Intercellular transfer of oncogenic KRAS via tunneling nanotubes introduces intracellular mutational heterogeneity in colon cancer cells

Snider Desir, Phillip Wong, Thomas Turbyville, De Chen, Mihir Shetty, Christopher Clark, Edward Zhai, Yevgeniy Romin, Katia Manova-Todorova, Tim Starr, Dwight V. Nissley, Clifford J Steer, Subree Subramanian, Emil Lou

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Mutated forms of the RAS oncogene drive 30% of all cancers, but they cannot be targeted therapeutically using currently available drugs. The molecular and cellular mechanisms that create a heterogenous tumor environment harboring both mutant and wild-type RAS have not been elucidated. In this study, we examined horizontal transfer of mutant KRAS (Kirsten Rat Sarcoma Virus) between colorectal cancer (CRC) cells via a direct form of cell-to-cell communication called tunneling nanotubes (TNTs). TNT formation was significantly higher in CRC cell lines expressing mutant KRAS than CRC cell lines expressing wild-type RAS; this effect was most pronounced in metastatic CRC cell lines with both mutant KRAS and deficiency in mismatch repair proteins. Using inverted and confocal fluorescence time-lapse and fluorescence recovery after photobleaching (FRAP)-based microscopy, we observed GFP-tagged mutant KRASG12D protein trafficking between CRC cells through TNTs within a span of seconds to several minutes. Notably, acquisition of mutant KRAS increased Extracellular Signal-regulated Kinase (ERK) phosphorylation and upregulated tunneling nanotube formation in recipient wildtype CRC cells. In conclusion, these findings suggest that intercellular horizontal transfer of RAS can occur by TNTs. We propose that intercellular transfer of mutant RAS can potentially induce intratumoral heterogeneity and result in a more invasive phenotype in recipient cells.

Original languageEnglish (US)
Article number892
JournalCancers
Volume11
Issue number7
DOIs
StatePublished - Jul 1 2019

Fingerprint

Kirsten murine sarcoma virus
Nanotubes
Oncogenic Viruses
Parvovirus
Colonic Neoplasms
Colorectal Neoplasms
Cell Line
Fluorescence Recovery After Photobleaching
Extracellular Signal-Regulated MAP Kinases
Protein Transport
Mutant Proteins
Oncogenes
Cell Communication
Microscopy
Neoplasms
Fluorescence
Phosphorylation
Phenotype

Keywords

  • Cellular reprogramming
  • Colon cancer
  • Colorectal cancer
  • Confocal microscopy
  • Intercellular communication
  • Intercellular transfer
  • KRAS
  • Oncogene
  • Tunneling nanotubes

PubMed: MeSH publication types

  • Journal Article

Cite this

Intercellular transfer of oncogenic KRAS via tunneling nanotubes introduces intracellular mutational heterogeneity in colon cancer cells. / Desir, Snider; Wong, Phillip; Turbyville, Thomas; Chen, De; Shetty, Mihir; Clark, Christopher; Zhai, Edward; Romin, Yevgeniy; Manova-Todorova, Katia; Starr, Tim; Nissley, Dwight V.; Steer, Clifford J; Subramanian, Subree; Lou, Emil.

In: Cancers, Vol. 11, No. 7, 892, 01.07.2019.

Research output: Contribution to journalArticle

Desir, Snider ; Wong, Phillip ; Turbyville, Thomas ; Chen, De ; Shetty, Mihir ; Clark, Christopher ; Zhai, Edward ; Romin, Yevgeniy ; Manova-Todorova, Katia ; Starr, Tim ; Nissley, Dwight V. ; Steer, Clifford J ; Subramanian, Subree ; Lou, Emil. / Intercellular transfer of oncogenic KRAS via tunneling nanotubes introduces intracellular mutational heterogeneity in colon cancer cells. In: Cancers. 2019 ; Vol. 11, No. 7.
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AU - Clark, Christopher

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AB - Mutated forms of the RAS oncogene drive 30% of all cancers, but they cannot be targeted therapeutically using currently available drugs. The molecular and cellular mechanisms that create a heterogenous tumor environment harboring both mutant and wild-type RAS have not been elucidated. In this study, we examined horizontal transfer of mutant KRAS (Kirsten Rat Sarcoma Virus) between colorectal cancer (CRC) cells via a direct form of cell-to-cell communication called tunneling nanotubes (TNTs). TNT formation was significantly higher in CRC cell lines expressing mutant KRAS than CRC cell lines expressing wild-type RAS; this effect was most pronounced in metastatic CRC cell lines with both mutant KRAS and deficiency in mismatch repair proteins. Using inverted and confocal fluorescence time-lapse and fluorescence recovery after photobleaching (FRAP)-based microscopy, we observed GFP-tagged mutant KRASG12D protein trafficking between CRC cells through TNTs within a span of seconds to several minutes. Notably, acquisition of mutant KRAS increased Extracellular Signal-regulated Kinase (ERK) phosphorylation and upregulated tunneling nanotube formation in recipient wildtype CRC cells. In conclusion, these findings suggest that intercellular horizontal transfer of RAS can occur by TNTs. We propose that intercellular transfer of mutant RAS can potentially induce intratumoral heterogeneity and result in a more invasive phenotype in recipient cells.

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