Leukemogenic nucleophosmin mutation disrupts the transcription factor hub that regulates granulomonocytic fates

Xiaorong Gu; Quteba Ebrahem; Reda Z. Mahfouz; Metis Hasipek; Francis Enane; Tomas Radivoyevitch; Nicolas Rapin; Bartlomiej Przychodzen; Zhenbo Hu; Ramesh Balusu; Claudiu V. Cotta; David Wald; Christian Argueta; Yosef Landesman; Maria Paola Martelli; Brunangelo Falini; Hetty Carraway; Bo T. Porse; Jaroslaw Maciejewski; Babal K. Jha; Yogen Saunthararajah

doi:10.1172/jci97117

Leukemogenic nucleophosmin mutation disrupts the transcription factor hub that regulates granulomonocytic fates

Xiaorong Gu, Quteba Ebrahem, Reda Z. Mahfouz, Metis Hasipek, Francis Enane, Tomas Radivoyevitch, Nicolas Rapin, Bartlomiej Przychodzen, Zhenbo Hu, Ramesh Balusu, Claudiu V. Cotta, David Wald, Christian Argueta, Yosef Landesman, Maria Paola Martelli, Brunangelo Falini, Hetty Carraway, Bo T. Porse, Jaroslaw Maciejewski, Babal K. JhaYogen Saunthararajah

48 Citations (Scopus)

Abstract

Nucleophosmin (NPM1) is among the most frequently mutated genes in acute myeloid leukemia (AML). It is not known, however, how the resulting oncoprotein mutant NPM1 is leukemogenic. To reveal the cellular machinery in which NPM1 participates in myeloid cells, we analyzed the endogenous NPM1 protein interactome by mass spectrometry and discovered abundant amounts of the master transcription factor driver of monocyte lineage differentiation PU.1 (also known as SPI1). Mutant NPM1, which aberrantly accumulates in cytoplasm, dislocated PU.1 into cytoplasm with it. CEBPA and RUNX1, the master transcription factors that collaborate with PU.1 to activate granulomonocytic lineage fates, remained nuclear; but without PU.1, their coregulator interactions were toggled from coactivators to corepressors, repressing instead of activating more than 500 granulocyte and monocyte terminal differentiation genes. An inhibitor of nuclear export, selinexor, by locking mutant NPM1/PU.1 in the nucleus, activated terminal monocytic fates. Direct depletion of the corepressor DNA methyltransferase 1 (DNMT1) from the CEBPA/RUNX1 protein interactome using the clinical drug decitabine activated terminal granulocytic fates. Together, these noncytotoxic treatments extended survival by more than 160 days versus vehicle in a patient-derived xenotransplant model of NPM1/FLT3-mutated AML. In sum, mutant NPM1 represses monocyte and granulocyte terminal differentiation by disrupting PU.1/CEBPA/RUNX1 collaboration, a transforming action that can be reversed by pharmacodynamically directed dosing of clinical small molecules.

Original language	English
Journal	The Journal of Clinical Investigation
Volume	128
Issue number	10
Pages (from-to)	4260-4279
Number of pages	20
ISSN	1558-8238
DOIs	https://doi.org/10.1172/jci97117
Publication status	Published - 1 Oct 2018

Keywords

epigenetics
hematology
leukemias
oncology
transport

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Gu, X., Ebrahem, Q., Mahfouz, R. Z., Hasipek, M., Enane, F., Radivoyevitch, T., Rapin, N., Przychodzen, B., Hu, Z., Balusu, R., Cotta, C. V., Wald, D., Argueta, C., Landesman, Y., Martelli, M. P., Falini, B., Carraway, H., Porse, B. T., Maciejewski, J., ... Saunthararajah, Y. (2018). Leukemogenic nucleophosmin mutation disrupts the transcription factor hub that regulates granulomonocytic fates. The Journal of Clinical Investigation, 128(10), 4260-4279. https://doi.org/10.1172/jci97117

