ImmunoTACE TRIAL A Randomised phase II Clinical Trial of conditioning cyclophosphamide and Chemoembolisation with or without Vaccination with Dendritic Cells pulsed with HepG2 lysate ex vivo in Patients with Hepatocellular Carcinoma EUDRACT NUMBER: 2011-001690-62 PROTOCOL VERSION: 9.0a 15-Jul-2019 PROTOCOL NUMBER: HE2016 ISRCTN: 11889464 SPONSOR NUMBER: RG_10-148 CHIEF INVESTIGATOR: Professor David Adams University of Birmingham 1 of 65 ImmunoTACE_Protocol_v9.0a_15-Jul-2019 CHIEF INVESTIGATOR SIGNATURE PAGE ImmunoTACE TRIAL A Randomised phase II Clinical Trial of conditioning cyclophosphamide and Chemoembolisation with or without Vaccination with Dendritic Cells pulsed with HepG2 lysate ex vivo in Patients with Hepatocellular Carcinoma Version 9.0a 15-Jul-2019 This Protocol has been approved by: Professor David Adams Signature : Date : Chief Investigator This protocol describes the ImmunoTACE trial and provides information about procedures for patients taking part in the ImmunoTACE trial. The protocol should not be used as a guide for treatment of patients not taking part in the ImmunoTACE trial. This protocol was written using CRCTU-PRT-QCD-001, version 1.0 2 of 65 ImmunoTACE_Protocol_v9.0a_15-Jul-2019 SPONSOR University of Birmingham TRIAL PERSONNEL CHIEF INVESTIGATOR Professor David Adams*# Consultant Hepatologist and University of Dean of Medicine Birmingham1 CLINICAL INVESTIGATOR Dr Yuk Ting Ma# Senior Clinical Lecturer in University of Hepatobiliary Oncology Birmingham2 CO-INVESTIGATORS Dr Stuart M Curbishley* Senior Research Fellow University of Birmingham2 LEAD STATISTICIAN Dr Christina Yap Professor of Clinical Trial The Insitute of Cancer Biostatistics Research3 TRIAL STATISTICIAN Miss Amanda Kirkham* Biostatistician University of Birmingham2 TRIAL MANAGEMENT Dr Anna Rowe* Trial Management Team Leader University of TEAM Birmingham2 Dr Jessica Douglas* Senior Trial Coordinator University of Birmingham2 Ms Camilla Bathurst Trial Coordinator University of Birmingham2 # Designated clinical coordinators for the clinical trial 1 NIHR Birmingham Liver Biomedical Research Centre, University of Birmingham, Birmingham. B15 2TT 2 Cancer Research UK Clinical Trials Unit, Institute of Cancer and Genomic Sciences, University of Birmingham, Birmingham. B15 2TT 3 The Institute of Cancer Research, 15 Cotswold Road, Sutton, SM2 5NG Contact Details: Email: [email protected] Telephone: 0121 371 8117 or 0121 371 8158 3 of 65 ImmunoTACE_Protocol_v9.0a_15-Jul-2019 AMENDMENTS The following amendments and/or administrative changes have been made to this protocol since the implementation of the first approved version Protocol Type of Amendment Amendment Date of Version (e.g. substantial/ Summary of amendment? No. Amendment No. Non-substantial/ administrative change) 1 21_Nov_2013 n/a Substantial - Change of PI at site - Change to study personnel, contact details and 2 02_Mar_2015 5.0 Substantial change of CI - Typographical errors corrected - Updated Abbreviated List 3 05_Oct_2015 6.0 Substantial - Change to study personnel, and contact details - Typographical errors corrected - RECIST criteria updated to v1.1 throughout - Section 3 – Treatment Details (Autologous Dendritic Cells Pulsed with HepG2 Lysate matured with Monophosphoryl Lipid A) N/A 22_Mar_2016 7.0 N/A Protocol was updated and signed off internally, but not submitted to the Regulatory Authorities. This was subsequently further amended and any changes included in V8.0 (see details in row below) 4 17_Nov_2016 8.0 Substantial - Schedule updated to match text (and vice versa) and footers clarified - Day of Leukapheresis moved to prior to Day 1 Cyclophosphamide - Randomisation timelines clarified - Addition of 3 month survival status follow-up - 3.1 Group 2 patients updated information - 4.2 clarified windows for visits and progressive disease in first 12 months - 6.2 Screening – HLA typing removed - 6.3 Contraception section added - 7.2 Randomisation – text updated - 8.0 Treatment and follow-up – updated to 4 of 65 ImmunoTACE_Protocol_v9.0a_15-Jul-2019 match schedule - 9.0 Dose Modifications – clarified - 10 Leukapheresis procedure time updated - 11 Evaluable patient term clarified - 12.1.4 text updated - 12.2 Reporting periods clarified - 12.3 Reporting procedures clarified and SAE contact details updated - 13 Updated and clarified - 16.3 Sample size calculation updated - 16.4 Analysis plan updated - Amendment table streamlined - Change to trial personnel and contact details - Terminology and typographical errors clarified or corrected 