Dependency of human and murine LKB1-inactivated lung cancer on aberrant CRTC-CREB activation

  1. Xin Zhou
  2. Jennifer W Li
  3. Zirong Chen
  4. Wei Ni
  5. Xuehui Li
  6. Rongqiang Yang
  7. Huangxuan Shen
  8. Jian Liu
  9. Franco J DeMayo
  10. Jianrong Lu
  11. Frederic J Kaye
  12. Lizi Wu  Is a corresponding author
  1. University of Florida, United States
  2. Sun Yat-sen University, China
  3. Zhejiang University, China
  4. NIH, United States
  5. University of Florida College of Medicine, United States

Abstract

Lung cancer with loss-of-function of the LKB1 tumor suppressor is a common aggressive subgroup with no effective therapies. LKB1-deficiency induces constitutive activation of cAMP/CREB-mediated transcription by a family of three CREB-regulated transcription coactivators (CRTC1-3). However, the significance and mechanism of CRTC activation in promoting the aggressive phenotype of LKB1-null cancer remain poorly characterized. Here we observed overlapping CRTC expression patterns and mild growth phenotypes of individual CRTC-knockouts in lung cancer, suggesting functional redundancy of CRTC1-3. We consequently designed a dominant-negative mutant (dnCRTC) to block all three CRTCs to bind and co-activate CREB. Expression of dnCRTC efficiently inhibited the aberrantly activated cAMP/CREB-mediated oncogenic transcriptional program induced by LKB1-deficiency, and specifically blocked the growth of human and murine LKB1-inactivated lung cancer. Collectively, this study provides direct proof for an essential role of the CRTC-CREB activation in promoting the malignant phenotypes of LKB1-null lung cancer and proposes the CRTC-CREB interaction interface as a novel therapeutic target.

Data availability

The transcriptomic data were deposited in the NCBI GEO database GSE157722.All data generated or analyzed for this study are included in the manuscript.

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Xin Zhou

    Molecular Genetics & Microbiology, University of Florida, Gainesville, United States
    Competing interests
    The authors declare that no competing interests exist.
  2. Jennifer W Li

    Department of Medicine, University of Florida, Gainesville, United States
    Competing interests
    The authors declare that no competing interests exist.
  3. Zirong Chen

    Molecular Genetics & Microbiology, University of Florida, Gainesville, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Wei Ni

    Molecular Genetics & Microbiology, University of Florida, Gainesville, United States
    Competing interests
    The authors declare that no competing interests exist.
  5. Xuehui Li

    Molecular Genetics & Microbiology, University of Florida, Gainesville, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Rongqiang Yang

    Molecular Genetics & Microbiology, University of Florida, Gainesville, United States
    Competing interests
    The authors declare that no competing interests exist.
  7. Huangxuan Shen

    Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China
    Competing interests
    The authors declare that no competing interests exist.
  8. Jian Liu

    ZJU-UoE Institute, Zhejiang University, Haining, China
    Competing interests
    The authors declare that no competing interests exist.
  9. Franco J DeMayo

    NIH, Research Triangle Park, United States
    Competing interests
    The authors declare that no competing interests exist.
  10. Jianrong Lu

    Biochemistry and Molecular Biology, University of Florida College of Medicine, Gainesville, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4969-6040
  11. Frederic J Kaye

    Medicine, University of Florida, Gainesville, United States
    Competing interests
    The authors declare that no competing interests exist.
  12. Lizi Wu

    Molecular Genetics & Microbiology, University of Florida, Gainesville, United States
    For correspondence
    lzwu@ufl.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0076-2617

Funding

National Cancer Institute (R01CA234351)

  • Lizi Wu

National Institute of Dental and Craniofacial Research (R01DE023641)

  • Lizi Wu

UF Health Cancer Center

  • Lizi Wu

National Institute of Environmental Health Sciences (Z1AES103311-01)

  • Franco J DeMayo

University of Florida Gatorade Trust

  • Frederic J Kaye

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: Animal studies were performed following a protocol approved by the IACUC (Institutional Animal Care & Use Committee) of the University of Florida (201810386). All animals were housed, cared for, and used in an animal care facility at the University of Florida that is fully accredited by the Association for the Assessment and Accreditation of Laboratory Animal Care International (AAALAC) program in compliance with the Guide for the Care and Use of Laboratory Animals, the Animal Welfare Act and other applicable state and local regulations.

