Abstract
The envelope (env) protein of the human endogenous retrovirus type K (HERV-K) family is commonly expressed on the surface of breast cancer cells. We assessed whether HERV-K env is a potential target for antibody-based immunotherapy of breast cancer. We examined the expression of HERV-K env protein in various malignant (MDA-MB-231, MCF-7, SKBR3, MDA-MB-453, T47D, and ZR-75-1) and nonmalignant (MCF-10A and MCF-10AT) human breast cell lines by immunoblot, enzyme-linked immunosorbent assay, immunofluorescence staining, and flow cytometry. Anti-HERV-K env monoclonal antibodies (mAbs; 6H5, 4D1, 4E11, 6E11, and 4E6) were used to target expression of HERV-K, and antitumor effects were assessed by quantifying growth and apoptosis of breast cancer cells in vitro, and tumor growth in vivo in mice (n = 5 per group) bearing xenograft tumors. The mechanisms responsible for 6H5 mAb-mediated effects were investigated by microarray assays, flow cytometry, immunoblot, and immunofluorescence staining. The expression of HERV-K env protein was assessed in primary breast tumors (n = 223) by immunohistochemistry. All statistical tests were two-sided. The expression of HERV-K env protein in malignant breast cancer cell lines was substantially higher than nonmalignant breast cells. Anti-HERV-K-specific mAbs inhibited growth and induced apoptosis of breast cancer cells in vitro. Mice treated with 6H5 mAb showed statistically significantly reduced growth of xenograft tumors compared with mice treated with control immunoglobulin (control [mIgG] vs 6H5 mAb, for tumors originating from MDA-MB-231 cells, mean size = 1448.33 vs 475.44 mm(3); difference = 972.89 mm(3), 95% CI = 470.17 to 1475.61 mm(3); P < .001). Several proteins involved in the apoptotic signaling pathways were overexpressed in vitro in 6H5 mAb-treated malignant breast cells compared with mIgG-treated control. HERV-K expression was detected in 148 (66%) of 223 primary breast tumors, and a higher rate of lymph node metastasis was associated with HERV-K-positive compared with HERV-K-negative tumors (43% vs 23%, P = .003). Monoclonal antibodies against HERV-K env protein show potential as novel immunotherapeutic agents for breast cancer therapy.
MeSH Terms
Adult
Aged
Animals
Antibodies, Anti-Idiotypic/pharmacology
Antibodies, Monoclonal/pharmacology
Antineoplastic Agents/pharmacology
Apoptosis/drug effects
Breast Neoplasms/drug therapy,immunology,pathology,virology
Bromodeoxyuridine/metabolism
Carcinoma, Ductal, Breast/drug therapy,virology
Cell Line, Tumor
Endogenous Retroviruses
Enzyme-Linked Immunosorbent Assay
Feasibility Studies
Female
Flow Cytometry
Fluorescent Antibody Technique
Gene Expression Regulation, Neoplastic
Gene Expression Regulation, Viral
Gene Products, env/antagonists & inhibitors,metabolism
Humans
Immunoblotting
Immunohistochemistry
Immunotherapy/methods
In Situ Nick-End Labeling
Ki-67 Antigen/analysis
Mice
Mice, Nude
Middle Aged
Molecular Targeted Therapy
Neoplasm Grading
Neoplasm Staging
Pilot Projects
Protein Array Analysis
Random Allocation
Retroviridae Proteins/antagonists & inhibitors,metabolism
Signal Transduction
Transplantation, Heterologous
Tumor Suppressor Protein p53/metabolism
Up-Regulation
Chemicals
Antibodies, Anti-Idiotypic
Antibodies, Monoclonal
Antineoplastic Agents
Gene Products, env
Ki-67 Antigen
Retroviridae Proteins
TP53 protein, human
Tumor Suppressor Protein p53
Bromodeoxyuridine
Authors & Affiliations
15 authors, click to expand affiliations / ORCID
Wang-Johanning Feng
Department of Veterinary Sciences, The University of Texas MD Anderson Cancer Center, Houston, TX, USA. fwangjoh@mdanderson.org
Rycaj Kiera
Plummer Joshua B
Li Ming
Yin Bingnan
Frerich Katherine
Garza Jeremy G
Shen Jianjun
Lin Kevin
Yan Peisha
Glynn Sharon A
Dorsey Tiffany H
Hunt Kelly K
Ambs Stefan
Johanning Gary L
References (38)
38 references, click to expand
-
Stabilised cellular immuno-fluorescence assay: CD45 expression as a calibration standard for human leukocytes.
J Immunol Methods. 2002 Aug 1;266(1-2):19-32
PMID: 12133619
-
Expression patterns of transcribed human endogenous retrovirus HERV-K(HML-2) loci in human tissues and the need for a HERV Transcriptome Project.
BMC Genomics. 2008 Jul 29;9:354
PMID: 18664271
-
Response to therapy in breast cancer.
J Nucl Med. 2009 May;50 Suppl 1:55S-63S
PMID: 19380410
-
Human endogenous retrovirus (HERV-K) reverse transcriptase as a breast cancer prognostic marker.
Neoplasia. 2008 Jun;10(6):521-33
PMID: 18516289
-
Type 1 HERV-K genome is spliced into subgenomic transcripts in the human breast tumor cell line T47D.
Virology. 1997 Aug 4;234(2):304-8
PMID: 9268162
-
Expression of multiple human endogenous retrovirus surface envelope proteins in ovarian cancer.
Int J Cancer. 2007 Jan 1;120(1):81-90
PMID: 17013901
-
Molecular cloning and long terminal repeat sequences of human endogenous retrovirus genes related to types A and B retrovirus genes.
