A nuclear cAMP microdomain suppresses tumor growth by Hippo pathway inactivation

Marek M. Drozdz; Ashley S. Doane; Rached Alkallas; Garrett Desman; Rohan Bareja; Michael Reilly; Jakyung Bang; Maftuna Yusupova; Jaewon You; Zuhal Eraslan; Jenny Z. Wang; Akanksha Verma; Kelsey Aguirre; Elsbeth Kane; Ian R. Watson; Olivier Elemento; Elena Piskounova; Taha Merghoub; Jonathan H. Zippin

doi:10.1016/j.celrep.2022.111412

A nuclear cAMP microdomain suppresses tumor growth by Hippo pathway inactivation

Marek M. Drozdz, Ashley S. Doane, Rached Alkallas, Garrett Desman, Rohan Bareja, Michael Reilly, Jakyung Bang, Maftuna Yusupova, Jaewon You, Zuhal Eraslan, Jenny Z. Wang, Akanksha Verma, Kelsey Aguirre, Elsbeth Kane, Ian R. Watson, Olivier Elemento, Elena Piskounova, Taha Merghoub, Jonathan H. Zippin

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

1 Scopus citations

Abstract

Cyclic AMP (cAMP) signaling is localized to multiple spatially distinct microdomains, but the role of cAMP microdomains in cancer cell biology is poorly understood. Here, we present a tunable genetic system that allows us to activate cAMP signaling in specific microdomains. We uncover a nuclear cAMP microdomain that activates a tumor-suppressive pathway in a broad range of cancers by inhibiting YAP, a key effector protein of the Hippo pathway, inside the nucleus. We show that nuclear cAMP induces a LATS-dependent pathway leading to phosphorylation of nuclear YAP solely at serine 397 and export of YAP from the nucleus with no change in YAP protein stability. Thus, nuclear cAMP inhibition of nuclear YAP is distinct from other known mechanisms of Hippo regulation. Pharmacologic targeting of specific cAMP microdomains remains an untapped therapeutic approach for cancer; thus, drugs directed at the nuclear cAMP microdomain may provide avenues for the treatment of cancer.

Original language	English
Article number	111412
Journal	Cell Reports
Volume	40
Issue number	13
DOIs	https://doi.org/10.1016/j.celrep.2022.111412
State	Published - 27 Sep 2022

Keywords

CP: Molecular biology
Hippo pathway
PKA
YAP
cAMP
nucleus
sAC
tumor suppression

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10.1016/j.celrep.2022.111412

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Drozdz, M. M., Doane, A. S., Alkallas, R., Desman, G., Bareja, R., Reilly, M., Bang, J., Yusupova, M., You, J., Eraslan, Z., Wang, J. Z., Verma, A., Aguirre, K., Kane, E., Watson, I. R., Elemento, O., Piskounova, E., Merghoub, T., & Zippin, J. H. (2022). A nuclear cAMP microdomain suppresses tumor growth by Hippo pathway inactivation. Cell Reports, 40(13), [111412]. https://doi.org/10.1016/j.celrep.2022.111412

@article{333d84c4622847d88034bb75fca5d2ce,

title = "A nuclear cAMP microdomain suppresses tumor growth by Hippo pathway inactivation",

abstract = "Cyclic AMP (cAMP) signaling is localized to multiple spatially distinct microdomains, but the role of cAMP microdomains in cancer cell biology is poorly understood. Here, we present a tunable genetic system that allows us to activate cAMP signaling in specific microdomains. We uncover a nuclear cAMP microdomain that activates a tumor-suppressive pathway in a broad range of cancers by inhibiting YAP, a key effector protein of the Hippo pathway, inside the nucleus. We show that nuclear cAMP induces a LATS-dependent pathway leading to phosphorylation of nuclear YAP solely at serine 397 and export of YAP from the nucleus with no change in YAP protein stability. Thus, nuclear cAMP inhibition of nuclear YAP is distinct from other known mechanisms of Hippo regulation. Pharmacologic targeting of specific cAMP microdomains remains an untapped therapeutic approach for cancer; thus, drugs directed at the nuclear cAMP microdomain may provide avenues for the treatment of cancer.",

keywords = "CP: Molecular biology, Hippo pathway, PKA, YAP, cAMP, nucleus, sAC, tumor suppression",

