Close

An NKX2-1/ERK/WNT feedback loop modulates gastric identity and response to targeted therapy in lung adenocarcinoma

Rediet Zewdu, Elnaz Mirzaei Mehrabad, Kelley Ingram, Alex Jones, Soledad A. Camolotto, Michelle C. Mendoza, Benjamin Spike, Eric L. Snyder

Preprint posted on 10 May 2020 https://www.biorxiv.org/content/10.1101/2020.02.25.965004v1

Article now published in eLife at http://dx.doi.org/10.7554/eLife.66788

Identifying gastric-like cells in primary lung cancer: Zewdu et al. observe a shift in gastric-like cell signatures after targeted therapy of invasive mucinous adenocarcinoma.

Selected by Catherine Dabrowska

Background

Lung adenocarcinoma (LuAD) is the most common cause of cancer deaths worldwide (Herbst et al., 2018). LuAD itself accounts for multiple subtypes, including invasive mucinous adenocarcinoma (IMA) (Cha & Shim, 2017). Fascinatingly, some primary IMAs of the lung have been observed to contain cells which no longer express lung fate lineage markers. How is possible that some cells of a primary lung tumour can lose expression of lung markers? The answer can be found by examining which molecular pathways are critical for specifying the lung tissue during development. In the embryo, the lung is portioned off from the foregut endoderm by expression of Nkx2.1, a master transcription factor determining the lung lineage (Faure & De Santa Barbara, 2011). The foregut endoderm also gives rise to organs of the gastrointestinal tract, including the stomach (Faure & De Santa Barbara, 2011). The corpus epithelium of the adult stomach is invaginated into glands responsible for producing stomach acid and digestive enzymes. The cells constituting these glands are renewed by the function of resident stem and progenitor cells respectively found at the base and isthmus of the glands (Han et al., 2019). Multiple research groups have now shown that loss of Nkx2.1 in mouse primary lung tumours driven by Kras mutation can result in some cells acquiring gastric fate lineage markers (Snyder et al., 2013; Tata et al., 2018). BRaf is also found to be mutated in LuAD IMA subtypes (Cha & Shim, 2017). Here, Zewdu et al. go a step further and begin to consider the implications of targeted therapy on lineage switching within gastric subtypes.

Key findings

NKX2.1 levels determine tumour morphology and cellular subtypes
The authors utilised two murine models (1) BRafV600E/+ ;Trp53f/f; Nkx2-1f/+; Rosa26LSL-tdTomato (BP) mice and (2) BRafV600E/+; Trp53f/f; Nkx2-1f/f; Rosa26LSL-tdTomato (BPN) mice. The tumours formed in the BPN model morphologically resembled human IMAs and also were found to have increase in gastric markers and loss of surfactant protein C, a key alveolar marker. This confirmed previous findings in published literature that loss of Nkx2.1 can convert cells in LuADs from pulmonary fate to gastric fate.

 

NKX2-1 regulates MAPK pathway in BRAFV600E-driven LUAD
In order to drive tumours in mice which were physiologically comparable to how tumours develop in humans, the authors established mouse lines which first developed LuAD due to combined Trp53 homozygous knockout and BRafV600E/+ mutation. Post tumour growth, the authors then removed Nkx2.1 expression by utilising floxed alleles under the control of Rosa26-CreERT2. Compared to controls where Nkx2.1 expression remained at wild-type levels, Nkx2.1 deletion in mice induced tumour transition to the IMA subtype, as expected. Survival between the two experimental groups was not significantly different. The authors detected raised phosphorylated ERK signaling in the tumours lacking Nkx2.1 expression and also identified strong pERK expression in human IMA.

 

Response of NKX2-1-positive and NKX2-1-negative tumors to RAF/MEK inhibition
Following from their identification of increased pERK expression in the step-wise models, the authors sought to chemically target the RAF/MEK pathway. The combined administration of PLX4720 (BRAFV600E inhibitor) and PD0325901 (MEK inhibitor) resulted in suppression of ERK phosphorylation in both BP and BPN mice, with a corresponding greater decrease of tumour burden in the BPN model, as expected. This indicated greater sensitivity to MEK inhibition in the Nkx2.1 negative mice. However, upon cessation of inhibitor treatment residual cells rapidly reformed tumours. The authors further sought to understand these differences by utilising single cell RNA sequencing to compare treated and untreated BP and BPN mice. This experiment suggested that residual cells within treated BPN tumours retained their ability to remain in the cell cycle and so the authors caution of the necessity for consideration of distinct therapeutic strategies to target these tumours.

