Inhibition of RNA Binding in SND1 Increases the Levels of miR-1-3p and Sensitizes Cancer Cells to Navitoclax

Saara Lehmusvaara; Teemu Haikarainen; Juha Saarikettu; Guillermo Martinez Nieto; Olli Silvennoinen

doi:10.3390/cancers14133100

Inhibition of RNA Binding in SND1 Increases the Levels of miR-1-3p and Sensitizes Cancer Cells to Navitoclax

Saara Lehmusvaara
, Teemu Haikarainen
, Juha Saarikettu
, Guillermo Martinez Nieto
, Olli Silvennoinen^*

^*Tämän työn vastaava kirjoittaja

Tutkimustuotos: Artikkeli › Tieteellinen › vertaisarvioitu

10 Sitaatiot (Scopus)

36 Lataukset (Pure)

Abstrakti

SND1 is an RNA-binding protein overexpressed in large variety of cancers. SND1 has been proposed to enhance stress tolerance in cancer cells, but the molecular mechanisms are still poorly understood. We analyzed the expression of 372 miRNAs in the colon carcinoma cell line and show that SND1 silencing increases the expression levels of several tumor suppressor miRNAs. Furthermore, SND1 knockdown showed synergetic effects with cancer drugs through MEK-ERK and Bcl-2 family-related apoptotic pathways. To explore whether the SND1-mediated RNA bind-ing/degradation is responsible for the observed effect, we developed a screening assay to identify small molecules that inhibit the RNA-binding function of SND1. The screen identified P2X purinoreceptor antagonists as the most potent inhibitors. Validation confirmed that the best hit, suramin, inhibits the RNA binding ability of SND1. The binding characteristics and mode of suramin to SND1 were characterized biophysically and by molecular docking that identified positively charged binding cavities in Staphylococcus nuclease domains. Importantly, suramin-mediated inhibition of RNA binding increased the expression of miR-1-3p, and enhanced sensitivity of cancer cells to Bcl-2 inhibitor navitoclax treatment. Taken together, we demonstrate as proof-of-concept a mechanism and an inhibitor compound for SND1 regulation of the survival of cancer cells through tumor suppressor miRNAs.

Alkuperäiskieli	Englanti
Artikkeli	3100
Sivumäärä	17
Julkaisu	Cancers
Vuosikerta	14
Numero	13
DOI - pysyväislinkit	https://doi.org/10.3390/cancers14133100
Tila	Julkaistu - kesäk. 2022
OKM-julkaisutyyppi	A1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä

Rahoitus

Acknowledgments: The authors want to thank Antti Kurttila, Sanna Liukkonen and Merja Peltomaa for their valuable technical help. The authors also thank the Tampere facility of Virus Production for their service, and the DDCB core facility supported by the University of Helsinki and Biocenter Finland. The authors also thank Laura Turunen at the High Throughput Biomedicine Unit at the Institute for Molecular Medicine Finland (FIMM). Funding: This research was funded by the Academy of Finland (grant number 335437), Sigrid Jusélius Foundation (grant number 1152), Finnish Cancer Foundation (grant number 4708167), Jane and Aatos Erkko Foundation, Tampere Tuberculosis Foundation (grant number 2019), and Pirkanmaa hospital district competitive research funding.

YK:n kestävän kehityksen tavoitteet

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SDG 3 – Hyvä terveys ja hyvinvointi

Julkaisufoorumi-taso

Jufo-taso 1

!!ASJC Scopus subject areas

Oncology
Cancer Research

Pääsy asiakirjaan

10.3390/cancers14133100Lisenssi: CC BY

cancers-14-03100Final published version, 2,11 MBLisenssi: CC BY

https://urn.fi/URN:NBN:fi:tuni-202208036201Lisenssi: CC BY

Siteeraa tätä

@article{adf3bb8f0bc44bb1acdd9168df6bb018,

title = "Inhibition of RNA Binding in SND1 Increases the Levels of miR-1-3p and Sensitizes Cancer Cells to Navitoclax",

abstract = "SND1 is an RNA-binding protein overexpressed in large variety of cancers. SND1 has been proposed to enhance stress tolerance in cancer cells, but the molecular mechanisms are still poorly understood. We analyzed the expression of 372 miRNAs in the colon carcinoma cell line and show that SND1 silencing increases the expression levels of several tumor suppressor miRNAs. Furthermore, SND1 knockdown showed synergetic effects with cancer drugs through MEK-ERK and Bcl-2 family-related apoptotic pathways. To explore whether the SND1-mediated RNA bind-ing/degradation is responsible for the observed effect, we developed a screening assay to identify small molecules that inhibit the RNA-binding function of SND1. The screen identified P2X purinoreceptor antagonists as the most potent inhibitors. Validation confirmed that the best hit, suramin, inhibits the RNA binding ability of SND1. The binding characteristics and mode of suramin to SND1 were characterized biophysically and by molecular docking that identified positively charged binding cavities in Staphylococcus nuclease domains. Importantly, suramin-mediated inhibition of RNA binding increased the expression of miR-1-3p, and enhanced sensitivity of cancer cells to Bcl-2 inhibitor navitoclax treatment. Taken together, we demonstrate as proof-of-concept a mechanism and an inhibitor compound for SND1 regulation of the survival of cancer cells through tumor suppressor miRNAs.",

keywords = "colon cancer, inhibitor screen, RNA binding protein, stress response",

author = "Saara Lehmusvaara and Teemu Haikarainen and Juha Saarikettu and Nieto, \{Guillermo Martinez\} and Olli Silvennoinen",