author = "Drozdz, {Marek M.} and Doane, {Ashley S.} and Rached Alkallas and Garrett Desman and Rohan Bareja and Michael Reilly and Jakyung Bang and Maftuna Yusupova and Jaewon You and Zuhal Eraslan and Wang, {Jenny Z.} and Akanksha Verma and Kelsey Aguirre and Elsbeth Kane and Watson, {Ian R.} and Olivier Elemento and Elena Piskounova and Taha Merghoub and Zippin, {Jonathan H.}",

note = "Funding Information: We thank members of the Zippin lab for critical reading of the manuscript and Paul Christos (Weill Cornell Medical College) for independent analysis of the statistics of the manuscript. We thank Dr. Jedd Wolchok for support for this work. We thank the following people for providing cell lines for this work: SCC12 were provided by Dr. Loraine Gudas (Weill Cornell Medical College, New York, NY, USA), colon cancer lines (DLD1 and SW480) were provided by Dr. Lukas Edward Dow (Weill Cornell Medical College, New York, NY, USA), pancreatic cancer lines (Hs766t, PANC1, MIAPaCa2) were provided by Dr. Lewis Cantley (Weill Cornell Medical College, New York, NY, USA), prostate cancer cell line LNCaP was provided by Dr. Christopher Barbieri (Weill Cornell Medical College, New York, NY, USA), and the melanoma M263 line was provided by Dr. Roger Lo (UCLA Medical Center). We thank Dr. Lukas Edward Dow for providing shRNA constructs against YAP and TAZ. We thank Peter T. Meinke of the Tri-Institutional Therapeutics Discovery Institute for providing TRULI inhibitor. We thank Kieran Harvey (Peter MacCallum Cancer Centre) for YAP 5SA MeWo RNA-seq datasets. J.H.Z. was funded in part by a Melanoma Research Alliance Team Science Award, Clinique Clinical Scholars Award, American Skin Association Calder Research Scholar Award, Pfizer ASPIRE award, NCI (1 R21 CA224391-01A1), and NIAMS (1 R01 AR077664-01A1). A.S.D. was funded in part by NCI (1 F31 CA220981-01). R.A. is a recipient of the CIHR Doctoral Award - Frederick Banting and Charles Best Canada Graduate Scholarships (CGS-D). O.E. was funded in part by R01 CA194547–04, U24CA210989-03, and P50 CA211024 CORE 2. T.M. was funded in part by the NIH/NCI R01 CA215136-01A1 and Cancer Center Support Grant P30 CA008748, the Swim Across America, Ludwig Institute for Cancer Research, Parker Institute for Cancer Immunotherapy, and Breast Cancer Research Foundation. E.P. was funded in part by R00CA201228-05, MRA Young Investigator Award, Emerson Collective Cancer Research Fund, and a Feldstein Medical Foundation Grant. This research was enabled in part by support provided by Calcul Qu{\'e}bec (https://www.calculquebec.ca/), SHARCNET (https://www.sharcnet.ca/), and Compute Canada (www.computecanada.ca), M.M.D. A.S.D. G.D. I.R.W. T.M. E.P. and J.H.Z. designed the experiments. M.M.D. A.S.D. G.D. R.B. M.R. J.B. M.Y. J.Y. Z.E. and R.A. generated the figures. M.M.D. J.Z.W. A.V. K.A. Z.E. and E.K. generated critical reagents or assisted with in vivo tumor experiments. M.M.D. O.E. T.M. E.P. I.R.W. and J.H.Z. wrote the manuscript with all authors providing feedback. J.H.Z. is a paid consultant and is on the medical advisory board of Hoth Therapeutics, is on the medical advisory board of SHADE, Inc. and is an inventor on US patent 8859213 on the use of antibodies directed against sACs for the diagnosis of melanocytic proliferations. O.E. is a cofounder and equity holder in Volastra Therapeutics and OneThree Biotech, an equity holder and SAB member in Owkin, Freenome, Genetic Intelligence, and Acuamark DX, and receives funding from Eli Lilly, Janssen, and Sanofi. T.M. is consultant for Leap Therapeutics, Immunos Therapeutics, and Pfizer, is a cofounder of Imvaq Therapeutics, has equity in Imvaq Therapeutics, reports grants from Bristol-Myers Squibb, Surface Oncology, Kyn Therapeutics, Infinity Pharmaceuticals, Peregrine Pharmeceuticals, Adaptive Biotechnologies, Leap Therapeutics, and Aprea, and is an inventor on patent applications related to work on oncolytic viral therapy, alphavirus-based vaccines, neo-antigen modeling, CD40, GITR, OX40, PD-1, and CTLA-4. Funding Information: We thank members of the Zippin lab for critical reading of the manuscript and Paul Christos (Weill Cornell Medical College) for independent analysis of the statistics of the manuscript. We thank Dr. Jedd Wolchok for support for this work. We thank the following people for providing cell lines for this work: SCC12 were provided by Dr. Loraine Gudas (Weill Cornell Medical College, New York, NY, USA), colon cancer lines (DLD1 and SW480) were provided by Dr. Lukas Edward Dow (Weill Cornell Medical College, New York, NY, USA), pancreatic cancer lines (Hs766t, PANC1, MIAPaCa2) were provided by Dr. Lewis Cantley (Weill Cornell Medical College, New York, NY, USA), prostate cancer cell line LNCaP was provided by Dr. Christopher Barbieri (Weill Cornell Medical College, New York, NY, USA), and the melanoma M263 line was provided by Dr. Roger Lo (UCLA Medical Center). We thank Dr. Lukas Edward Dow for providing shRNA constructs against YAP and TAZ. We thank Peter T. Meinke of the Tri-Institutional Therapeutics Discovery Institute for providing TRULI inhibitor. We thank Kieran Harvey (Peter MacCallum Cancer Centre) for YAP 5SA MeWo RNA-seq datasets. J.H.Z. was funded in part by a Melanoma Research Alliance Team Science Award, Clinique Clinical Scholars Award, American Skin Association Calder Research Scholar Award, Pfizer ASPIRE award, NCI ( 1 R21 CA224391-01A1 ), and NIAMS ( 1 R01 AR077664-01A1 ). A.S.D. was funded in part by NCI ( 1 F31 CA220981-01 ). R.A. is a recipient of the CIHR Doctoral Award - Frederick Banting and Charles Best Canada Graduate Scholarships (CGS-D). O.E. was funded in part by R01 CA194547–04 , U24CA210989-03 , and P50 CA211024 CORE 2. T.M. was funded in part by the NIH /NCI R01 CA215136-01A1 and Cancer Center Support Grant P30 CA008748 , the Swim Across America, Ludwig Institute for Cancer Research, Parker Institute for Cancer Immunotherapy, and Breast Cancer Research Foundation . E.P. was funded in part by R00CA201228-05 , MRA Young Investigator Award, Emerson Collective Cancer Research Fund, and a Feldstein Medical Foundation Grant. This research was enabled in part by support provided by Calcul Qu{\'e}bec ( https://www.calculquebec.ca/ ), SHARCNET ( https://www.sharcnet.ca/ ), and Compute Canada ( www.computecanada.ca ) Publisher Copyright: {\textcopyright} 2022 The Author(s)",

