Targeting the αv integrin/TGF-β axis improves natural killer cell function against glioblastoma stem cells
Hila Shaim, … , Amy B. Heimberger, Katayoun Rezvani
Hila Shaim, … , Amy B. Heimberger, Katayoun Rezvani
Published June 17, 2021
Citation Information: J Clin Invest. 2021;131(14):e142116. https://doi.org/10.1172/JCI142116.
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Research Article Immunology

Targeting the αv integrin/TGF-β axis improves natural killer cell function against glioblastoma stem cells

Abstract

Glioblastoma multiforme (GBM), the most aggressive brain cancer, recurs because glioblastoma stem cells (GSCs) are resistant to all standard therapies. We showed that GSCs, but not normal astrocytes, are sensitive to lysis by healthy allogeneic natural killer (NK) cells in vitro. Mass cytometry and single-cell RNA sequencing of primary tumor samples revealed that GBM tumor–infiltrating NK cells acquired an altered phenotype associated with impaired lytic function relative to matched peripheral blood NK cells from patients with GBM or healthy donors. We attributed this immune evasion tactic to direct cell-to-cell contact between GSCs and NK cells via αv integrin–mediated TGF-β activation. Treatment of GSC-engrafted mice with allogeneic NK cells in combination with inhibitors of integrin or TGF-β signaling or with TGFBR2 gene–edited allogeneic NK cells prevented GSC-induced NK cell dysfunction and tumor growth. These findings reveal an important mechanism of NK cell immune evasion by GSCs and suggest the αv integrin/TGF-β axis as a potentially useful therapeutic target in GBM.

Authors

Hila Shaim, Mayra Shanley, Rafet Basar, May Daher, Joy Gumin, Daniel B. Zamler, Nadima Uprety, Fang Wang, Yuefan Huang, Konrad Gabrusiewicz, Qi Miao, Jinzhuang Dou, Abdullah Alsuliman, Lucila N. Kerbauy, Sunil Acharya, Vakul Mohanty, Mayela Mendt, Sufang Li, JunJun Lu, Jun Wei, Natalie W. Fowlkes, Elif Gokdemir, Emily L. Ensley, Mecit Kaplan, Cynthia Kassab, Li Li, Gonca Ozcan, Pinaki P. Banerjee, Yifei Shen, April L. Gilbert, Corry M. Jones, Mustafa Bdiwi, Ana K. Nunez-Cortes, Enli Liu, Jun Yu, Nobuhiko Imahashi, Luis Muniz-Feliciano, Ye Li, Jian Hu, Giulio Draetta, David Marin, Dihua Yu, Stephan Mielke, Matthias Eyrich, Richard E. Champlin, Ken Chen, Frederick F. Lang, Elizabeth J. Shpall, Amy B. Heimberger, Katayoun Rezvani

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Figure 3

GSC-induced NK cell dysfunction requires cell-cell contact.

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GSC-induced NK cell dysfunction requires cell-cell contact. (A) p-Smad2/...
(A) p-Smad2/3 (MFI) expression in NK cells cultured alone or with GSCs in the presence or absence of LY2109761 or galunisertib (n = 4). (B) HC-NK cells were cultured with or without GSCs for 48 hours in the presence or absence of LY2109761 or galunisertib. A 4-hour 51Cr release assay tested their cytotoxicity against K562 cell (left) or GSC (right) targets. Asterisks represent the statistical difference in NK cell cytotoxicity in the presence or absence of galunisertib (gray) or LY2109761 (black) (n = 3). (C) TI-NK cells were cultured overnight with or without galunisertib and their cytotoxicity tested against K562 cell targets in a 4-hour 51Cr release assay. Black asterisks: TI-NK + galunisertib vs. GP-NK (n = 3). (D and E) Total TGF-β1 (pg/mL; ELISA) levels in supernatants from NK cells and GSCs cultured alone or together for 48 hours in direct contact or separated with a Transwell membrane (D; n = 13) or NK cells and astrocytes cultured alone or together for 48 hours (E; n = 3). (F) NK cells cocultured with GSCs for 48 hours in direct contact or separated with a Transwell and their cytotoxicity tested against K562 in a 4-hour 51Cr release assay (n = 7). (G) p-Smad2/3 (MFI) expression in HC-NK cells cultured overnight with or without GSCs in the presence or absence of TGF-β–blocking antibodies, or separated with a Transwell membrane (n = 5). (H) Total TGF-β1 (ELISA) in the supernatant of NK cells and GSCs cultured alone (NK: blue; GSC: black) or together (red) (n = 4). Blue asterisks: GSCs vs. NK:GSCs (E:T). (I) Fold-change in TGFB1 mRNA levels in NK cells and GSCs cultured for 48 hours either alone, or together in direct contact or separated with a Transwell membrane (n = 7). Statistical analysis by 2-way ANOVA with Dunnett’s (A–C and E–H), Tukey’s (D), or Bonferroni’s (I) correction for multiple comparisons. *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001.