Chimeric antigen receptor–modified T cells (CAR-T cells) have emerged as a promising cancer immunotherapy for solid tumors. Epithelial cell adhesion molecule (EpCAM) is overexpressed in a variety of tumors and is recognized as a biomarker for circulating tumor cells and cancer stem cells, representing an attractive target for adoptive T-cell immunotherapy. This study generated third-generation CAR-T cells with redirected specificity to EpCAM (EpCAM CAR-T) by lentiviral vector. The study demonstrated that EpCAM CAR-T cells can elicit lytic cytotoxicity to target cells in an EpCAM-dependent manner and secrete cytotoxic cytokines, including interferon gamma and tumor necrosis factor alpha. Furthermore, adoptive transfer of EpCAM CAR-T cells significantly delayed tumor growth and formation in xenograft models. In addition, the safety evaluation showed that CAR-T cells have no systemic toxicity in mice. The data confirmed the antitumor ability and safety of CAR-T cells targeting EpCAM and may provide a new target for CAR-T cell therapies in treating solid tumors.
Get full access to this article
View all access options for this article.
References
1.
HerlynM, SteplewskiZ, HerlynD, et al.Colorectal carcinoma-specific antigen: detection by means of monoclonal antibodies. Proc Natl Acad Sci U S A, 1979; 76:1438–1442.
2.
WentPT, LugliA, MeierS, et al.Frequent EpCam protein expression in human carcinomas. Hum Pathol, 2004; 35:122–128.
3.
NubelT, PreobraschenskiJ, TuncayH, et al.Claudin-7 regulates EpCAM-mediated functions in tumor progression. Mol Cancer Res, 2009; 7:285–299.
4.
BrunnerA, PrelogM, VerdorferI, et al.EpCAM is predominantly expressed in high grade and advanced stage urothelial carcinoma of the bladder. J Clin Pathol, 2008; 61:307–310.
5.
FongD, SteurerM, ObristP, et al.Ep-CAM expression in pancreatic and ampullary carcinomas: frequency and prognostic relevance. J Clin Pathol, 2008; 61:31–35.
6.
SpizzoG, WentP, DirnhoferS, et al.High Ep-CAM expression is associated with poor prognosis in node-positive breast cancer. Breast Cancer Res Treat, 2004; 86:207–213.
7.
VargaM, ObristP, SchneebergerS, et al.Overexpression of epithelial cell adhesion molecule antigen in gallbladder carcinoma is an independent marker for poor survival. Clin Cancer Res, 2004; 10:3131–3136.
8.
JoosseSA, GorgesTM, PantelK. Biology, detection, and clinical implications of circulating tumor cells. EMBO Mol Med, 2015; 7:1–11.
9.
SchulzeK, GaschC, StauferK, et al.Presence of EpCAM-positive circulating tumor cells as biomarker for systemic disease strongly correlates to survival in patients with hepatocellular carcinoma. Int J Cancer, 2013; 133:2165–2171.
10.
WeissensteinU, SchumannA, ReifM, et al.Detection of circulating tumor cells in blood of metastatic breast cancer patients using a combination of cytokeratin and EpCAM antibodies. BMC Cancer, 2012; 12:206.
11.
CoumansFA, DoggenCJ, AttardG, et al.All circulating EpCAM+CK+CD45– objects predict overall survival in castration-resistant prostate cancer. Ann Oncol, 2010; 21:1851–1857.
12.
KhanMS, TsiganiT, RashidM, et al.Circulating tumor cells and EpCAM expression in neuroendocrine tumors. Clin Cancer Res, 2011; 17:337–345.
13.
Al-HajjM, WichaMS, Benito-HernandezA, et al.Prospective identification of tumorigenic breast cancer cells. Proc Natl Acad Sci U S A, 2003; 100:3983–3988.
14.
GiresO, KleinCA, BaeuerlePA. On the abundance of EpCAM on cancer stem cells. Nat Rev Cancer, 2009; 9:143; author reply 143.
15.
LiC, HeidtDG, DalerbaP, et al.Identification of pancreatic cancer stem cells. Cancer Res, 2007; 67:1030–1037.
16.
O'BrienCA, PollettA, GallingerS, et al.A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature, 2007; 445:106–110.
17.
VisvaderJE, LindemanGJ. Cancer stem cells in solid tumours: accumulating evidence and unresolved questions. Nat Rev Cancer, 2008; 8:755–768.
18.
RiethmullerG, HolzE, SchlimokG, et al.Monoclonal antibody therapy for resected Dukes' C colorectal cancer: seven-year outcome of a multicenter randomized trial. J Clin Oncol, 1998; 16:1788–1794.
19.
PuntCJ, NagyA, DouillardJY, et al.Edrecolomab alone or in combination with fluorouracil and folinic acid in the adjuvant treatment of stage III colon cancer: a randomised study. Lancet, 2002; 360:671–677.
20.