year = "2022",

month = sep,

day = "27",

doi = "10.1016/j.celrep.2022.111412",

language = "English",

volume = "40",

journal = "Cell Reports",

issn = "2211-1247",

publisher = "Cell Press",

number = "13",

}

Drozdz, MM, Doane, AS, Alkallas, R, Desman, G, Bareja, R, Reilly, M, Bang, J, Yusupova, M, You, J, Eraslan, Z, Wang, JZ, Verma, A, Aguirre, K, Kane, E, Watson, IR, Elemento, O, Piskounova, E, Merghoub, T & Zippin, JH 2022, 'A nuclear cAMP microdomain suppresses tumor growth by Hippo pathway inactivation', Cell Reports, vol. 40, no. 13, 111412. https://doi.org/10.1016/j.celrep.2022.111412

TY - JOUR

T1 - A nuclear cAMP microdomain suppresses tumor growth by Hippo pathway inactivation

AU - Drozdz, Marek M.

AU - Doane, Ashley S.

AU - Alkallas, Rached

AU - Desman, Garrett

AU - Bareja, Rohan

AU - Reilly, Michael

AU - Bang, Jakyung

AU - Yusupova, Maftuna

AU - You, Jaewon

AU - Eraslan, Zuhal

AU - Wang, Jenny Z.

AU - Verma, Akanksha

AU - Aguirre, Kelsey

AU - Kane, Elsbeth

AU - Watson, Ian R.

AU - Elemento, Olivier

AU - Piskounova, Elena

AU - Merghoub, Taha

AU - Zippin, Jonathan H.

