion, with 3D invasion correlated with higher fibronectin levels in both MDA-MB-231 and MCF10A cells and without changes in E-cadherin. Induction of EMT in MCF10A cells, by treatment with WNT-5a and TGF-β1, or depletion of eukaryotic translation initiation factor 3e (eIF3e) by shRNA, resulted in significantly reduced PRP4K expression. Mechanistically, induction of EMT by WNT-5a/TGF-β1 reduced PRP4K transcript levels, whereas eIF3e depletion led to reduced PRP4K translation. Finally, reduced PRP4K levels after eIF3e depletion correlated with increased YAP activity and nuclear localization, both of which are reversed by overexpression of exogenous PRP4K. Thus, PRP4K is a haploinsufficient tumor suppressor negatively regulated by EMT, that when depleted in normal mammary cells can increase cell invasion without inducing full EMT.

Original language	English
Article number	e22001
Journal	FASEB Journal
Volume	35
Issue number	11
DOIs	https://doi.org/10.1096/fj.202001063R
Publication status	Published - Nov 2021

Bibliographical note

Funding Information:
This work was funded by a Breast Cancer Society/QEII Foundation operating grant awarded to GD and an Operating Grant (#24135) from the Cancer Research Society and the Beatrice Hunter Cancer Research Institute (BHCRI) awarded to SML. GD and SML are Senior Scientists of the BHCRI, and both AC and LEC were supported by Cancer Research Training Program awards from the BHCRI with funds provided by CBCF—Atlantic, The Canadian Cancer Society, Nova Scotia Division as part of The Terry Fox Foundation Strategic Health Research Training (STIHR) Program in Cancer Research at the Canadian Institutes of Health Research (CIHR). SM is supported by a CGSM studentship from Natural Sciences and Engineering Research Council of Canada (NSERC), a Predoctoral Fellowship from the Killam Trusts, and a Scotia Scholar award from the Nova Scotia Health Research Foundation.

Publisher Copyright:
© 2021 The Authors. The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology.

ASJC Scopus Subject Areas

Biotechnology
Biochemistry
Molecular Biology
Genetics

PubMed: MeSH publication types

Journal Article
Research Support, Non-U.S. Gov't

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1096/fj.202001063R

Cite this

@article{6d167e3a7478453e83bae69dcf8f8dfc,

title = "Haploinsufficient tumor suppressor PRP4K is negatively regulated during epithelial-to-mesenchymal transition",

abstract = "The pre-mRNA processing factor 4 kinase (PRP4K, also known as PRPF4B) is an essential gene. However, reduced PRP4K expression is associated with aggressive breast and ovarian cancer phenotypes including taxane therapy resistance, increased cell migration and invasion in vitro, and cancer metastasis in mice. These results are consistent with PRP4K being a haploinsufficient tumor suppressor. Increased cell migration and invasion is associated with epithelial-to-mesenchymal transition (EMT), but how reduced PRP4K levels affect normal epithelial cell migration or EMT has not been studied. Depletion of PRP4K by small hairpin RNA (shRNA) in non-transformed mammary epithelial cell lines (MCF10A, HMLE) reduced or had no effect on 2D migration in the scratch assay but resulted in greater invasive potential in 3D transwell assays. Depletion of PRP4K in mesenchymal triple-negative breast cancer cells (MDA-MB-231) resulted in both enhanced 2D migration and 3D invasion, with 3D invasion correlated with higher fibronectin levels in both MDA-MB-231 and MCF10A cells and without changes in E-cadherin. Induction of EMT in MCF10A cells, by treatment with WNT-5a and TGF-β1, or depletion of eukaryotic translation initiation factor 3e (eIF3e) by shRNA, resulted in significantly reduced PRP4K expression. Mechanistically, induction of EMT by WNT-5a/TGF-β1 reduced PRP4K transcript levels, whereas eIF3e depletion led to reduced PRP4K translation. Finally, reduced PRP4K levels after eIF3e depletion correlated with increased YAP activity and nuclear localization, both of which are reversed by overexpression of exogenous PRP4K. Thus, PRP4K is a haploinsufficient tumor suppressor negatively regulated by EMT, that when depleted in normal mammary cells can increase cell invasion without inducing full EMT.",

author = "Clarke, {Livia E.} and Allyson Cook and Sabateeshan Mathavarajah and Amit Bera and Jayme Salsman and Elias Habib and {Van Iderstine}, Carter and Moamen Bydoun and Lewis, {Stephen M.} and Graham Dellaire",

note = "Funding Information: This work was funded by a Breast Cancer Society/QEII Foundation operating grant awarded to GD and an Operating Grant (#24135) from the Cancer Research Society and the Beatrice Hunter Cancer Research Institute (BHCRI) awarded to SML. GD and SML are Senior Scientists of the BHCRI, and both AC and LEC were supported by Cancer Research Training Program awards from the BHCRI with funds provided by CBCF—Atlantic, The Canadian Cancer Society, Nova Scotia Division as part of The Terry Fox Foundation Strategic Health Research Training (STIHR) Program in Cancer Research at the Canadian Institutes of Health Research (CIHR). SM is supported by a CGSM studentship from Natural Sciences and Engineering Research Council of Canada (NSERC), a Predoctoral Fellowship from the Killam Trusts, and a Scotia Scholar award from the Nova Scotia Health Research Foundation. Publisher Copyright: {\textcopyright} 2021 The Authors. The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology.",

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AU - Clarke, Livia E.

