Despite advances in the understanding of its molecular pathophysiology, pancreatic cancer remains largely incurable, highlighting the need for novel therapies. We developed a chimeric antigen receptor (CAR) specific for prostate stem cell antigen (PSCA), a glycoprotein that is overexpressed in pancreatic cancer starting at early stages of malignant transformation. To optimize the CAR design, we used antigen-recognition domains derived from mouse or human antibodies, and intracellular signaling domains containing one or two T cell costimulatory elements, in addition to CD3zeta. Comparing multiple constructs established that the CAR based on human monoclonal antibody Ha1-4.117 had the greatest reactivity in vitro. To further analyze this CAR, we developed a human pancreatic cancer xenograft model and adoptively transferred CAR-engineered T cells into animals with established tumors. CAR-engineered human lymphocytes induced significant antitumor activity, and unlike what has been described for other CARs, a second-generation CAR (containing CD28 cosignaling domain) induced a more potent antitumor effect than a third-generation CAR (containing CD28 and 41BB cosignaling domains). While our results provide evidence to support PSCA as a target antigen for CAR-based immunotherapy of pancreatic cancer, the expression of PSCA on selected normal tissues could be a source of limiting toxicity.
Get full access to this article
View all access options for this article.
References
1.
AnurathapanU., ChanR.C., HindiH.F., et al. (2014). Kinetics of tumor destruction by chimeric antigen receptor-modified T cells. Mol. Ther., 22, 623–633.
2.
ArganiP., RostyC., ReiterR.E., et al. (2001). Discovery of new markers of cancer through serial analysis of gene expression: prostate stem cell antigen is overexpressed in pancreatic adenocarcinoma. Cancer Res., 61, 4320–4324.
3.
BahrenbergG., BrauersA., JoostH.G., and JakseG. (2000). Reduced expression of PSCA, a member of the LY-6 family of cell surface antigens, in bladder, esophagus, and stomach tumors. Biochem. Biophys. Res. Commun., 275, 783–788.
4.
BrentjensR., YehR., BernalY., et al. (2010). Treatment of chronic lymphocytic leukemia with genetically targeted autologous T cells: case report of an unforeseen adverse event in a phase I clinical trial. Mol. Ther., 18, 666–668.
5.
CarpenitoC., MiloneM.C., HassanR., et al. (2009). Control of large, established tumor xenografts with genetically retargeted human T cells containing CD28 and CD137 domains. Proc. Natl. Acad. Sci. USA, 106, 3360–3365.
6.
CartwrightT., RichardsD.A., and BoehmK.A. (2008). Cancer of the pancreas: are we making progress? A review of studies in the US Oncology Research Network. Cancer Control, 15, 308–313.
7.
ChmielewskiM., HahnO., RapplG., et al. (2012). T cells that target carcinoembryonic antigen eradicate orthotopic pancreatic carcinomas without inducing autoimmune colitis in mice. Gastroenterology, 143, 1095–1107.e2.
8.
Di StasiA., TeyS.K., DottiG., et al. (2011). Inducible apoptosis as a safety switch for adoptive cell therapy. N. Engl. J. Med., 365, 1673–1683.
9.
ElsammanE., FukumoriT., KasaiT., et al. (2006). Prostate stem cell antigen predicts tumour recurrence in superficial transitional cell carcinoma of the urinary bladder. BJU Int., 97, 1202–1207.
10.
FeldmannG., BeatyR., HrubanR.H., and MaitraA. (2007). Molecular genetics of pancreatic intraepithelial neoplasia. J. Hepatobiliary Pancreat. Surg., 14, 224–232.
11.
GattinoniL., LugliE., JiY., et al. (2011). A human memory T cell subset with stem cell-like properties. Nat Med., 17, 1290–1297.
12.
GuZ., ThomasG., YamashiroJ., et al. (2000). Prostate stem cell antigen (PSCA) expression increases with high gleason score, advanced stage and bone metastasis in prostate cancer. Oncogene, 19, 1288–1296.
13.
GudasJ., JakobovitsA., JiaX., et al. (2012). Antibodies and Related Molecules That Bind to PSCA Proteins (Agensys, Inc., Santa Monica, CA).
14.
HillerdalV., RamachandranM., LejaJ., and EssandM. (2014). Systemic treatment with CAR-engineered T cells against PSCA delays subcutaneous tumor growth and prolongs survival of mice. BMC Cancer, 14, 30.
KatariU.L., KeirnanJ.M., WorthA.C., et al. (2011). Engineered T cells for pancreatic cancer treatment. HPB (Oxford), 13, 643–650.
17.
KershawM.H., WestwoodJ.A., ParkerL.L., et al. (2006). A phase I study on adoptive immunotherapy using gene-modified T cells for ovarian cancer. Clin. Cancer Res., 12, 6106–6115.
18.
KlossC.C., CondominesM., CartellieriM., et al. (2013). Combinatorial antigen recognition with balanced signaling promotes selective tumor eradication by engineered T cells. Nat Biotechnol., 31, 71–75.
19.
