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[Killing effect of Robo1 targeted Chimeric Antigen Receptor modified NK92 cells against glioma and neuroblastoma

Y Qu1, J Z Bi

  • 1Department of Neural Medicine, Second Hospital of Shandong University, Jinan 250033, China.

Zhonghua Yi Xue Za Zhi
|April 3, 2018
PubMed

Insights

Robo1-CAR-NK92 cells demonstrate enhanced targeted killing of glioma and neuroblastoma cells. A combination of IL-15 and dexamethasone optimizes Robo1 CAR NK cell culture for improved proliferation and survival.

Area of Science:

  • Immunology
  • Cell Biology
  • Cancer Therapy

Background:

  • Natural Killer (NK) cells are crucial for innate immunity and cancer surveillance.
  • CAR-NK92 cells engineered with chimeric antigen receptors (CARs) offer targeted cancer immunotherapy.
  • Robo1 is a cell surface receptor implicated in various cellular processes.

Purpose of the Study:

  • To evaluate the cytotoxicity of Robo1-CAR-NK92 cells against U87-MG (glioblastoma) and SH-SY5Y (neuroblastoma) cells.
  • To investigate the impact of IL-15, IL-21, and dexamethasone on Robo1-CAR-NK92 cell functions.
  • To optimize the culture conditions for Robo1-CAR-NK92 cells.

Main Methods:

  • Robo1-CAR-NK92 cells were generated via lentiviral transfection and validated by flow cytometry.
  • Robo1 expression was assessed in U87-MG and SH-SY5Y cell lines.
  • Cytotoxicity was measured using CCK-8 assays and live cell imaging.
  • Cytokine secretion was quantified using multiplex bead-array assays.
  • Cell proliferation, survival, and cytotoxicity were analyzed after treatment with IL-15, IL-21, and dexamethasone.

Main Results:

  • Robo1-CAR-NK92 cells showed significantly higher specific lysis against target cells compared to parental NK92 cells (P<0.05).
  • Robo1-CAR-NK92 cells secreted increased levels of IL-6, IL-10, TNF-α, and IFN-γ during cytotoxic activity.
  • IL-15 enhanced proliferation and survival, while IL-21 had an opposing effect; both enhanced cytotoxicity.
  • Dexamethasone combined with interleukins promoted proliferation and survival but impaired cytotoxicity, except for IL-15 + dexamethasone.

Conclusions:

  • Robo1-CAR-NK92 cells possess enhanced targeted killing capabilities against glioma and neuroblastoma cell lines.
  • The combination of IL-15 and dexamethasone represents an optimized protocol for culturing Robo1 CAR NK cells, promoting proliferation and survival.
Keywords:
GliomaImmunotherapy, adoptiveNeuroblastomaRoundabout protein

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