AU - Cook, Allyson

AU - Mathavarajah, Sabateeshan

AU - Bera, Amit

AU - Salsman, Jayme

AU - Habib, Elias

AU - Van Iderstine, Carter

AU - Bydoun, Moamen

AU - Lewis, Stephen M.

AU - Dellaire, Graham

N1 - Funding Information: This work was funded by a Breast Cancer Society/QEII Foundation operating grant awarded to GD and an Operating Grant (#24135) from the Cancer Research Society and the Beatrice Hunter Cancer Research Institute (BHCRI) awarded to SML. GD and SML are Senior Scientists of the BHCRI, and both AC and LEC were supported by Cancer Research Training Program awards from the BHCRI with funds provided by CBCF—Atlantic, The Canadian Cancer Society, Nova Scotia Division as part of The Terry Fox Foundation Strategic Health Research Training (STIHR) Program in Cancer Research at the Canadian Institutes of Health Research (CIHR). SM is supported by a CGSM studentship from Natural Sciences and Engineering Research Council of Canada (NSERC), a Predoctoral Fellowship from the Killam Trusts, and a Scotia Scholar award from the Nova Scotia Health Research Foundation. Publisher Copyright: © 2021 The Authors. The FASEB Journal published by Wiley Periodicals LLC on behalf of Federation of American Societies for Experimental Biology.

PY - 2021/11

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N2 - The pre-mRNA processing factor 4 kinase (PRP4K, also known as PRPF4B) is an essential gene. However, reduced PRP4K expression is associated with aggressive breast and ovarian cancer phenotypes including taxane therapy resistance, increased cell migration and invasion in vitro, and cancer metastasis in mice. These results are consistent with PRP4K being a haploinsufficient tumor suppressor. Increased cell migration and invasion is associated with epithelial-to-mesenchymal transition (EMT), but how reduced PRP4K levels affect normal epithelial cell migration or EMT has not been studied. Depletion of PRP4K by small hairpin RNA (shRNA) in non-transformed mammary epithelial cell lines (MCF10A, HMLE) reduced or had no effect on 2D migration in the scratch assay but resulted in greater invasive potential in 3D transwell assays. Depletion of PRP4K in mesenchymal triple-negative breast cancer cells (MDA-MB-231) resulted in both enhanced 2D migration and 3D invasion, with 3D invasion correlated with higher fibronectin levels in both MDA-MB-231 and MCF10A cells and without changes in E-cadherin. Induction of EMT in MCF10A cells, by treatment with WNT-5a and TGF-β1, or depletion of eukaryotic translation initiation factor 3e (eIF3e) by shRNA, resulted in significantly reduced PRP4K expression. Mechanistically, induction of EMT by WNT-5a/TGF-β1 reduced PRP4K transcript levels, whereas eIF3e depletion led to reduced PRP4K translation. Finally, reduced PRP4K levels after eIF3e depletion correlated with increased YAP activity and nuclear localization, both of which are reversed by overexpression of exogenous PRP4K. Thus, PRP4K is a haploinsufficient tumor suppressor negatively regulated by EMT, that when depleted in normal mammary cells can increase cell invasion without inducing full EMT.

AB - The pre-mRNA processing factor 4 kinase (PRP4K, also known as PRPF4B) is an essential gene. However, reduced PRP4K expression is associated with aggressive breast and ovarian cancer phenotypes including taxane therapy resistance, increased cell migration and invasion in vitro, and cancer metastasis in mice. These results are consistent with PRP4K being a haploinsufficient tumor suppressor. Increased cell migration and invasion is associated with epithelial-to-mesenchymal transition (EMT), but how reduced PRP4K levels affect normal epithelial cell migration or EMT has not been studied. Depletion of PRP4K by small hairpin RNA (shRNA) in non-transformed mammary epithelial cell lines (MCF10A, HMLE) reduced or had no effect on 2D migration in the scratch assay but resulted in greater invasive potential in 3D transwell assays. Depletion of PRP4K in mesenchymal triple-negative breast cancer cells (MDA-MB-231) resulted in both enhanced 2D migration and 3D invasion, with 3D invasion correlated with higher fibronectin levels in both MDA-MB-231 and MCF10A cells and without changes in E-cadherin. Induction of EMT in MCF10A cells, by treatment with WNT-5a and TGF-β1, or depletion of eukaryotic translation initiation factor 3e (eIF3e) by shRNA, resulted in significantly reduced PRP4K expression. Mechanistically, induction of EMT by WNT-5a/TGF-β1 reduced PRP4K transcript levels, whereas eIF3e depletion led to reduced PRP4K translation. Finally, reduced PRP4K levels after eIF3e depletion correlated with increased YAP activity and nuclear localization, both of which are reversed by overexpression of exogenous PRP4K. Thus, PRP4K is a haploinsufficient tumor suppressor negatively regulated by EMT, that when depleted in normal mammary cells can increase cell invasion without inducing full EMT.

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Haploinsufficient tumor suppressor PRP4K is negatively regulated during epithelial-to-mesenchymal transition

Abstract

Bibliographical note

ASJC Scopus Subject Areas

PubMed: MeSH publication types

UN SDGs

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