note = "Funding Information: Acknowledgments: The authors want to thank Antti Kurttila, Sanna Liukkonen and Merja Peltomaa for their valuable technical help. The authors also thank the Tampere facility of Virus Production for their service, and the DDCB core facility supported by the University of Helsinki and Biocenter Finland. The authors also thank Laura Turunen at the High Throughput Biomedicine Unit at the Institute for Molecular Medicine Finland (FIMM). Funding Information: Funding: This research was funded by the Academy of Finland (grant number 335437), Sigrid Jus{\'e}lius Foundation (grant number 1152), Finnish Cancer Foundation (grant number 4708167), Jane and Aatos Erkko Foundation, Tampere Tuberculosis Foundation (grant number 2019), and Pirkanmaa hospital district competitive research funding. Publisher Copyright: {\textcopyright} 2022 by the authors. Licensee MDPI, Basel, Switzerland.",

year = "2022",

month = jun,

doi = "10.3390/cancers14133100",

language = "English",

volume = "14",

journal = "Cancers",

issn = "2072-6694",

publisher = "Multidisciplinary Digital Publishing Institute (MDPI)",

number = "13",

}

TY - JOUR

T1 - Inhibition of RNA Binding in SND1 Increases the Levels of miR-1-3p and Sensitizes Cancer Cells to Navitoclax

AU - Lehmusvaara, Saara

AU - Haikarainen, Teemu

AU - Saarikettu, Juha

AU - Nieto, Guillermo Martinez

AU - Silvennoinen, Olli

N1 - Funding Information: Acknowledgments: The authors want to thank Antti Kurttila, Sanna Liukkonen and Merja Peltomaa for their valuable technical help. The authors also thank the Tampere facility of Virus Production for their service, and the DDCB core facility supported by the University of Helsinki and Biocenter Finland. The authors also thank Laura Turunen at the High Throughput Biomedicine Unit at the Institute for Molecular Medicine Finland (FIMM). Funding Information: Funding: This research was funded by the Academy of Finland (grant number 335437), Sigrid Jusélius Foundation (grant number 1152), Finnish Cancer Foundation (grant number 4708167), Jane and Aatos Erkko Foundation, Tampere Tuberculosis Foundation (grant number 2019), and Pirkanmaa hospital district competitive research funding. Publisher Copyright: © 2022 by the authors. Licensee MDPI, Basel, Switzerland.

PY - 2022/6

Y1 - 2022/6

N2 - SND1 is an RNA-binding protein overexpressed in large variety of cancers. SND1 has been proposed to enhance stress tolerance in cancer cells, but the molecular mechanisms are still poorly understood. We analyzed the expression of 372 miRNAs in the colon carcinoma cell line and show that SND1 silencing increases the expression levels of several tumor suppressor miRNAs. Furthermore, SND1 knockdown showed synergetic effects with cancer drugs through MEK-ERK and Bcl-2 family-related apoptotic pathways. To explore whether the SND1-mediated RNA bind-ing/degradation is responsible for the observed effect, we developed a screening assay to identify small molecules that inhibit the RNA-binding function of SND1. The screen identified P2X purinoreceptor antagonists as the most potent inhibitors. Validation confirmed that the best hit, suramin, inhibits the RNA binding ability of SND1. The binding characteristics and mode of suramin to SND1 were characterized biophysically and by molecular docking that identified positively charged binding cavities in Staphylococcus nuclease domains. Importantly, suramin-mediated inhibition of RNA binding increased the expression of miR-1-3p, and enhanced sensitivity of cancer cells to Bcl-2 inhibitor navitoclax treatment. Taken together, we demonstrate as proof-of-concept a mechanism and an inhibitor compound for SND1 regulation of the survival of cancer cells through tumor suppressor miRNAs.

AB - SND1 is an RNA-binding protein overexpressed in large variety of cancers. SND1 has been proposed to enhance stress tolerance in cancer cells, but the molecular mechanisms are still poorly understood. We analyzed the expression of 372 miRNAs in the colon carcinoma cell line and show that SND1 silencing increases the expression levels of several tumor suppressor miRNAs. Furthermore, SND1 knockdown showed synergetic effects with cancer drugs through MEK-ERK and Bcl-2 family-related apoptotic pathways. To explore whether the SND1-mediated RNA bind-ing/degradation is responsible for the observed effect, we developed a screening assay to identify small molecules that inhibit the RNA-binding function of SND1. The screen identified P2X purinoreceptor antagonists as the most potent inhibitors. Validation confirmed that the best hit, suramin, inhibits the RNA binding ability of SND1. The binding characteristics and mode of suramin to SND1 were characterized biophysically and by molecular docking that identified positively charged binding cavities in Staphylococcus nuclease domains. Importantly, suramin-mediated inhibition of RNA binding increased the expression of miR-1-3p, and enhanced sensitivity of cancer cells to Bcl-2 inhibitor navitoclax treatment. Taken together, we demonstrate as proof-of-concept a mechanism and an inhibitor compound for SND1 regulation of the survival of cancer cells through tumor suppressor miRNAs.

KW - colon cancer

KW - inhibitor screen

KW - RNA binding protein

KW - stress response

U2 - 10.3390/cancers14133100

DO - 10.3390/cancers14133100

M3 - Article

AN - SCOPUS:85132544819

SN - 2072-6694

VL - 14

JO - Cancers

JF - Cancers

IS - 13

M1 - 3100

ER -

Inhibition of RNA Binding in SND1 Increases the Levels of miR-1-3p and Sensitizes Cancer Cells to Navitoclax

Abstrakti

Rahoitus

YK:n kestävän kehityksen tavoitteet

Julkaisufoorumi-taso

!!ASJC Scopus subject areas

Pääsy asiakirjaan

Sormenjälki

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