N1 - Funding Information: We thank members of the Zippin lab for critical reading of the manuscript and Paul Christos (Weill Cornell Medical College) for independent analysis of the statistics of the manuscript. We thank Dr. Jedd Wolchok for support for this work. We thank the following people for providing cell lines for this work: SCC12 were provided by Dr. Loraine Gudas (Weill Cornell Medical College, New York, NY, USA), colon cancer lines (DLD1 and SW480) were provided by Dr. Lukas Edward Dow (Weill Cornell Medical College, New York, NY, USA), pancreatic cancer lines (Hs766t, PANC1, MIAPaCa2) were provided by Dr. Lewis Cantley (Weill Cornell Medical College, New York, NY, USA), prostate cancer cell line LNCaP was provided by Dr. Christopher Barbieri (Weill Cornell Medical College, New York, NY, USA), and the melanoma M263 line was provided by Dr. Roger Lo (UCLA Medical Center). We thank Dr. Lukas Edward Dow for providing shRNA constructs against YAP and TAZ. We thank Peter T. Meinke of the Tri-Institutional Therapeutics Discovery Institute for providing TRULI inhibitor. We thank Kieran Harvey (Peter MacCallum Cancer Centre) for YAP 5SA MeWo RNA-seq datasets. J.H.Z. was funded in part by a Melanoma Research Alliance Team Science Award, Clinique Clinical Scholars Award, American Skin Association Calder Research Scholar Award, Pfizer ASPIRE award, NCI (1 R21 CA224391-01A1), and NIAMS (1 R01 AR077664-01A1). A.S.D. was funded in part by NCI (1 F31 CA220981-01). R.A. is a recipient of the CIHR Doctoral Award - Frederick Banting and Charles Best Canada Graduate Scholarships (CGS-D). O.E. was funded in part by R01 CA194547–04, U24CA210989-03, and P50 CA211024 CORE 2. T.M. was funded in part by the NIH/NCI R01 CA215136-01A1 and Cancer Center Support Grant P30 CA008748, the Swim Across America, Ludwig Institute for Cancer Research, Parker Institute for Cancer Immunotherapy, and Breast Cancer Research Foundation. E.P. was funded in part by R00CA201228-05, MRA Young Investigator Award, Emerson Collective Cancer Research Fund, and a Feldstein Medical Foundation Grant. This research was enabled in part by support provided by Calcul Québec (https://www.calculquebec.ca/), SHARCNET (https://www.sharcnet.ca/), and Compute Canada (www.computecanada.ca), M.M.D. A.S.D. G.D. I.R.W. T.M. E.P. and J.H.Z. designed the experiments. M.M.D. A.S.D. G.D. R.B. M.R. J.B. M.Y. J.Y. Z.E. and R.A. generated the figures. M.M.D. J.Z.W. A.V. K.A. Z.E. and E.K. generated critical reagents or assisted with in vivo tumor experiments. M.M.D. O.E. T.M. E.P. I.R.W. and J.H.Z. wrote the manuscript with all authors providing feedback. J.H.Z. is a paid consultant and is on the medical advisory board of Hoth Therapeutics, is on the medical advisory board of SHADE, Inc. and is an inventor on US patent 8859213 on the use of antibodies directed against sACs for the diagnosis of melanocytic proliferations. O.E. is a cofounder and equity holder in Volastra Therapeutics and OneThree Biotech, an equity holder and SAB member in Owkin, Freenome, Genetic Intelligence, and Acuamark DX, and receives funding from Eli Lilly, Janssen, and Sanofi. T.M. is consultant for Leap Therapeutics, Immunos Therapeutics, and Pfizer, is a cofounder of Imvaq Therapeutics, has equity in Imvaq Therapeutics, reports grants from Bristol-Myers Squibb, Surface Oncology, Kyn Therapeutics, Infinity Pharmaceuticals, Peregrine Pharmeceuticals, Adaptive Biotechnologies, Leap Therapeutics, and Aprea, and is an inventor on patent applications related to work on oncolytic viral therapy, alphavirus-based vaccines, neo-antigen modeling, CD40, GITR, OX40, PD-1, and CTLA-4. Funding Information: We thank members of the Zippin lab for critical reading of the manuscript and Paul Christos (Weill Cornell Medical College) for independent analysis of the statistics of the manuscript. We thank Dr. Jedd Wolchok for support for this work. We thank the following people for providing cell lines for this work: SCC12 were provided by Dr. Loraine Gudas (Weill Cornell Medical College, New York, NY, USA), colon cancer lines (DLD1 and SW480) were provided by Dr. Lukas Edward Dow (Weill Cornell Medical College, New York, NY, USA), pancreatic cancer lines (Hs766t, PANC1, MIAPaCa2) were provided by Dr. Lewis Cantley (Weill Cornell Medical College, New York, NY, USA), prostate cancer cell line LNCaP was provided by Dr. Christopher Barbieri (Weill Cornell Medical College, New York, NY, USA), and the melanoma M263 line was provided by Dr. Roger Lo (UCLA Medical Center). We thank Dr. Lukas Edward Dow for providing shRNA constructs against YAP and TAZ. We thank Peter T. Meinke of the Tri-Institutional Therapeutics Discovery Institute for providing TRULI inhibitor. We thank Kieran Harvey (Peter MacCallum Cancer Centre) for YAP 5SA MeWo RNA-seq datasets. J.H.Z. was funded in part by a Melanoma Research Alliance Team Science Award, Clinique Clinical Scholars Award, American Skin Association Calder Research Scholar Award, Pfizer ASPIRE award, NCI ( 1 R21 CA224391-01A1 ), and NIAMS ( 1 R01 AR077664-01A1 ). A.S.D. was funded in part by NCI ( 1 F31 CA220981-01 ). R.A. is a recipient of the CIHR Doctoral Award - Frederick Banting and Charles Best Canada Graduate Scholarships (CGS-D). O.E. was funded in part by R01 CA194547–04 , U24CA210989-03 , and P50 CA211024 CORE 2. T.M. was funded in part by the NIH /NCI R01 CA215136-01A1 and Cancer Center Support Grant P30 CA008748 , the Swim Across America, Ludwig Institute for Cancer Research, Parker Institute for Cancer Immunotherapy, and Breast Cancer Research Foundation . E.P. was funded in part by R00CA201228-05 , MRA Young Investigator Award, Emerson Collective Cancer Research Fund, and a Feldstein Medical Foundation Grant. This research was enabled in part by support provided by Calcul Québec ( https://www.calculquebec.ca/ ), SHARCNET ( https://www.sharcnet.ca/ ), and Compute Canada ( www.computecanada.ca ) Publisher Copyright: © 2022 The Author(s)

