292 Final Results of Phase I Trial of GMCSF Gene TGFß2 Antisense Gene Autologous Tumor Cell (TAG) Vaccine in Advanced Cancer Molecular Therapy Volume 18, Supplement 1, May 2010 Copyright © The America[.]
CLINICAL GENE & CELL THERAPY ORAL ABSTRACT SESSION at months To evaluate whether long term gene expression was achievable, a second cohort of patients was evaluated at months As in the rst cohort, a double-blind, randomized controlled trial was initiated in LGMD2D subjects using rAAV1 with full length human SGCA under control of a truncated muscle creatine kinase (tMCK) promoter The extensor digitorum brevis (EDB) muscle received 3.25 X 1011 vector genomes on one side, while the control side was given saline All analyses were done prior to breaking the blind The entire EDB muscle was removed from both sides at months in all three subjects Transgene specic PCR analysis demonstrated expression on only one side in each subject which corresponded with gene expression Subject increased α -SG by fold on the side of gene transfer reaching wild-type levels This was accompanied by an increase in mean muscle ber diameter, 28.2 ± 11.1 to 52.2 ± 13.1 Subject demonstrated robust gene expression, again equivalent to wild-type levels on the side of gene transfer MHCI and CD4, CD8 mononuclear cells were upregulated on the side of gene transfer in subjects and ELISpots for antigen specic production of IFN-γ secretion were monitored beginning pre-gene transfer Subject had a transient minimal response to AAV1 capsid pools at days 14 and 28 Patient showed no response to α-SG or AAV1 capsid peptide pools In contrast to the rst two patients, Subject had pre-existing immunity to AAV1 (neutralizing antibodies) and an ELISpot IFN-γ response within the rst week to AAV following gene transfer suggesting a memory response Levels of gene expression were also reduced in this subject compared to others Overall the promising results from this study with gene expression persisting for months lays the foundation for potentially achieving long-term sustained correction of the underlying gene defect in muscular dystrophy However, the inuence of pre-existing immunity to AAV may affect initial muscle ber transduction or sustained gene expression 291 Advanced Generation RGD-Modied Oncolytic Adenovirus ICOVIR-7 in the Treatment of Cancer Refractory to Other Modalities Petri Nokisalmi,1,2 Sari Pesonen,1,2 Sophie Escutenaire,1,2 Mari Raki,1,2 Vincenzo Cerullo,1,2 Ramon Alemany,3 Juanjo Rojas,3 Maria Rajecki,1,2 Lotta Kangasniemi,4 Anna Kanerva,1,5 Timo Joensuu,6 Laura Ahtiainen,1,2 Akseli Hemminki.1,2 Cancer Gene Therapy Group, Transplantation laboratory & Haartman Institute & Finnish Institute for Molecular Medicine, University of Helsinki, Helsinki, Finland; 2HUSLAB, Helsinki University Central Hospital, Helsinki, Finland; 3Translational Research Laboratory, IDIBELL-Institut Català d’Oncologia, L’Hospitalet de Llobregat, Barcelona, Spain; 4Oncos Therapeutics Inc., Helsinki, Finland; 5Department of Obstetrics and Gynecology, Helsinki University Central Hospital, Helsinki, Finland; 6International Comprehensive Cancer Center Docrates, Helsinki, Finland and 7/15 of samples were positive several weeks later suggesting virus replication Overall, objective evidence of anti-tumor activity was seen in 9/17 evaluable patients Clinical benet in radiological analysis was seen in 5/12 evaluable patients, consisting of one partial response, two minor responses and two cases of stable disease In addition, 3/3 evaluable patients had a decrease in tumor density In summary, ICOVIR-7 treatment seems safe, results in anti-cancer activity and is therefore promising for further clinical testing 292 Final Results of Phase I Trial of GMCSF Gene-TGFȕ2 Antisense Gene Autologous Tumor Cell (TAG) Vaccine in Advanced Cancer John Nemunaitis,1,2,3,4,5 Padmasini Kumar,5 Neil Senzer,1,2,3,4,5 Joseph Kuhn,6 Minal Barve,1 Jairo Olivares,1 Gerald Edelman,1 Cynthia Bedell,1 Yang Yu,5 Mitchell Magee,7 Beena O Pappen,5 Gladice Wallraven,5 Alex W Tong,5 Phillip B Maples.5 Mary Crowley Cancer Research Centers, Dallas, TX; 2Medical City HCA, Dallas, TX; 3Baylor Sammons Cancer Center, Dallas, TX; 4Texas Oncology, P.A., Dallas, TX; 5Gradalis, Inc., Dallas, TX; General and Oncology Surgery Associates, Dallas, TX; 7Cardio Thoracic Surgery Associates of North Texas, Dallas, TX Both GMCSF gene transduced vaccine and TGFβ2 antisense gene vaccine, in separate trials, have demonstrated benecial effects without evidence of toxicity in advanced cancer patients The GMCSF transgene directly stimulates increased expression of tumor antigen(s) and enhances dendritic cell migration to the vaccination site TGFβ2 blockade following intracellular TGFβ2 antisense gene expression reduces