KochenderferJ.N., DudleyM.E., FeldmanS.A., et al. (2012). B-cell depletion and remissions of malignancy along with cytokine-associated toxicity in a clinical trial of anti-CD19 chimeric-antigen-receptor-transduced T cells. Blood, 119, 2709–2720.
20.
KoidoS., HommaS., TakaharaA., et al. (2011). Current immunotherapeutic approaches in pancreatic cancer. Clin. Dev. Immunol., 2011, 267539.
21.
LamersC.H., SleijferS., VultoA.G., et al. (2006). Treatment of metastatic renal cell carcinoma with autologous T-lymphocytes genetically retargeted against carbonic anhydrase IX: first clinical experience. J. Clin. Oncol., 24, e20–e22.
22.
LamersC.H., WillemsenR., Van ElzakkerP., et al. (2011). Immune responses to transgene and retroviral vector in patients treated with ex vivo-engineered T cells. Blood, 117, 72–82.
23.
LeytonJ.V., OlafsenT., LepinE.J., et al. (2008). Humanized radioiodinated minibody for imaging of prostate stem cell antigen-expressing tumors. Clin. Cancer Res., 14, 7488–7496.
24.
LeytonJ.V., OlafsenT., ShermanM.A., et al. (2009). Engineered humanized diabodies for microPET imaging of prostate stem cell antigen-expressing tumors. Protein Eng. Des. Sel., 22, 209–216.
25.
LiQ., VerschraegenC.F., MendozaJ., and HassanR. (2004). Cytotoxic activity of the recombinant anti-mesothelin immunotoxin, SS1(dsFv)PE38, towards tumor cell lines established from ascites of patients with peritoneal mesotheliomas. Anticancer Res., 24, 1327–1335.
26.
MaliarA., ServaisC., WaksT., et al. (2012). Redirected T cells that target pancreatic adenocarcinoma antigens eliminate tumors and metastases in mice. Gastroenterology, 143, 1375–1384.e1–e5.
27.
MausM.V., HaasA.R., BeattyG.L., et al. (2013). T cells expressing chimeric antigen receptors can cause anaphylaxis in humans. Cancer Immunol. Res., 1, 26–31.
28.
MorganR.A., YangJ.C., KitanoM., et al. (2010). Case report of a serious adverse event following the administration of T cells transduced with a chimeric antigen receptor recognizing ERBB2. Mol. Ther., 18, 843–851.
29.
MorganR.A., ChinnasamyN., Abate-DagaD., et al. (2013). Cancer regression and neurological toxicity following anti-MAGE-A3 TCR gene therapy. J. Immunother., 36, 133–151.
30.
MorgenrothA., CartellieriM., SchmitzM., et al. (2007). Targeting of tumor cells expressing the prostate stem cell antigen (PSCA) using genetically engineered T-cells. Prostate, 67, 1121–1131.
31.
OnoH., YanagiharaK., SakamotoH., et al. (2012). Prostate stem cell antigen gene is expressed in islets of pancreas. Anat. Cell Biol., 45, 149–154.
32.
OstermannE., Garin-ChesaP., HeiderK.H., et al. (2008). Effective immunoconjugate therapy in cancer models targeting a serine protease of tumor fibroblasts. Clin. Cancer Res., 14, 4584–4592.
33.
RobbinsP.F., MorganR.A., FeldmanS.A., et al. (2011). Tumor regression in patients with metastatic synovial cell sarcoma and melanoma using genetically engineered lymphocytes reactive with NY-ESO-1. J. Clin. Oncol., 29, 917–924.
34.
SaekiN., GuJ., YoshidaT., and WuX. (2010). Prostate stem cell antigen: a Jekyll and Hyde molecule?. Clin. Cancer Res., 16, 3533–3538.
35.
SiegelR., NaishadhamD., and JemalA. (2012). Cancer statistics, 2012. CA Cancer J. Clin., 62, 10–29.
36.
TammanaS., HuangX., WongM., et al. (2010). 4-1BB and CD28 signaling plays a synergistic role in redirecting umbilical cord blood T cells against B-cell malignancies. Hum. Gene Ther., 21, 75–86.
37.
WolpinB.M., O'ReillyE.M., KoY.J., et al. (2013). Global, multicenter, randomized, phase II trial of gemcitabine and gemcitabine plus AGS-1C4D4 in patients with previously untreated, metastatic pancreatic cancer. Ann. Oncol., 24, 1792–1801.
38.
ZhaoY., WangQ.J., YangS., et al. (2009). A herceptin-based chimeric antigen receptor with modified signaling domains leads to enhanced survival of transduced T lymphocytes and antitumor activity. J. Immunol., 183, 5563–5574.
39.
ZhengZ., ChinnasamyN., and MorganR.A. (2012). Protein L: a novel reagent for the detection of chimeric antigen receptor (CAR) expression by flow cytometry. J. Transl. Med., 10, 29.
40.
ZhongX.S., MatsushitaM., PlotkinJ., et al. (2010). 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., 18, 413–420.
41.
ZhouX., LiJ., WangZ., et al. (2013). Cellular immunotherapy for carcinoma using genetically modified EGFR-specific T lymphocytes. Neoplasia, 15, 544–553.
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.
