Effects of novel pyrrolomycin MP1 in MYCN amplified chemoresistant neuroblastoma cell lines alone and combined with temsirolimus

Timothy R McGuire; Don W. Coulter; Dachang Bai; Jason A. Sughroue; Jerry Li; Zunhua Yang; Zhen Qiao; Yan Liu; Daryl J. Murry; Yashpal S. Chhonker; Erin M. McIntyre; Gracey Alexander; John G Sharp; Rongshi Li

doi:10.1186/s12885-019-6033-2

Effects of novel pyrrolomycin MP1 in MYCN amplified chemoresistant neuroblastoma cell lines alone and combined with temsirolimus

Timothy R McGuire, Don W. Coulter, Dachang Bai, Jason A. Sughroue, Jerry Li, Zunhua Yang, Zhen Qiao, Yan Liu, Daryl J. Murry, Yashpal S. Chhonker, Erin M. McIntyre, Gracey Alexander, John G Sharp, Rongshi Li

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Abstract

Background: The activity of MP1, a pyrrolomycin, was studied in MYCN amplified neuroblastoma (NB) alone and combined with temsirolimus (TEM). Methods: Activity of MP1 was tested in MYCN amplified (BE-2c, IMR) and non amplified (SKN-AS) NB cells. The effect of MP1 on MYCN, MCL-1, cleaved PARP, LC3II/LC3I, bcl-2, BAX, and BRD-4 were determined by western blot and RNAseq. The effect of MP1 on metabolism, mitochondrial morphology, and cell cycle was determined. Toxicology and efficacy of MP1 plus TEM were evaluated. Results: The IC₅₀ of MP1 was 0.096 μM in BE-2c cells compared to 0.89 μM in IMR, and >50 μM in SKN-AS. The IC₅₀ of MP1 plus TEM in BE-2c cells was 0.023 μM. MP1 inhibited metabolism leading to quiescence and produced a decline in cell cycle S-phase. Electron microscopy showed cristae loss and rounding up of mitochondria. Gene and protein expression for MYCN and MCL-1 declined while LCII and cleaved PARP increased. Protein expression of BAX, bcl-2, and BRD-4 were not significantly changed after MP1 treatment. The in-vivo concentrations of MP1 in blood and tumor were sufficient to produce the biologic effects seen in-vitro. MP1 plus TEM produced a complete response in 3 out of 5 tumor bearing mice. In a second mouse study, the combination of MP1 and TEM slowed tumor growth compared to control. Conclusions: MP1 has a potent inhibitory effect on the viability of MYCN amplified NB. Inhibition of metabolism by MP1 induced quiescence and autophagy with a favorable toxicology and drug distribution profile. When combined with TEM anti-tumor activity was potentiated in-vitro and in-vivo.

Original language	English (US)
Article number	837
Journal	BMC cancer
Volume	19
Issue number	1
DOIs	https://doi.org/10.1186/s12885-019-6033-2
State	Published - Aug 27 2019

Keywords

MYCN
Metabolism
Mitochondria
Neuroblastoma
Pyrrolomycin Marinopyrrole (MP1)
Temsirolimus

ASJC Scopus subject areas

Oncology
Genetics
Cancer Research

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10.1186/s12885-019-6033-2

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McGuire, T. R., Coulter, D. W., Bai, D., Sughroue, J. A., Li, J., Yang, Z., Qiao, Z., Liu, Y., Murry, D. J., Chhonker, Y. S., McIntyre, E. M., Alexander, G., Sharp, J. G., & Li, R. (2019). Effects of novel pyrrolomycin MP1 in MYCN amplified chemoresistant neuroblastoma cell lines alone and combined with temsirolimus. BMC cancer, 19(1), Article 837. https://doi.org/10.1186/s12885-019-6033-2

