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.
View: Text | PDF
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

×

Figure 4

αv Integrins mediate TGF-β1 release by GSCs and GSC-induced NK cell dysfunction.

Options: View larger image (or click on image) Download as PowerPoint
αv Integrins mediate TGF-β1 release by GSCs and GSC-induced NK cell dysf...
(A) Box-and-whisker plots showing total TGF-β (pg/mL) in the supernatant of NK cells and GSCs cultured either alone or together in the presence or absence of the αv integrin small molecule inhibitor cilengitide (10 μM) for 48 hours was determined by ELISA (n = 11). (B) Box-and-whisker plots showing MFI of p-Smad2/3 expression on HC-NK cells cultured either alone or with GSCs in the presence or absence of cilengitide (10 μM). (C) 51Cr release assay of K562 cell killing by NK cells cultured either alone or after coculture with GSCs for 48 hours in the presence or absence of cilengitide (10 μM) (n = 8). Red asterisks: specific lysis of K562 targets by NK cells that were cocultured with GSCs in the presence or absence cilengitide. (D and E) Representative zebra plots (D) and summary box-and-whisker plots (E) of CD107, IFN-γ, and TNF-α production by NK cells in response to K562 cells cultured either alone or after 48 hours of coculture with GSCs at a 1:1 ratio with or without cilengitide (n = 12). Inset numbers in panel D are the percentages of CD107a-, IFN-γ–, or TNF-α–positive NK cells within the indicated regions. (F) 51Cr release assay of K562 cell targets by NK cells cultured either alone or with WT GSCs or with CD51-KO GSCs for 48 hours at a 1:1 ratio (n = 3). Red asterisks: the specific lysis of K562 cell targets by NK cells after coculture with WT GSCs vs. CD51 KO. Statistical analysis by 2-way ANOVA with Bonferroni’s correction for multiple comparisons (A, B, E, and F) or 2-way ANOVA with Dunnett’s correction for multiple comparisons (C). *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001.