Gu, X, Ebrahem, Q, Mahfouz, RZ, Hasipek, M, Enane, F, Radivoyevitch, T, Rapin, N, Przychodzen, B, Hu, Z, Balusu, R, Cotta, CV, Wald, D, Argueta, C, Landesman, Y, Martelli, MP, Falini, B, Carraway, H, Porse, BT, Maciejewski, J, Jha, BK & Saunthararajah, Y 2018, 'Leukemogenic nucleophosmin mutation disrupts the transcription factor hub that regulates granulomonocytic fates', The Journal of Clinical Investigation, vol. 128, no. 10, pp. 4260-4279. https://doi.org/10.1172/jci97117

@article{38abd0dcfb7c44e880665f605dbe0119,

title = "Leukemogenic nucleophosmin mutation disrupts the transcription factor hub that regulates granulomonocytic fates",

abstract = "Nucleophosmin (NPM1) is among the most frequently mutated genes in acute myeloid leukemia (AML). It is not known, however, how the resulting oncoprotein mutant NPM1 is leukemogenic. To reveal the cellular machinery in which NPM1 participates in myeloid cells, we analyzed the endogenous NPM1 protein interactome by mass spectrometry and discovered abundant amounts of the master transcription factor driver of monocyte lineage differentiation PU.1 (also known as SPI1). Mutant NPM1, which aberrantly accumulates in cytoplasm, dislocated PU.1 into cytoplasm with it. CEBPA and RUNX1, the master transcription factors that collaborate with PU.1 to activate granulomonocytic lineage fates, remained nuclear; but without PU.1, their coregulator interactions were toggled from coactivators to corepressors, repressing instead of activating more than 500 granulocyte and monocyte terminal differentiation genes. An inhibitor of nuclear export, selinexor, by locking mutant NPM1/PU.1 in the nucleus, activated terminal monocytic fates. Direct depletion of the corepressor DNA methyltransferase 1 (DNMT1) from the CEBPA/RUNX1 protein interactome using the clinical drug decitabine activated terminal granulocytic fates. Together, these noncytotoxic treatments extended survival by more than 160 days versus vehicle in a patient-derived xenotransplant model of NPM1/FLT3-mutated AML. In sum, mutant NPM1 represses monocyte and granulocyte terminal differentiation by disrupting PU.1/CEBPA/RUNX1 collaboration, a transforming action that can be reversed by pharmacodynamically directed dosing of clinical small molecules.",

keywords = "epigenetics, hematology, leukemias, oncology, transport",

author = "Xiaorong Gu and Quteba Ebrahem and Mahfouz, {Reda Z.} and Metis Hasipek and Francis Enane and Tomas Radivoyevitch and Nicolas Rapin and Bartlomiej Przychodzen and Zhenbo Hu and Ramesh Balusu and Cotta, {Claudiu V.} and David Wald and Christian Argueta and Yosef Landesman and Martelli, {Maria Paola} and Brunangelo Falini and Hetty Carraway and Porse, {Bo T.} and Jaroslaw Maciejewski and Jha, {Babal K.} and Yogen Saunthararajah",

year = "2018",

month = oct,

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doi = "10.1172/jci97117",

language = "English",

volume = "128",

pages = "4260--4279",

journal = "The Journal of Clinical Investigation",

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publisher = "American Society for Clinical Investigation",

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TY - JOUR

T1 - Leukemogenic nucleophosmin mutation disrupts the transcription factor hub that regulates granulomonocytic fates

AU - Gu, Xiaorong

AU - Ebrahem, Quteba

AU - Mahfouz, Reda Z.

AU - Hasipek, Metis

AU - Enane, Francis

AU - Radivoyevitch, Tomas

AU - Rapin, Nicolas

AU - Przychodzen, Bartlomiej

AU - Hu, Zhenbo

AU - Balusu, Ramesh

AU - Cotta, Claudiu V.

AU - Wald, David

AU - Argueta, Christian

AU - Landesman, Yosef

AU - Martelli, Maria Paola

AU - Falini, Brunangelo

AU - Carraway, Hetty

AU - Porse, Bo T.