@article{743162ac9bce4358983417893efefa11,

title = "Effects of novel pyrrolomycin MP1 in MYCN amplified chemoresistant neuroblastoma cell lines alone and combined with temsirolimus",

abstract = "Background: The activity of MP1, a pyrrolomycin, was studied in MYCN amplified neuroblastoma (NB) alone and combined with temsirolimus (TEM). Methods: Activity of MP1 was tested in MYCN amplified (BE-2c, IMR) and non amplified (SKN-AS) NB cells. The effect of MP1 on MYCN, MCL-1, cleaved PARP, LC3II/LC3I, bcl-2, BAX, and BRD-4 were determined by western blot and RNAseq. The effect of MP1 on metabolism, mitochondrial morphology, and cell cycle was determined. Toxicology and efficacy of MP1 plus TEM were evaluated. Results: The IC50 of MP1 was 0.096 μM in BE-2c cells compared to 0.89 μM in IMR, and >50 μM in SKN-AS. The IC50 of MP1 plus TEM in BE-2c cells was 0.023 μM. MP1 inhibited metabolism leading to quiescence and produced a decline in cell cycle S-phase. Electron microscopy showed cristae loss and rounding up of mitochondria. Gene and protein expression for MYCN and MCL-1 declined while LCII and cleaved PARP increased. Protein expression of BAX, bcl-2, and BRD-4 were not significantly changed after MP1 treatment. The in-vivo concentrations of MP1 in blood and tumor were sufficient to produce the biologic effects seen in-vitro. MP1 plus TEM produced a complete response in 3 out of 5 tumor bearing mice. In a second mouse study, the combination of MP1 and TEM slowed tumor growth compared to control. Conclusions: MP1 has a potent inhibitory effect on the viability of MYCN amplified NB. Inhibition of metabolism by MP1 induced quiescence and autophagy with a favorable toxicology and drug distribution profile. When combined with TEM anti-tumor activity was potentiated in-vitro and in-vivo.",

keywords = "MYCN, Metabolism, Mitochondria, Neuroblastoma, Pyrrolomycin Marinopyrrole (MP1), Temsirolimus",

author = "McGuire, {Timothy R} and Coulter, {Don W.} and Dachang Bai and Sughroue, {Jason A.} and Jerry Li and Zunhua Yang and Zhen Qiao and Yan Liu and Murry, {Daryl J.} and Chhonker, {Yashpal S.} and McIntyre, {Erin M.} and Gracey Alexander and Sharp, {John G} and Rongshi Li",

note = "Funding Information: The authors thank the Flow Cytometry Core facilities at Creighton University Medical Center, the UNMC EM Core, and UNMC Molecular Biology Core for their help in these studies. We thank the State of Nebraska and UNMC for Nebraska Research Initiative Grant and Fred and Pamela Buffett Cancer Center Support Grant from the National Cancer Institute (P30 CA036727). Funding Information: The authors thank the support of the State of Nebraska through the Pediatric Cancer Research Grant (LB905) awarded to D.W. Coulter, MD. The funding agency had no role in the design of the study and collection, analysis, and interpretation of data and in writing the manuscript. Publisher Copyright: {\textcopyright} 2019 The Author(s).",

year = "2019",

month = aug,

day = "27",

doi = "10.1186/s12885-019-6033-2",

language = "English (US)",

volume = "19",

journal = "BMC cancer",

issn = "1471-2407",

publisher = "BioMed Central",

number = "1",

}

TY - JOUR

T1 - Effects of novel pyrrolomycin MP1 in MYCN amplified chemoresistant neuroblastoma cell lines alone and combined with temsirolimus

AU - McGuire, Timothy R

AU - Coulter, Don W.

AU - Bai, Dachang

AU - Sughroue, Jason A.

AU - Li, Jerry

AU - Yang, Zunhua

AU - Qiao, Zhen

AU - Liu, Yan

AU - Murry, Daryl J.

AU - Chhonker, Yashpal S.

AU - McIntyre, Erin M.

AU - Alexander, Gracey

AU - Sharp, John G

AU - Li, Rongshi

N1 - Funding Information: The authors thank the Flow Cytometry Core facilities at Creighton University Medical Center, the UNMC EM Core, and UNMC Molecular Biology Core for their help in these studies. We thank the State of Nebraska and UNMC for Nebraska Research Initiative Grant and Fred and Pamela Buffett Cancer Center Support Grant from the National Cancer Institute (P30 CA036727). Funding Information: The authors thank the support of the State of Nebraska through the Pediatric Cancer Research Grant (LB905) awarded to D.W. Coulter, MD. The funding agency had no role in the design of the study and collection, analysis, and interpretation of data and in writing the manuscript. Publisher Copyright: © 2019 The Author(s).