 

MAPK pathway regulates identity of NKX2-1-negative mucinous adenocarcinoma within the gastric lineage
Critically, the authors delve further into their RNA sequencing datasets and observe that there is a shift in gastric marker expression when BPN tumours are subjected to MAPK inhibition compared to no treatment control. BPN tumours with no treatment highly expressed transcripts belonging to mucous cells, e.g. Muc5ac. After treatment however, these transcripts become downregulated and instead pepsinogen C and chymosin, markers of the chief cells found at the base of the gastric glands, become upregulated (see bulk RNA sequencing figure A, taken from Figure 6A of the preprint). This was further supported by the enrichment of the Lgr5-expressing chief cell-like signature. In addition, Tff2 was found to be upregulated, a marker of the isthmus progenitor cells of the gastric glandular epithelium. The authors confirmed the pepsinogen C finding with immunohistochemistry of untreated and treated BP and BPN tumours. As Wnt signalling is known to be enriched at the base of the stomach glands, the authors searched their dataset and identified that after treatment of BPN tumours, Wnt targets were upregulated. By supplying Wnt pathway inhibitors to established organoids derived from BPN tumours, they were able to show the cultures could support tuft cell identity but could not support chief cells.

 

A) MAPK inhibition of BPN tumours (BPN Tx) causes upregulation of a Chief cell-like profile. (Taken from Figure 6A of the preprint).

Why I chose this preprint

A recent trend in scientific thought has been to consider how the molecular pathways critical for organ specification in development can be perturbed in tumourigenesis, tumour progression and therapeutic evasion. This paper went beyond simply observing a switch to a gastric phenotype and instead identified that after MAPK inhibition treatment in tumours negative for Nkx2.1, there was a shift in the gastric cell expression profile itself. Critically, the authors observed and detailed how MAPK inhibition could drive cells in IMA tumours in the lung from having a gastric mucous expression profile to a gastric stem and progenitor expression profile.

Questions for the authors

The authors detailed some interesting observations. These could be improved upon with more in-depth analysis of the single cell sequencing dataset, in order to provide more evidence of the claimed fate change, and subsequent focused in vitro studies. Organoid models are very useful tools for testing various hypotheses, which the authors did utilise in part of their study.

  1. The authors show, using organoid culture, the reliance of the IMA ‘chief cells’ on Wnt signaling, as in native, true stomach chief cells. Can the authors demonstrate the cellular source of the Wnt in the IMA?
  2. Do the authors see a difference in the stromal and immune cell profile between untreated and treated BP and BPN tumours? Are particular cells excluded or included during the switch to gastric fate, and what is the implication of this on the surrounding lung tissue? These questions can be examined utilising organoid co-cultures.

References

Cha, Y. J., & Shim, H. S. (2017, October 1). Biology of invasive mucinous adenocarcinoma of the lung. Translational Lung Cancer Research. AME Publishing Company. https://doi.org/10.21037/tlcr.2017.06.10

Faure, S., & De Santa Barbara, P. (2011). Molecular embryology of the foregut. Journal of Pediatric Gastroenterology and Nutrition, 52(SUPPL. 1), S2. https://doi.org/10.1097/MPG.0b013e3182105a1a

Han, S., Fink, J., Jörg, D. J., Lee, E., Yum, M. K., Chatzeli, L., … Koo, B.-K. (2019). Defining the Identity and Dynamics of Adult Gastric Isthmus Stem Cells. Cell Stem Cell, 25(3), 342-356.e7. https://doi.org/10.1016/j.stem.2019.07.008

Herbst, R. S., Morgensztern, D., & Boshoff, C. (2018, January 24). The biology and management of non-small cell lung cancer. Nature. Nature Publishing Group. https://doi.org/10.1038/nature25183