AU - Maciejewski, Jaroslaw

AU - Jha, Babal K.

AU - Saunthararajah, Yogen

PY - 2018/10/1

Y1 - 2018/10/1

N2 - Nucleophosmin (NPM1) is among the most frequently mutated genes in acute myeloid leukemia (AML). It is not known, however, how the resulting oncoprotein mutant NPM1 is leukemogenic. To reveal the cellular machinery in which NPM1 participates in myeloid cells, we analyzed the endogenous NPM1 protein interactome by mass spectrometry and discovered abundant amounts of the master transcription factor driver of monocyte lineage differentiation PU.1 (also known as SPI1). Mutant NPM1, which aberrantly accumulates in cytoplasm, dislocated PU.1 into cytoplasm with it. CEBPA and RUNX1, the master transcription factors that collaborate with PU.1 to activate granulomonocytic lineage fates, remained nuclear; but without PU.1, their coregulator interactions were toggled from coactivators to corepressors, repressing instead of activating more than 500 granulocyte and monocyte terminal differentiation genes. An inhibitor of nuclear export, selinexor, by locking mutant NPM1/PU.1 in the nucleus, activated terminal monocytic fates. Direct depletion of the corepressor DNA methyltransferase 1 (DNMT1) from the CEBPA/RUNX1 protein interactome using the clinical drug decitabine activated terminal granulocytic fates. Together, these noncytotoxic treatments extended survival by more than 160 days versus vehicle in a patient-derived xenotransplant model of NPM1/FLT3-mutated AML. In sum, mutant NPM1 represses monocyte and granulocyte terminal differentiation by disrupting PU.1/CEBPA/RUNX1 collaboration, a transforming action that can be reversed by pharmacodynamically directed dosing of clinical small molecules.

AB - Nucleophosmin (NPM1) is among the most frequently mutated genes in acute myeloid leukemia (AML). It is not known, however, how the resulting oncoprotein mutant NPM1 is leukemogenic. To reveal the cellular machinery in which NPM1 participates in myeloid cells, we analyzed the endogenous NPM1 protein interactome by mass spectrometry and discovered abundant amounts of the master transcription factor driver of monocyte lineage differentiation PU.1 (also known as SPI1). Mutant NPM1, which aberrantly accumulates in cytoplasm, dislocated PU.1 into cytoplasm with it. CEBPA and RUNX1, the master transcription factors that collaborate with PU.1 to activate granulomonocytic lineage fates, remained nuclear; but without PU.1, their coregulator interactions were toggled from coactivators to corepressors, repressing instead of activating more than 500 granulocyte and monocyte terminal differentiation genes. An inhibitor of nuclear export, selinexor, by locking mutant NPM1/PU.1 in the nucleus, activated terminal monocytic fates. Direct depletion of the corepressor DNA methyltransferase 1 (DNMT1) from the CEBPA/RUNX1 protein interactome using the clinical drug decitabine activated terminal granulocytic fates. Together, these noncytotoxic treatments extended survival by more than 160 days versus vehicle in a patient-derived xenotransplant model of NPM1/FLT3-mutated AML. In sum, mutant NPM1 represses monocyte and granulocyte terminal differentiation by disrupting PU.1/CEBPA/RUNX1 collaboration, a transforming action that can be reversed by pharmacodynamically directed dosing of clinical small molecules.

KW - epigenetics

KW - hematology

KW - leukemias

KW - oncology

KW - transport

U2 - 10.1172/jci97117

DO - 10.1172/jci97117

M3 - Journal article

C2 - 30015632

SN - 1558-8238

VL - 128

SP - 4260

EP - 4279

JO - The Journal of Clinical Investigation

JF - The Journal of Clinical Investigation

IS - 10

ER -

Leukemogenic nucleophosmin mutation disrupts the transcription factor hub that regulates granulomonocytic fates

Abstract

Keywords

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