iRGD-modified memory-like NK cells exhibit potent responses to hepatocellular carcinoma

Yanbing Dong; Ying Huang; Zhe Zhang; Aoxing Chen; Lin Li; Manman Tian; Jie Shen; Jie Shao

doi:10.1186/s12967-023-04024-7

iRGD-modified memory-like NK cells exhibit potent responses to hepatocellular carcinoma

J Transl Med. 2023 Mar 17;21(1):205. doi: 10.1186/s12967-023-04024-7.

Authors

Yanbing Dong^#¹, Ying Huang^#^{2

3}, Zhe Zhang², Aoxing Chen², Lin Li², Manman Tian¹, Jie Shen⁴, Jie Shao^{5

6}

Affiliations

¹ Comprehensive Cancer Centre of Drum Tower Hospital, Medical School of Nanjing University, Clinical Cancer Institute of Nanjing University, Nanjing, China.
² Comprehensive Cancer Center, Nanjing Drum Tower Hospital Clinical College of Traditional Chinese and Western Medicine, Nanjing University of Traditional Chinese Medicine, Nanjing, China.
³ Department of Oncology, The Second People's Hospital of Huai'an, Huai'an, China.
⁴ Comprehensive Cancer Centre of Drum Tower Hospital, Medical School of Nanjing University, Clinical Cancer Institute of Nanjing University, Nanjing, China. shenjie2008nju@163.com.
⁵ Comprehensive Cancer Centre of Drum Tower Hospital, Medical School of Nanjing University, Clinical Cancer Institute of Nanjing University, Nanjing, China. sj830616@163.com.
⁶ Comprehensive Cancer Center, Nanjing Drum Tower Hospital Clinical College of Traditional Chinese and Western Medicine, Nanjing University of Traditional Chinese Medicine, Nanjing, China. sj830616@163.com.

^# Contributed equally.

Abstract

Background: Cytokine-induced memory-like natural killer (CIML NK) cells have been found to possess potent antitumor responses and induce complete remissions in patients with leukemia. However, the poor infiltration of transferred NK cells is a major obstacle in developing adoptive cell immunotherapy for solid tumors. In our study, we explored the potential of using the tumor-penetrating peptide iRGD to deliver activated CIML NK cells deep into tumor tissues.

Methods: After being briefly stimulated with interleukin-12 (IL-12), IL-15, and IL-18, CIML NK cells were assessed for their phenotype and function with flow cytometry. The penetrating and killing capability of iRGD-modified CIML NK cells in tumor spheroids was revealed by confocal microscopy. The anti-tumor efficacy of these modified CIML NK cells was tested in hepatocellular carcinoma (HCC) xenograft mouse models.

Results: Treating NK cells with cytokines led to a substantial activation, which was evidenced by the upregulation of CD25 and CD137. After a resting period of six days, CIML NK cells were still able to display strong activation when targeting HepG2 and SK-Hep-1 HCC cell lines. Additionally, CIML NK cells produced increased amounts of cytokines (interferon-gamma and tumor necrosis factor alpha) and exhibited heightened cytotoxicity towards HCC cell lines. The iRGD modification enabled CIML NK cells to infiltrate multicellular spheroids (MCSs) and, consequently, to induce cytotoxicity against the target cancer cells. Moreover, the CIML NK cells modified with iRGD significantly decreased tumor growth in a HCC xenograft mouse model.

Conclusion: Our findings demonstrate that CIML NK cells possess augmented potency and durability against HCC cell lines in vitro. Additionally, we have seen that the incorporation of iRGD to CIML NK cells facilitates enhanced infiltration and targeted destruction of MCSs. Moreover, the application of iRGD-modified CIML NK cells reveal remarkable anti-tumor efficacy against HCC in vivo.

Keywords: Hepatocellular carcinoma; Memory-like NK cells; iRGD.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Carcinoma, Hepatocellular* / pathology
Carcinoma, Hepatocellular* / therapy
Cell Line, Tumor
Cytokines / metabolism
Humans
Interleukin-12 / pharmacology
Killer Cells, Natural / metabolism
Liver Neoplasms* / therapy
Mice

Substances

Cytokines
Interleukin-12