production of immune inhibiting activity at the vaccine site We recently completed a phase I clinical investigation of the combined GMCSF/TGFβ2 antisense TAG vaccine under BB IND 13650 Nineteen patients [6/13, M/F; median age 58 yrs; median number prior chemotherapy regimens 2; cancer: melanoma (3), lung (2), colon (4), breast (2), neuroendocrine (4), other (4)] have received between and 12 vaccinations Harvested malignant tissue was processed to a single cell suspension, and cells were transfected via electroporation with our novel TAG expression vector and irradiated (median number vaccines/patient, 10) Patients received monthly intradermal injections in two dosing cohorts (1x107 cells/ injection or 2.5x107 cells/injection) for a minimum of months No grade 3, toxic events related to therapy were observed in the 19 treated patients One patient was unevaluable for response due to discontinuation after one month related to travel difculty Sixteen of 18 patients maintained stable disease for months Two patients maintained stable disease for one year; a third (melanoma) achieved complete response (CR) and remains in CR now 18 months after rst vaccination (Figure 1).Figure We treated 21 cancer patients with a single round of oncolytic adenovirus ICOVIR-7 with doses ranged from x 1010 VP to x 1012 VP All patients had advanced and metastatic solid tumors refractory to standard therapies ICOVIR-7 features an RGD-4C modication of the ber HI-loop of serotype adenovirus for enhanced entry into tumor cells Tumor selectivity is mediated by an insulator, a modied E2F promoter and an Rb binding site deletion, while replication is optimized with E2F binding hairpins and a Kozak sequence ICOVIR-7 treatment was well tolerated with mild to moderate fever, fatigue, elevated liver transaminases, chills and hyponatremia One patient had grade anemia but no other serious side effects were seen At baseline, 8/20 of evaluable patients had neutralizing antibody titers (NAb) against the ICOVIR-7 capsid Treatment resulted in NAb induction within four weeks in 16/17 patients No elevations of serum pro-inammatory cytokines were detected Viral genomes were detected in the circulation in 18/21 of patients after injection Molecular Therapy Volume 18, Supplement 1, May 2010 Copyright © The American Society of Gene & Cell Therapy S111 CLINICAL GENE & CELL THERAPY ORAL ABSTRACT SESSION DNR-transduced CTL were resistant to the anti-proliferative effects of recombinant TGF-ß in vitro Both patients received x 107CTLs/m2 without toxicity Neither patient received additional therapy following these CTL infusions DNR-transduced T-cells have been detected by quantitative real-time PCR amplication in the peripheral blood of both patients for >3 and >6 months respectively post infusion One patient had a complete remission and the other has a mixed clinical response Hence, immunotherapy with CTL targeting LMP antigens is well tolerated in patients with EBV+ lymphoma and can induce durable clinical responses Furthermore, for patients resistant to CTL therapy, the use of DNR-transduced CTL may be benecial 294 Efcient Engraftment of MGMTP140K Gene-Modied CD34+ Cells Following Nonmyeloablative BCNU Conditioning in Patients with Glioblastoma Conclusion: GMP manufacturing of TAG vaccine is feasible, safety is achieved and evidence of clinical benet is demonstrated 293 Administration of Tumor-Specic Cytotoxic T Lymphocytes Engineered to Resist TGF-ß to Subjects with EBV-Associated Lymphomas Catherine M Bollard,1 Gianpietro Dotti,1 Stephen Gottschalk,1 Enli Liu,1 Andrea M Sheehan,2 George Carrum,1 Hao Liu,1 Chung-Che Chang,3 Adrian P Gee,1 Malcolm K Brenner,1 Helen E Heslop,1 Cliona M Rooney.1 Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, TX; 2Department of Pathology, Texas Children’s Hospital, Houston, TX; 3The Methodist Hospital and TMHRI, Weill Medical College of Cornell University, Houston, TX EBV-associated Hodgkin’s Lymphoma (HL) and some nonHodgkin’s lymphomas (NHL) express EBV proteins which are potential targets for immunotherapy Lymphomas arising in the immune-competent host, however, utilize multiple immune evasion strategies to evade host T cell responses These include limited expression of EBV antigens, so that only the subimmunodominant EBNA1 and LMP1 and antigens are expressed (type II latency); and production of immunosuppressive cytokines, such as TGF-ß We rst tested our hypothesis that cytotoxic T lymphocytes (CTL) enriched for specicity to LMP antigens would have efcacy in patients with EBV+ lymphoma LMP-CTL were generated from 29 patients with EBV+ HL and NHL using antigen-presenting cells genetically modied to overexpress either LMP2 alone (n=16), or LMP2 and inactive LMP1 (∆LMP1) (n=14) No immediate toxicity was observed