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Pharmaceutical Research

, Volume 32, Issue 3, pp 779–792 | Cite as

The Narrow-Spectrum HDAC Inhibitor Entinostat Enhances NKG2D Expression Without NK Cell Toxicity, Leading to Enhanced Recognition of Cancer Cells

  • Shiguo ZhuEmail author
  • Cecele J. Denman
  • Zehra S. Cobanoglu
  • Simin Kiany
  • Ching C. Lau
  • Stephen M. Gottschalk
  • Dennis P. M. Hughes
  • Eugenie S. Kleinerman
  • Dean A. LeeEmail author
Research Paper

ABSTRACT

Purpose

Natural killer (NK) cell cytotoxicity correlates with the ligation of activating receptors (e.g., NKG2D) by their ligands (e.g., MHC class I–related chains [MIC] A and B) on target cells. Histone deacetylase inhibitors (HDACi) at high concentrations inhibit tumor growth and can increase NKG2D ligand expression on tumor targets, but are widely regarded as toxic to NK cells.

Methods

We investigated the mechanism of entinostat, a benzamide-derivative narrow-spectrum HDACi, in augmenting the cytotoxicity of NK cells against human colon carcinoma and sarcoma by assessing gene and protein expression, histone acetylation, and cytotoxicity in in vitro and murine models.

Results

We observed that entinostat dose- and time-dependent increase in MIC expression in tumor targets and NKG2D in primary human NK cells, both correlating with increased acetylated histone 3 (AcH3) binding to associated promoters. Entinostat pretreatment of colon carcinoma and sarcoma cells, NK cells, or both led to enhanced overall cytotoxicity in vitro, which was reversed by NKG2D blockade, and inhibited growth of tumor xenografts. Lastly, we showed decreased expression of MICA and ULBP2 transcription in primary human osteosarcoma.

Conclusions

Entinostat enhances NK cell killing of cancer cells through upregulation of both NKG2D and its ligands, suggesting an attractive approach for augmenting NK cell immunotherapy of solid tumors such as colon carcinoma and sarcomas.

KEY WORDS

cancer HDAC inhibitor natural killer cells NKG2D NKG2D ligands 

ABBREVIATIONS

7-AAD

7-Amino-actinomycin D

AcH3

acetylated histone 3

AcH4

acetyl-histone 4

APC

allophycocyanin

ChIP

chromatin immunoprecipitation

FFLuc

firefly luciferase

HDAC

Histone deacetylase

HDACi

Histone deacetylase inhibitors

MIC

MHC class I–related chains

SAHA

suberoylanilide hydroxamic acid

ULBP

UL16-binding proteins

VPA

valproic acid

Notes

ACKNOWLEDGMENTS AND DISCLOSURES

The authors acknowledge Laurence J. N. Cooper for initiating the discussions that led to this work, Joya Chandra for sharing her expertise in epigenetics and critiquing the manuscript, and Shana Palla for guiding our statistical analysis. Financial support for this research was provided to D.A.L. and D.P.M.H. by the Brenda and Howard Johnson Fund, the Physician Scientist Program of MD Anderson Cancer Center, and the Sunbeam Foundation. Financial support to S.Z. was provided by the National Natural Science Foundation of China (81071858; 81273216) and Innovation Program of Shanghai Municipal Education Commission (11ZZ105). STR DNA fingerprinting was done by the Characterized Cell Line Core at MD Anderson Cancer Center, funded by NCI # CA16672. The MSC employed in this work were provided by the Tulane Center for Gene Therapy through a grant from NCRR of the NIH, Grant # P40RR017447.

Supplementary material

11095_2013_1231_Fig8_ESM.jpg (168 kb)
Supplemental Fig. 1

Map indicating the sequence and location of human MICA, MICB and DAP10 promoters, including ATG translation start sites and the promoter-specific forward and reverse PCR primers, relative to the transcription initiation sites (TIS). (JPEG 167 kb)

11095_2013_1231_Fig9_ESM.jpg (41 kb)
Supplemental Fig. 2

Enhanced expression of GFP-Luc transgene in HT1080-GFP-Luc cells treated 24 h with 0.1 – 1 μM entinostat. (JPEG 41 kb)

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Copyright information

© Springer Science+Business Media, LLC (outside the USA) 2013

Authors and Affiliations

  • Shiguo Zhu
    • 1
    Email author
  • Cecele J. Denman
    • 2
  • Zehra S. Cobanoglu
    • 2
  • Simin Kiany
    • 2
  • Ching C. Lau
    • 3
  • Stephen M. Gottschalk
    • 4
  • Dennis P. M. Hughes
    • 2
  • Eugenie S. Kleinerman
    • 2
  • Dean A. Lee
    • 2
    • 5
    Email author
  1. 1.Shanghai University of Traditional Chinese Medicine School of Basic Medical SciencesShanghaiChina
  2. 2.Division of PediatricsThe University of Texas MD Anderson Cancer CenterHoustonUSA
  3. 3.Hematology/Oncology Section, Department of PediatricsBaylor College of MedicineHoustonUSA
  4. 4.Baylor College of Medicine, Department of Pediatrics Center for Cell and Gene TherapyTexas Children’s Cancer CenterHoustonUSA
  5. 5.Pediatrics Unit #853University of Texas MD Anderson Cancer CenterHoustonUSA

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