FieldsAL, KellerA, SchwartzbergL, et al.Adjuvant therapy with the monoclonal antibody Edrecolomab plus fluorouracil-based therapy does not improve overall survival of patients with stage III colon cancer. J Clin Oncol, 2009; 27:1941–1947.
21.
MarschnerN, RuttingerD, ZugmaierG, et al.Phase II study of the human anti-epithelial cell adhesion molecule antibody adecatumumab in prostate cancer patients with increasing serum levels of prostate-specific antigen after radical prostatectomy. Urol Int, 2010; 85:386–395.
22.
XiangW, WimbergerP, DreierT, et al.Cytotoxic activity of novel human monoclonal antibody MT201 against primary ovarian tumor cells. J Cancer Res Clin Oncol, 2003; 129:341–348.
23.
SchmidtM, RuttingerD, SebastianM, et al.Phase IB study of the EpCAM antibody adecatumumab combined with docetaxel in patients with EpCAM-positive relapsed or refractory advanced-stage breast cancer. Ann Oncol, 2012; 23:2306–2313.
24.
EskanderRN, TewariKS. Epithelial cell-adhesion molecule-directed trifunctional antibody immunotherapy for symptom management of advanced ovarian cancer. Clin Pharmacol, 2013; 5:55–61.
25.
SerranoLM, PfeifferT, OlivaresS, et al.Differentiation of naive cord-blood T cells into CD19-specific cytolytic effectors for posttransplantation adoptive immunotherapy. Blood, 2006; 107:2643–2652.
26.
KowolikCM, ToppMS, GonzalezS, et al.CD28 costimulation provided through a CD19-specific chimeric antigen receptor enhances in vivo persistence and antitumor efficacy of adoptively transferred T cells. Cancer Res, 2006; 66:10995–11004.
27.
MiloneMC, FishJD, CarpenitoC, et al.Chimeric receptors containing CD137 signal transduction domains mediate enhanced survival of T cells and increased antileukemic efficacy in vivo. Mol Ther, 2009; 17:1453–1464.
28.
SongDG, YeQ, CarpenitoC, et al.In vivo persistence, tumor localization, and antitumor activity of CAR-engineered T cells is enhanced by costimulatory signaling through CD137 (4-1BB). Cancer Res, 2011; 71:4617–4627.
29.
SallustoF, LenigD, ForsterR, et al.Two subsets of memory T lymphocytes with distinct homing potentials and effector functions. Nature, 1999; 401:708–712.
30.
SallustoF, GeginatJ, LanzavecchiaA. Central memory and effector memory T cell subsets: function, generation, and maintenance. Annu Rev Immunol, 2004; 22:745–763.
CarpenitoC, MiloneMC, HassanR, et al.Control of large, established tumor xenografts with genetically retargeted human T cells containing CD28 and CD137 domains. Proc Natl Acad Sci U S A, 2009; 106:3360–3365.
33.
ZhongXS, MatsushitaM, PlotkinJ, et al.Chimeric antigen receptors combining 4-1BB and CD28 signaling domains augment PI3kinase/AKT/Bcl-XL activation and CD8+ T cell-mediated tumor eradication. Mol Ther, 2010; 18:413–420.
34.
TillBG, JensenMC, WangJ, et al.CD20-specific adoptive immunotherapy for lymphoma using a chimeric antigen receptor with both CD28 and 4-1BB domains: pilot clinical trial results. Blood, 2012; 119:3940–3950.
35.
TammanaS, HuangX, WongM, et al.4-1BB and CD28 signaling plays a synergistic role in redirecting umbilical cord blood T cells against B-cell malignancies. Hum Gene Ther, 2010; 21:75–86.
36.
KarlssonH, SvenssonE, GiggC, et al.Evaluation of intracellular signaling downstream chimeric antigen receptors. PLoS One, 2015; 10:e0144787.
37.
HeczeyA, LiuD, TianG, et al.Invariant NKT cells with chimeric antigen receptor provide a novel platform for safe and effective cancer immunotherapy. Blood, 2014; 124:2824–2833.
38.
YangS, GattinoniL, LiuF, et al.In vitro generated anti-tumor T lymphocytes exhibit distinct subsets mimicking in vivo antigen-experienced cells. Cancer Immunol Immunother, 2011; 60:739–749.
39.
KlebanoffCA, GattinoniL, Torabi-PariziP, et al.Central memory self/tumor-reactive CD8+ T cells confer superior antitumor immunity compared with effector memory T cells. Proc Natl Acad Sci U S A, 2005; 102:9571–9576.
40.
BergerC, JensenMC, LansdorpPM, et al.Adoptive transfer of effector CD8+ T cells derived from central memory cells establishes persistent T cell memory in primates. J Clin Invest, 2008; 118:294–305.
41.
DengZ, WuY, MaW, et al.Adoptive T-cell therapy of prostate cancer targeting the cancer stem cell antigen EpCAM. BMC Immunol, 2015; 16:1.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