following infusion 14/15 high-risk and/or multiply relapsed patients who received LMP-CTL as adjuvant treatment remain in remission for a median of 2.5 years (range months to >5 years) Of the 14 patients who had detectable disease at the time of CTL infusion, 11 had clinical responses (78%), including (57%) complete responses The median duration of the clinical responses is 1.5 years However, 36% of patients were either resistant to CTL therapy (n=3) or relapsed/ progressed >9 months after CTL therapy (n=2), suggesting the importance of additional tumor immune escape mechanisms TGF-ß inhibits the growth and function of effector T cells and is secreted both by lymphoma cells and by inltrating T regulatory cells To test our hypothesis that the infused CTL can be made resistant to the TGF-ß produced by these tumors, LMP-CTL from patients with relapsed EBV+ve HL one of whom had failed initial CTL therapy, were transduced with a retrovirus vector encoding the dominantnegative TGF-ß type II receptor (DNR) Unlike non-transduced CTL, S112 Jennifer E Adair,1 Brian C Beard,1 Alexander Spence,2 Jason Rockhill,2 Daniel L Silbergeld,2 Robert Rostomily,2 Pamela Becker,2 Marc Chamberlain,2 Maciej Mrugala,2 Hans-Peter Kiem.1,2 Fred Hutchinson Cancer Research Center, Seattle, WA; University of Washington, Seattle, WA Glioblastoma, the most common primary malignant brain tumor in adults, confers poor prognosis, with a median survival of 15 months, notwithstanding aggressive treatment Furthermore, patients whose tumors present with an unmethylated methylguanine methyltransferase (MGMT) promoter display a poorer prognosis than patients with methylated MGMT promoter In an effort to improve therapy and outcome for patients with unmethylated MGMT, and thus high wild type MGMT expression, chemotherapy combining alkylating agents carmustine (BCNU) or temozolomide (TMZ) with the wild type MGMT active site inhibitor O6-benzylguanine (O6BG) has been used, but has been associated with dose limiting hematopoietic toxicity We have previously demonstrated in the dog and nonhuman primate model efcient gene marking, in vivo selection and chemoprotection of hematopoietic cells from O6BG and BCNU or TMZ following transplantation with MGMTP140K gene-modied CD34+ cells Based on these ndings, we have developed a clinical gammaretroviral vector produced by Phoenix-GALV packaging cells for use in a Phase I/II clinical trial This study (National Clinical Trials Registry: NCT00669669) enrolled three glioblastoma patients following surgical resection, diagnosis and pathological subtyping of unmethylated tumor MGMT promoter status in each patient Enrolled patients underwent standard radiation therapy, followed by G-CSF mobilization, apheresis and conditioning with 600 mg/ m2 BCNU prior to infusion of gene-modied cells Gene marking in pre-infusion colony forming units (CFUs) was 70.6%, 79.0% and 74.0% in patients 1, and respectively, as determined by CFU-PCR The dose of BCNU resulted in patient neutrophil counts below 500/ µL for only 2, and days, respectively, in the three patients, and no platelet count below 28,000/µL Following engraftment, initial gene marking in peripheral blood mononuclear cells (PBMNCs) and sorted granulocytes ranged between 0.37 to 0.84 and 0.33 to 0.83 provirus copies, respectively, by real-time PCR Intracellular MGMT-staining of PBMNCs confirmed MGMT expression Furthermore, gene marking in CFUs from CD34-selected cells at 1-2 months post-transplant ranged from 28.5% to 47.4% Initial integration site analysis in Patients and has demonstrated that multiple gene-modied clones are contributing to hematopoiesis and is in process for Patient All three patients have been treated with at least one cycle of O6BG and TMZ post-transplant with preliminary evidence of in vivo selection of gene-modied cells No additional extra-hematopoietic toxicity has been observed in any patient to date We believe that these data demonstrate the feasibility of achieving signicant engraftment of MGMTP140K- modied cells with a wellMolecular Therapy Volume 18, Supplement 1, May 2010 Copyright © The American Society of Gene & Cell Therapy ... mobilization, apheresis and conditioning with 600 mg/ m2 BCNU prior to infusion of gene- modied cells Gene marking in pre-infusion colony forming units (CFUs) was 70.6%, 79.0% and 74.0% in patients... clinical gammaretroviral vector produced by Phoenix-GALV packaging cells for use in a Phase I/ II clinical trial This study (National Clinical Trials Registry: NCT00669669) enrolled three glioblastoma... Engraftment of MGMTP140K Gene- Modied CD34+ Cells Following Nonmyeloablative BCNU Conditioning in Patients with Glioblastoma Conclusion: GMP manufacturing of TAG vaccine is feasible, safety is achieved