J Virol. 1986 Jun;58(3):937-44
PMID: 3009897
-
Design, expression, purification, and characterization, in vitro and in vivo, of an antimelanoma single-chain Fv antibody fused to the toxin gelonin.
Cancer Res. 2003 Jul 15;63(14):3995-4002
PMID: 12873997
-
The Nottingham Prognostic Index in primary breast cancer.
Breast Cancer Res Treat. 1992;22(3):207-19
PMID: 1391987
-
Initial sequencing and analysis of the human genome.
Nature. 2001 Feb 15;409(6822):860-921
PMID: 11237011
-
Increased NOS2 predicts poor survival in estrogen receptor-negative breast cancer patients.
J Clin Invest. 2010 Nov;120(11):3843-54
PMID: 20978357
-
Human endogenous retrovirus HERV-K(HML-2) encodes a stable signal peptide with biological properties distinct from Rec.
Retrovirology. 2009 Feb 16;6:17
PMID: 19220907
-
Monoclonal antibody therapeutics and apoptosis.
Oncogene. 2003 Dec 8;22(56):9097-106
PMID: 14663488
-
Identification of CXCR4 as a new nitric oxide-regulated gene in human CD34+ cells.
Stem Cells. 2007 Jan;25(1):211-9
PMID: 17023512
-
Kinetics of anti-carcinoembryonic antigen antibody internalization: effects of affinity, bivalency, and stability.
Cancer Immunol Immunother. 2008 Dec;57(12):1879-90
PMID: 18408925
-
Stimulation of expression of the human endogenous retrovirus genome by female steroid hormones in human breast cancer cell line T47D.
J Virol. 1987 Jun;61(6):2059-62
PMID: 2883329
-
Endogenous retroviruses in systemic response to stress signals.
Shock. 2008 Aug;30(2):105-16
PMID: 18317406
-
Cdk5 phosphorylates non-genotoxically overexpressed p53 following inhibition of PP2A to induce cell cycle arrest/apoptosis and inhibits tumor progression.
Mol Cancer. 2010 Jul 31;9:204
PMID: 20673369
-
An almost-intact human endogenous retrovirus K on human chromosome 7.
Nat Genet. 1999 Mar;21(3):257-8
PMID: 10080172
-
A balance between NF-Y and p53 governs the pro- and anti-apoptotic transcriptional response.
Nucleic Acids Res. 2008 Mar;36(5):1415-28
PMID: 18187512
-
Retrovirus-like particles released from the human breast cancer cell line T47-D display type B- and C-related endogenous retroviral sequences.
J Virol. 1995 Oct;69(10):6408-16
PMID: 7545247
-
Expression of human endogenous retrovirus K in melanomas and melanoma cell lines.
Cancer Res. 2005 May 15;65(10):4172-80
PMID: 15899808
-
Transcriptional control of human p53-regulated genes.
Nat Rev Mol Cell Biol. 2008 May;9(5):402-12
PMID: 18431400
-
Inflammation and IGF-I activate the Akt pathway in breast cancer.
Int J Cancer. 2007 Feb 15;120(4):796-805
PMID: 17096325
-
Human endogenous retrovirus K (HML-2) elements in the plasma of people with lymphoma and breast cancer.
J Virol. 2008 Oct;82(19):9329-36
PMID: 18632860
-
TNM Classification of Malignant Tumors, fifth edition (1997). Union Internationale Contre le Cancer and the American Joint Committee on Cancer.
Cancer. 1997 Nov 1;80(9):1803-4
PMID: 9351551
-
Expression of the human endogenous retrovirus HTDV/HERV-K is enhanced by cellular transcription factor YY1.
J Virol. 1999 Feb;73(2):1254-61
PMID: 9882329
-
HERV-K(HML-2) GAG/ENV antibodies as indicator for therapy effect in patients with germ cell tumors.
Int J Cancer. 2004 Jun 20;110(3):459-61
PMID: 15095315
-
Quantitation of HERV-K env gene expression and splicing in human breast cancer.
Oncogene. 2003 Mar 13;22(10):1528-35
PMID: 12629516
-
Serological response to human endogenous retrovirus K in melanoma patients correlates with survival probability.
AIDS Res Hum Retroviruses. 2008 May;24(5):717-23
PMID: 18462078
-
Why do viruses cause cancer? Highlights of the first century of human tumour virology.
Nat Rev Cancer. 2010 Dec;10(12):878-89
PMID: 21102637
-
Expression of human endogenous retrovirus k envelope transcripts in human breast cancer.
Clin Cancer Res. 2001 Jun;7(6):1553-60
PMID: 11410490
-
The American Joint Committee on Cancer: the 7th edition of the AJCC cancer staging manual and the future of TNM.
Ann Surg Oncol. 2010 Jun;17(6):1471-4
PMID: 20180029
-
Intracellular expression of a single-chain antibody directed against human papillomavirus type 16 E7 oncoprotein achieves targeted antineoplastic effects.
Cancer Res. 1998 May 1;58(9):1893-900
PMID: 9581830
-
Potential mechanisms of endogenous retroviral-mediated genomic instability in human cancer.
Semin Cancer Biol. 2010 Aug;20(4):246-53
PMID: 20685251
-
A novel multiplex RT-PCR system detects human endogenous retrovirus-K in breast cancer.
Arch Virol. 2005 Jan;150(1):177-84
PMID: 15449135
-
Human endogenous retrovirus K triggers an antigen-specific immune response in breast cancer patients.
Cancer Res. 2008 Jul 15;68(14):5869-77
PMID: 18632641
-
Nitric oxide and oxygen radicals in infection, inflammation, and cancer.
Biochemistry (Mosc). 1998 Jul;63(7):854-65
PMID: 9721338