Snyder, E. L., Watanabe, H., Magendantz, M., Hoersch, S., Chen, T. A., Wang, D. G., … Jacks, T. (2013). Nkx2-1 Represses a Latent Gastric Differentiation Program in Lung Adenocarcinoma. Molecular Cell, 50(2), 185–199. https://doi.org/10.1016/j.molcel.2013.02.018

Tata, P. R., Chow, R. D., Saladi, S. V., Kimura, S., & Ellisen, L. W. (2018). Developmental History Provides a Roadmap for the Emergence of Tumor Plasticity. Developmental Cell, 44, 679-693.e5. https://doi.org/10.1016/j.devcel.2018.02.024

 

Tags: adenocarcinoma, cancer, chief cell, isthmus cell, lung, mouse

Posted on: 10 May 2020 , updated on: 14 May 2020

doi: https://doi.org/10.1242/prelights.20357

Read preprint (No Ratings Yet)

Have your say

Your email address will not be published. Required fields are marked *

This site uses Akismet to reduce spam. Learn how your comment data is processed.

Sign up to customise the site to your preferences and to receive alerts

Register here

preLists in the cancer biology category:

CSHL 87th Symposium: Stem Cells

Preprints mentioned by speakers at the #CSHLsymp23

 



List by Alex Eve

Journal of Cell Science meeting ‘Imaging Cell Dynamics’

This preList highlights the preprints discussed at the JCS meeting 'Imaging Cell Dynamics'. The meeting was held from 14 - 17 May 2023 in Lisbon, Portugal and was organised by Erika Holzbaur, Jennifer Lippincott-Schwartz, Rob Parton and Michael Way.

 



List by Helen Zenner

CellBio 2022 – An ASCB/EMBO Meeting

This preLists features preprints that were discussed and presented during the CellBio 2022 meeting in Washington, DC in December 2022.

 



List by Nadja Hümpfer et al.

Fibroblasts

The advances in fibroblast biology preList explores the recent discoveries and preprints of the fibroblast world. Get ready to immerse yourself with this list created for fibroblasts aficionados and lovers, and beyond. Here, my goal is to include preprints of fibroblast biology, heterogeneity, fate, extracellular matrix, behavior, topography, single-cell atlases, spatial transcriptomics, and their matrix!

 



List by Osvaldo Contreras

Single Cell Biology 2020

A list of preprints mentioned at the Wellcome Genome Campus Single Cell Biology 2020 meeting.

 



List by Alex Eve

ASCB EMBO Annual Meeting 2019

A collection of preprints presented at the 2019 ASCB EMBO Meeting in Washington, DC (December 7-11)

 



List by Madhuja Samaddar et al.

Lung Disease and Regeneration

This preprint list compiles highlights from the field of lung biology.

 



List by Rob Hynds

Anticancer agents: Discovery and clinical use

Preprints that describe the discovery of anticancer agents and their clinical use. Includes both small molecules and macromolecules like biologics.

 



List by Zhang-He Goh

Biophysical Society Annual Meeting 2019

Few of the preprints that were discussed in the recent BPS annual meeting at Baltimore, USA

 



List by Joseph Jose Thottacherry

Also in the cell biology category:

‘In preprints’ from Development 2022-2023

A list of the preprints featured in Development's 'In preprints' articles between 2022-2023

 



List by Alex Eve, Katherine Brown

preLights peer support – preprints of interest

This is a preprint repository to organise the preprints and preLights covered through the 'preLights peer support' initiative.

 



List by preLights peer support

The Society for Developmental Biology 82nd Annual Meeting

This preList is made up of the preprints discussed during the Society for Developmental Biology 82nd Annual Meeting that took place in Chicago in July 2023.

 



List by Joyce Yu, Katherine Brown

CSHL 87th Symposium: Stem Cells

Preprints mentioned by speakers at the #CSHLsymp23

 



List by Alex Eve

Journal of Cell Science meeting ‘Imaging Cell Dynamics’

This preList highlights the preprints discussed at the JCS meeting 'Imaging Cell Dynamics'. The meeting was held from 14 - 17 May 2023 in Lisbon, Portugal and was organised by Erika Holzbaur, Jennifer Lippincott-Schwartz, Rob Parton and Michael Way.

 



List by Helen Zenner

9th International Symposium on the Biology of Vertebrate Sex Determination

This preList contains preprints discussed during the 9th International Symposium on the Biology of Vertebrate Sex Determination. This conference was held in Kona, Hawaii from April 17th to 21st 2023.

