MP83-04 SUPPRESSION OF HUMAN PROSTATE CANCER (CAP) BY ADOPTIVE TRANSFER OF PATIENT-DERIVED PSMA-SPECIFIC, TGF-ß-INSENSITIVE CD8+ T CELLS

Abstract

You have accessJournal of UrologyProstate Cancer: Basic Research & Pathophysiology II1 Apr 2017MP83-04 SUPPRESSION OF HUMAN PROSTATE CANCER (CAP) BY ADOPTIVE TRANSFER OF PATIENT-DERIVED PSMA-SPECIFIC, TGF-ß-INSENSITIVE CD8+ T CELLS Qiang Zhang, Timothy Kuzel, Brian Helfand, Ximing J Yang, Weijun Qin, Chung Lee, Benebito Carneiro, and Francis J Giles Qiang ZhangQiang Zhang More articles by this author , Timothy KuzelTimothy Kuzel More articles by this author , Brian HelfandBrian Helfand More articles by this author , Ximing J YangXiming J Yang More articles by this author , Weijun QinWeijun Qin More articles by this author , Chung LeeChung Lee More articles by this author , Benebito CarneiroBenebito Carneiro More articles by this author , and Francis J GilesFrancis J Giles More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.2572AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES It has previously been shown that tumor-derived TGF-β related potent immunosuppression and lack of tumor killing specificity promote CaP progression. Here we report a new immunotherapeutic approach using adoptive transfer of patient-derived PSMA-specific, TGF-β-insensitive human CD8+ T cells to inhibit solid xenograft CaP. METHODS Peripheral blood CD8+ T cells were collected from metastatic castration resistant CaP patient by leukapheresis and cultured in the FDA approved Cell Processing Work Station (CPWS, Panasonic) with CD-3 Biotin/CD28/Anti-Biotin Beads (Bead: Cell 1:2) and IL-2 (100units/ml). We developed a TβRIIDN-TK-IRES-PZ1 chimeric T cell receptor retroviral construct using an anti-PSMA IgTCR(ζ) gene (PZ1) and a dominant negative TGF-β type II receptor (TβRIIDN) that could induce CD8+ T cells to be PSMA reactive and insensitive to TGF-β. PC-3 cells (PSMA negative; ATCC) or PSMA positive PC-3-PSMA cells (Cleveland Clinic) were used for target cells. Subcutaneous injection of PC-3 and PC-3-PSMA cells (2x105 cells/each, infected with HSV1-tk-GFP-luciferase reporter) into the left and right flank region respectively in each of 48 immunodeficient RAG-1 mice was performed. One week later, the animals were randomly assigned to one of three adoptive transfer groups (16 mice /each group, 2x106 CD8+ T cells/each mice): Group 1: CD8+ T cells infected with TβRIIDN-TK-IRES-PZ1 (71.1% positive); Group 2: Naive CD8+ T cells; Group 3: No treatment group. Tumor growth was monitored by IVIS luciferase imaging. The animals were provided 2 weekly adoptive transfer treatments and sacrificed after 3 weeks. The size and the weight of the tumor were recorded, and the infiltration of CD8+ T cells and apoptosis was evaluated by immunofluorescence staining and TUNEL. RESULTS In Group 1, the average tumor weight and volume was significantly lower in the PC3-PSMA tumor (0.413g and 514.2mm3) compared to the PC3 tumor (2.75 g and 3165.2 mm3); (~3-6×, P<0.05). There was no difference between the PC3 tumor (2.36g and 2768.5mm3 ) compared to PC3-PSMA tumor (2.45g and 2411.99 mm3) in Group 2 or Group 3. H&E staining showed large amount of nuclear fusion, fragmentation and necrosis were found in PC3-PSMA tumors in Group 1 compared to Group 2 and 3. In Group 1, tumor apoptosis (72.5/1,000 µm2) and CD8+ T cell infiltration (45.5/1,000 µm2 ) in PC3-PSMA tumor parenchyma was significantly higher compared to PC3 tumor (6.7/1,000 µm2 and 3.1/1,000 µm2 respectively). There was no significant apoptosis or CD8+ T cells infiltration observed in either PC3 and PC3-PSMA tumor in Group 2 and Group 3. CONCLUSIONS Our approach combines TGF-β insensitive with PSMA selectivity that significantly enhance the specificity and tumor killing ability of patient CD8+ T cells, and simultaneously suppress the tumor derived TGF-βinduced immunosuppression. Therefore, this construct may offer a novel therapeutic intervention for both primary CaP treatment as well as for recurrence after prostatectomy. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e1107 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Qiang Zhang More articles by this author Timothy Kuzel More articles by this author Brian Helfand More articles by this author Ximing J Yang More articles by this author Weijun Qin More articles by this author Chung Lee More articles by this author Benebito Carneiro More articles by this author Francis J Giles More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...