La Trobe

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Cancer stem cell marker DCLK1 reprograms small extracellular vesicles toward migratory phenotype in gastric cancer cells

Doublecortin-like kinase 1 (DCLK1) is a putative cancer stem cell marker, a promising diagnostic and prognostic maker for malignant tumors and a proposed driver gene for gastric cancer (GC). DCLK1 overexpression in a majority of solid cancers correlates with lymph node metastases, advanced disease and overall poor-prognosis. In cancer cells, DCLK1 expression has been shown to promote epithelial-to-mesenchymal transition (EMT), driving disruption of cell-cell adhesion, cell migration and invasion. Here, we report that DCLK1 influences small extracellular vesicle (sEV/exosome) biogenesis in a kinase-dependent manner. sEVs isolated from DCLK1 overexpressing human GC cell line MKN1 (MKN1OE-sEVs), promote the migration of parental (non-transfected) MKN1 cells (MKN1PAR). Quantitative proteome analysis of MKN1OE-sEVs revealed enrichment in migratory and adhesion regulators (STRAP, CORO1B, BCAM, COL3A, CCN1) in comparison to MKN1PAR-sEVs. Moreover, using DCLK1-IN-1, a specific small molecule inhibitor of DCLK1, we reversed the increase in sEV size and concentration in contrast to other EV subtypes, as well as kinase-dependent cargo selection of proteins involved in EV biogenesis (KTN1, CHMP1A, MYO1G) and migration and adhesion processes (STRAP, CCN1). Our findings highlight a specific role of DCLK1-kinase dependent cargo selection for sEVs and shed new light on its role as a regulator of signaling in gastric tumorigenesis.

Funding

HelenAmelia Hains Fellowship; DF/HCCGI SPOREDevelopmental Research Project Award, Grant/Award Number: P50CA127003; HaleCenter forPancreaticResearch; La Trobe University, Grant/Award Numbers: RFAUnderstandingDisease grant, postgraduate research fellowship LTUPRS; Dana-Farber/Harvard CancerCenter, Grant/Award Number: DF/HCC GI SPORE/P50CA127003; National Health and Medical Research Council (NHMRC), Grant/Award Numbers: Investigator Grant/1183814, ProgramGrant/1092788, Project Grant/1139489, Senior Research Fellowship/1079257; StateGovernment ofVictoria, Grant/Award Number: Operational Infrastructure Support Program

History

Publication Date

01/07/2021

Journal

Proteomics

Volume

21

Issue

13-14

Article Number

2000098

Pagination

16p.

Publisher

Wiley

ISSN

1615-9853

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