 



List by Martin Estermann

Alumni picks – preLights 5th Birthday

This preList contains preprints that were picked and highlighted by preLights Alumni - an initiative that was set up to mark preLights 5th birthday. More entries will follow throughout February and March 2023.

 



List by Sergio Menchero et al.

CellBio 2022 – An ASCB/EMBO Meeting

This preLists features preprints that were discussed and presented during the CellBio 2022 meeting in Washington, DC in December 2022.

 



List by Nadja Hümpfer et al.

Fibroblasts

The advances in fibroblast biology preList explores the recent discoveries and preprints of the fibroblast world. Get ready to immerse yourself with this list created for fibroblasts aficionados and lovers, and beyond. Here, my goal is to include preprints of fibroblast biology, heterogeneity, fate, extracellular matrix, behavior, topography, single-cell atlases, spatial transcriptomics, and their matrix!

 



List by Osvaldo Contreras

EMBL Synthetic Morphogenesis: From Gene Circuits to Tissue Architecture (2021)

A list of preprints mentioned at the #EESmorphoG virtual meeting in 2021.

 



List by Alex Eve

FENS 2020

A collection of preprints presented during the virtual meeting of the Federation of European Neuroscience Societies (FENS) in 2020

 



List by Ana Dorrego-Rivas

Planar Cell Polarity – PCP

This preList contains preprints about the latest findings on Planar Cell Polarity (PCP) in various model organisms at the molecular, cellular and tissue levels.

 



List by Ana Dorrego-Rivas

BioMalPar XVI: Biology and Pathology of the Malaria Parasite

[under construction] Preprints presented at the (fully virtual) EMBL BioMalPar XVI, 17-18 May 2020 #emblmalaria

 



List by Dey Lab, Samantha Seah

1

Cell Polarity

Recent research from the field of cell polarity is summarized in this list of preprints. It comprises of studies focusing on various forms of cell polarity ranging from epithelial polarity, planar cell polarity to front-to-rear polarity.

 



List by Yamini Ravichandran

TAGC 2020

Preprints recently presented at the virtual Allied Genetics Conference, April 22-26, 2020. #TAGC20

 



List by Maiko Kitaoka et al.

3D Gastruloids

A curated list of preprints related to Gastruloids (in vitro models of early development obtained by 3D aggregation of embryonic cells). Updated until July 2021.

 



List by Paul Gerald L. Sanchez and Stefano Vianello

ECFG15 – Fungal biology

Preprints presented at 15th European Conference on Fungal Genetics 17-20 February 2020 Rome

 



List by Hiral Shah

ASCB EMBO Annual Meeting 2019

A collection of preprints presented at the 2019 ASCB EMBO Meeting in Washington, DC (December 7-11)

 



List by Madhuja Samaddar et al.

EMBL Seeing is Believing – Imaging the Molecular Processes of Life

Preprints discussed at the 2019 edition of Seeing is Believing, at EMBL Heidelberg from the 9th-12th October 2019

 



List by Dey Lab

Autophagy

Preprints on autophagy and lysosomal degradation and its role in neurodegeneration and disease. Includes molecular mechanisms, upstream signalling and regulation as well as studies on pharmaceutical interventions to upregulate the process.

 



List by Sandra Malmgren Hill

Lung Disease and Regeneration

This preprint list compiles highlights from the field of lung biology.

 



List by Rob Hynds

Cellular metabolism

A curated list of preprints related to cellular metabolism at Biorxiv by Pablo Ranea Robles from the Prelights community. Special interest on lipid metabolism, peroxisomes and mitochondria.

 



List by Pablo Ranea Robles

BSCB/BSDB Annual Meeting 2019

Preprints presented at the BSCB/BSDB Annual Meeting 2019

 



List by Dey Lab

MitoList

This list of preprints is focused on work expanding our knowledge on mitochondria in any organism, tissue or cell type, from the normal biology to the pathology.

 



List by Sandra Franco Iborra

ASCB/EMBO Annual Meeting 2018

This list relates to preprints that were discussed at the recent ASCB conference.

 



List by Dey Lab, Amanda Haage
Close