PY - 2022/9/27

Y1 - 2022/9/27

N2 - Cyclic AMP (cAMP) signaling is localized to multiple spatially distinct microdomains, but the role of cAMP microdomains in cancer cell biology is poorly understood. Here, we present a tunable genetic system that allows us to activate cAMP signaling in specific microdomains. We uncover a nuclear cAMP microdomain that activates a tumor-suppressive pathway in a broad range of cancers by inhibiting YAP, a key effector protein of the Hippo pathway, inside the nucleus. We show that nuclear cAMP induces a LATS-dependent pathway leading to phosphorylation of nuclear YAP solely at serine 397 and export of YAP from the nucleus with no change in YAP protein stability. Thus, nuclear cAMP inhibition of nuclear YAP is distinct from other known mechanisms of Hippo regulation. Pharmacologic targeting of specific cAMP microdomains remains an untapped therapeutic approach for cancer; thus, drugs directed at the nuclear cAMP microdomain may provide avenues for the treatment of cancer.

AB - Cyclic AMP (cAMP) signaling is localized to multiple spatially distinct microdomains, but the role of cAMP microdomains in cancer cell biology is poorly understood. Here, we present a tunable genetic system that allows us to activate cAMP signaling in specific microdomains. We uncover a nuclear cAMP microdomain that activates a tumor-suppressive pathway in a broad range of cancers by inhibiting YAP, a key effector protein of the Hippo pathway, inside the nucleus. We show that nuclear cAMP induces a LATS-dependent pathway leading to phosphorylation of nuclear YAP solely at serine 397 and export of YAP from the nucleus with no change in YAP protein stability. Thus, nuclear cAMP inhibition of nuclear YAP is distinct from other known mechanisms of Hippo regulation. Pharmacologic targeting of specific cAMP microdomains remains an untapped therapeutic approach for cancer; thus, drugs directed at the nuclear cAMP microdomain may provide avenues for the treatment of cancer.

KW - CP: Molecular biology

KW - Hippo pathway

KW - PKA

KW - YAP

KW - cAMP

KW - nucleus

KW - sAC

KW - tumor suppression

UR - http://www.scopus.com/inward/record.url?scp=85138779595&partnerID=8YFLogxK

U2 - 10.1016/j.celrep.2022.111412

DO - 10.1016/j.celrep.2022.111412

M3 - Article

C2 - 36170819

AN - SCOPUS:85138779595

SN - 2211-1247

VL - 40

JO - Cell Reports

JF - Cell Reports

IS - 13

M1 - 111412

ER -

A nuclear cAMP microdomain suppresses tumor growth by Hippo pathway inactivation

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