PY - 2019/8/27

Y1 - 2019/8/27

N2 - Background: The activity of MP1, a pyrrolomycin, was studied in MYCN amplified neuroblastoma (NB) alone and combined with temsirolimus (TEM). Methods: Activity of MP1 was tested in MYCN amplified (BE-2c, IMR) and non amplified (SKN-AS) NB cells. The effect of MP1 on MYCN, MCL-1, cleaved PARP, LC3II/LC3I, bcl-2, BAX, and BRD-4 were determined by western blot and RNAseq. The effect of MP1 on metabolism, mitochondrial morphology, and cell cycle was determined. Toxicology and efficacy of MP1 plus TEM were evaluated. Results: The IC50 of MP1 was 0.096 μM in BE-2c cells compared to 0.89 μM in IMR, and >50 μM in SKN-AS. The IC50 of MP1 plus TEM in BE-2c cells was 0.023 μM. MP1 inhibited metabolism leading to quiescence and produced a decline in cell cycle S-phase. Electron microscopy showed cristae loss and rounding up of mitochondria. Gene and protein expression for MYCN and MCL-1 declined while LCII and cleaved PARP increased. Protein expression of BAX, bcl-2, and BRD-4 were not significantly changed after MP1 treatment. The in-vivo concentrations of MP1 in blood and tumor were sufficient to produce the biologic effects seen in-vitro. MP1 plus TEM produced a complete response in 3 out of 5 tumor bearing mice. In a second mouse study, the combination of MP1 and TEM slowed tumor growth compared to control. Conclusions: MP1 has a potent inhibitory effect on the viability of MYCN amplified NB. Inhibition of metabolism by MP1 induced quiescence and autophagy with a favorable toxicology and drug distribution profile. When combined with TEM anti-tumor activity was potentiated in-vitro and in-vivo.

AB - Background: The activity of MP1, a pyrrolomycin, was studied in MYCN amplified neuroblastoma (NB) alone and combined with temsirolimus (TEM). Methods: Activity of MP1 was tested in MYCN amplified (BE-2c, IMR) and non amplified (SKN-AS) NB cells. The effect of MP1 on MYCN, MCL-1, cleaved PARP, LC3II/LC3I, bcl-2, BAX, and BRD-4 were determined by western blot and RNAseq. The effect of MP1 on metabolism, mitochondrial morphology, and cell cycle was determined. Toxicology and efficacy of MP1 plus TEM were evaluated. Results: The IC50 of MP1 was 0.096 μM in BE-2c cells compared to 0.89 μM in IMR, and >50 μM in SKN-AS. The IC50 of MP1 plus TEM in BE-2c cells was 0.023 μM. MP1 inhibited metabolism leading to quiescence and produced a decline in cell cycle S-phase. Electron microscopy showed cristae loss and rounding up of mitochondria. Gene and protein expression for MYCN and MCL-1 declined while LCII and cleaved PARP increased. Protein expression of BAX, bcl-2, and BRD-4 were not significantly changed after MP1 treatment. The in-vivo concentrations of MP1 in blood and tumor were sufficient to produce the biologic effects seen in-vitro. MP1 plus TEM produced a complete response in 3 out of 5 tumor bearing mice. In a second mouse study, the combination of MP1 and TEM slowed tumor growth compared to control. Conclusions: MP1 has a potent inhibitory effect on the viability of MYCN amplified NB. Inhibition of metabolism by MP1 induced quiescence and autophagy with a favorable toxicology and drug distribution profile. When combined with TEM anti-tumor activity was potentiated in-vitro and in-vivo.

KW - MYCN

KW - Metabolism

KW - Mitochondria

KW - Neuroblastoma

KW - Pyrrolomycin Marinopyrrole (MP1)

KW - Temsirolimus

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U2 - 10.1186/s12885-019-6033-2

DO - 10.1186/s12885-019-6033-2

M3 - Article

C2 - 31455317

AN - SCOPUS:85071638053

SN - 1471-2407

VL - 19

JO - BMC cancer

JF - BMC cancer

IS - 1

M1 - 837

ER -

Effects of novel pyrrolomycin MP1 in MYCN amplified chemoresistant neuroblastoma cell lines alone and combined with temsirolimus

Abstract

Keywords

ASJC Scopus subject areas

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