Reviewing Editor

  1. Erica A Golemis, Fox Chase Cancer Center, United States

Publication history

  1. Received: December 29, 2020
  2. Accepted: June 17, 2021
  3. Accepted Manuscript published: June 18, 2021 (version 1)
  4. Version of Record published: June 28, 2021 (version 2)
  5. Version of Record updated: July 2, 2021 (version 3)

Copyright

This is an open-access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.

Metrics

  • 1,402
    Page views
  • 200
    Downloads
  • 2
    Citations

Article citation count generated by polling the highest count across the following sources: PubMed Central, Crossref, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Xin Zhou
  2. Jennifer W Li
  3. Zirong Chen
  4. Wei Ni
  5. Xuehui Li
  6. Rongqiang Yang
  7. Huangxuan Shen
  8. Jian Liu
  9. Franco J DeMayo
  10. Jianrong Lu
  11. Frederic J Kaye
  12. Lizi Wu
(2021)
Dependency of human and murine LKB1-inactivated lung cancer on aberrant CRTC-CREB activation
eLife 10:e66095.
https://doi.org/10.7554/eLife.66095

Further reading

    1. Cancer Biology
    2. Cell Biology
    Johnny M Tkach et al.
    Research Article

    Centrosomes act as the main microtubule organizing center (MTOC) in metazoans. Centrosome number is tightly regulated by limiting centriole duplication to a single round per cell cycle. This control is achieved by multiple mechanisms, including the regulation of the protein kinase PLK4, the most upstream facilitator of centriole duplication. Altered centrosome numbers in mouse and human cells cause p53-dependent growth arrest through poorly defined mechanisms. Recent work has shown that the E3 ligase TRIM37 is required for cell cycle arrest in acentrosomal cells. To gain additional insights into this process, we undertook a series of genome-wide CRISPR/Cas9 screens to identify factors important for growth arrest triggered by treatment with centrinone B, a selective PLK4 inhibitor. We found that TRIM37 is a key mediator of growth arrest after partial or full PLK4 inhibition. Interestingly, PLK4 cellular mobility decreased in a dose-dependent manner after centrinone B treatment. In contrast to recent work, we found that growth arrest after PLK4 inhibition correlated better with PLK4 activity than with mitotic length or centrosome number. These data provide insights into the global response to changes in centrosome number and PLK4 activity and extend the role for TRIM37 in regulating the abundance, localization, and function of centrosome proteins.

    1. Cancer Biology
    2. Cell Biology
    Haoran Zhu et al.
    Research Article

    Hyperactivation of oncogenic pathways downstream of RAS and PI3K/AKT in normal cells induces a senescence-like phenotype that acts as a tumor-suppressive mechanism that must be overcome during transformation. We previously demonstrated that AKT-induced senescence (AIS) is associated with profound transcriptional and metabolic changes. Here, we demonstrate that human fibroblasts undergoing AIS display upregulated cystathionine-β-synthase (CBS) expression and enhanced uptake of exogenous cysteine, which lead to increased hydrogen sulfide (H2S) and glutathione (GSH) production, consequently protecting senescent cells from oxidative stress-induced cell death. CBS depletion allows AIS cells to escape senescence and re-enter the cell cycle, indicating the importance of CBS activity in maintaining AIS. Mechanistically, we show this restoration of proliferation is mediated through suppressing mitochondrial respiration and reactive oxygen species (ROS) production by reducing mitochondrial localized CBS while retaining antioxidant capacity of transsulfuration pathway. These findings implicate a potential tumor-suppressive role for CBS in cells with aberrant PI3K/AKT pathway activation. Consistent with this concept, in human gastric cancer cells with activated PI3K/AKT signaling, we demonstrate that CBS expression is suppressed due to promoter hypermethylation. CBS loss cooperates with activated PI3K/AKT signaling in promoting anchorage-independent growth of gastric epithelial cells, while CBS restoration suppresses the growth of gastric tumors in vivo. Taken together, we find that CBS is a novel regulator of AIS and a potential tumor suppressor in PI3K/AKT-driven gastric cancers, providing a new exploitable metabolic vulnerability in these cancers.