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Systemic delivery of chTNT-3/CpG immunoconjugates for immunotherapy in murine solid tumor models

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Abstract

CpG oligodeoxynucleotides (CpG) potently activate the immune system by mimicking microbial DNA. Conjugation of CpG to chTNT-3, an antibody targeting the necrotic centers of tumors, enabled CpG to accumulate in tumors after systemic delivery, where it can activate the immune system in the presence of tumor antigens. CpG chemically conjugated to chTNT-3 (chTNT-3/CpG) were compared to free CpG in their ability to stimulate the immune system in vitro and reduce tumor burden in vivo. In subcutaneous Colon 26 adenocarcinoma and B16-F10 melanoma models in BALB/c and C57BL/6 mice, respectively, chTNT-3/CpG, free CpG, or several different control constructs were administered systemically. Intraperitoneal injections of chTNT-3/CpG delayed tumor growth and improved survival and were comparable to intratumorally administered CpG. Compared to saline-treated mice, chTNT-3/CpG-treated mice had smaller average tumor volumes by as much as 72 % in Colon 26-bearing mice and 79 % in B16-bearing mice. Systemically delivered free CpG and CpG conjugated to an isotype control antibody did not reduce tumor burden or improve survival. In this study, chTNT-3/CpG retained immunostimulatory activity of the CpG moiety and enabled delivery to tumors. Because systemically administered CpG rapidly clear the body and do not accumulate into tumors, chTNT-3/CpG provide a solution to the limitations observed in preclinical and clinical trials.

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Abbreviations

ANOVA:

Analysis of variance

cetuximab/CpG:

CpG conjugated to cetuximab

chTNT-3:

Chimeric tumor necrosis therapy-3 antibody

chTNT-3/CpG:

CpG conjugated to chTNT-3

chTNT-3+CpG:

Non-conjugated chTNT-3 and CpG

CpG:

Cytosine guanine dinucleotide

CpG1585:

CpG sequence 1585

CpG1826:

CpG sequence 1826

DC:

Dendritic cell

DNP:

Dinitrophenyl haptens

EGFR:

Epidermal growth factor receptor

GEE:

Generalized estimating equations

125I-chTNT-3/CpG:

125-Iodinated chTNT-3/CpG

IFN:

Interferon

i.p.:

Intraperitoneal

i.t.:

Intratumoral

NK cells:

Natural killer cells

PAMPs:

Pathogen-associated molecular patterns

sc1585:

Negative control sequence for CpG1585

sc1826:

Negative control sequence for CpG1826

TCR:

T cell receptors

TDLN:

Tumor-draining lymph node

TH1:

T-helper 1

TLR:

Toll-like receptors

Treg:

T regulatory cells

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Acknowledgments

The authors would like to acknowledge Dr. Wendy Jean Mack (University of Southern California) for statistical assistance. The project described was supported in part by National Center for Advancing Translational Sciences (NCATS award UL1TR00013) and National Cancer Institute (NCI award P30CA014089). Julie K. Jang and Brian W. Wu are TL1 trainees under Southern California Clinical and Translational Science Institute (SC CTSI) (NCATS award TL1TR000132).

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Authors and Affiliations

  1. Department of Pathology, Keck School of Medicine, University of Southern California, 2011 Zonal Avenue, HMR 205, Los Angeles, CA, 90033, USA

    Julie K. Jang, Leslie A. Khawli, David C. Canter, Peisheng Hu, Tian H. Zhu, Brian W. Wu & Alan L. Epstein

  2. Department of Endocrinology, Metabolism, and Hypertension, Thyroid Section, Brigham and Women’s Hospital, Boston, MA, USA

    Trevor E. Angell

  3. Department of Blood Transfusion, Xinqiao Hospital, Third Military Medical University, Chongqing, China

    Zhongjun Li

Authors
  1. Julie K. Jang
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  2. Leslie A. Khawli
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  3. David C. Canter
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  4. Peisheng Hu
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  5. Tian H. Zhu
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  7. Trevor E. Angell
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  8. Zhongjun Li
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  9. Alan L. Epstein
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Correspondence to Alan L. Epstein.

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Conflict of interest

Peisheng Hu and Alan L. Epstein are co-founders of Cancer Therapeutics Laboratories, Inc. This study was partially funded by Cancer Therapeutics Laboratories, Inc. All other authors declare no conflict of interest.

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Jang, J.K., Khawli, L.A., Canter, D.C. et al. Systemic delivery of chTNT-3/CpG immunoconjugates for immunotherapy in murine solid tumor models. Cancer Immunol Immunother 65, 511–523 (2016). https://doi.org/10.1007/s00262-016-1813-x

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