5 16_Aug_2018 9.0 Substantial - Change to study personnel and contact details - 16.3 Change in power calculation to allow for a reduction in sample size from 70 to 48 with a slight reduction in power, taking into account a longer recruitment period - 16.1, 16.2 clarified Primary outcome and provide further details on primary analysis - Trial synopsis, 16.1 and 16.2 added mRECIST criteria as a secondary outcome measure - Trial synoposis & 5.1 inclusion criteria updated - 16.4 amended main analysis set to modified intention-to-treat - 18 & 19 Data Protection sections amended to incorporate the General Data Protection Regulation 2018 and Data Protection Act 2018 - Screening window increased and screening period clarified - Window added for D31 TACE - Window added for D62, 92 and 122 DC Vaccine - Follow-up visit window re-defined 6 15-Jul-2019 9.0a Non-Substantial - Change to trial personnel & contact details - Administrative errors corrected for: 1. Serology Tests: Trial Schedule, 6.2 & 8.1 5 of 65 ImmunoTACE_Protocol_v9.0a_15-Jul-2019 2. End of Trial Definition: Trial Schedule, 8.1 & 15 3. eGFR: Trial Schedule, 6.2 & 8.1 6 of 65 ImmunoTACE_Protocol_v9.0a_15-Jul-2019 TRIAL SYNOPSIS Title A randomised phase II clinical trial of conditioning cyclophosphamide and chemoembolisation with or without vaccination with dendritic cells (DC) pulsed with HepG2 cell lysate ex vivo in patients with hepatocellular carcinoma (HCC). Background A previous study by the Liver Research Team (Birmingham) using DC pulsed ex vivo with the lysate of the HepG2 cell line has shown clinical benefit with evidence of antigen-specific T-cell responses in some patients with advanced HCC. This project intends to investigate the activity of this vaccine in combination with chemoembolisation compared to chemoembolisation alone in patients with intermediate stage HCC. All patients will receive a conditioning regimen comprising low dose cyclophosphamide. Study Design Randomised, open label, multicentre, phase II trial Objectives To determine whether activity due to the addition of DC vaccine to chemoembolisation and preconditioning cyclophosphamide prolongs progression free survival (PFS) and warrants further investigation Recruitment/Patient Population Patients over the age of 18 years, with HCC with performance status 0 - 2, with adequate renal function and hepatic function and in Child-Pugh category A or B and considered suitable for chemoembolisation. There should be no active concurrent infections or other malignancies and the patients must not be taking immunosuppressive therapy. Patients must not be immunocompromised. Planned Interventions Group 1: TACE therapy + preconditioning cyclophosphamide only Group 2: TACE therapy + preconditioning cyclophosphamide + dendritic cells infusions. Outcome Measures Primary outcome measure: 1. Progression free survival time Secondary outcome measures: 1. Radiological response (RECIST 1.1 criteria) 2. Change in the tumour marker serum AFP 3. Assessment of toxicity using NCI-CTCAE version 4 4. Immune response 5. Overall survival time 6. Radiological response based on modified RECIST (mRECIST) 7. Progression free survival at 12 months where progression is determined by mRECIST Proposed Sample Size A total of 48 (24 in each group) patients with HCC will be recruited. 7 of 65 ImmunoTACE_Protocol_v9.0a_15-Jul-2019 TRIAL SCHEDULE Screening Leukapheresis Day 1 Day 29 Day 31 Day 38, 45 & Day 60, 90 & Day 62, 92 & Day 152, 242, Survival Status 52 120 122 332 & 422 Follow-up ¥ Cy infusion Cy TACE visit Post-TACE Repeat Cy Repeat DC Follow-up visits Three monthly infusion follow-up visits infusion visits infusion visits ALL Group 2 ALL ALL ALL ALL ALL Group 2 ALL ALL patients ONLY patients patients patients patients patients ONLY patients patients Between 2 + 3 days after 14 + 7 days from Up to 14 after 2 + 3 days Visit Window (days) Randomisation +/- 3 +/- 3 +/- 3 Day 60, 90 & +/- 10 +/- 10 Randomisation Randomisation after Day 29 and Day 1* 120 Informed Consent X Medical History X Physical Examination X1 X X X X ECOG Performance Status X X X X X X X X X Concomitant Medications X X X X X X X X X Adverse Events X X X X X X X X X Clinical Events X X X X X X X X X Vital Signs X X X X X X X X X Mandatory Microbiology screening2 X X X Tumour Assessment3 X X (DAY 60 ONLY) Auto-Antibody Tests4 X X X X Standard Blood Tests5 X X X X X X X Immune Response Blood Tests X X X X X Blood Collection for Generation of X DC Vaccine6 via Leukapheresis Pregnancy Test7 X 8 of 65 ImmunoTACE_Protocol_v9.0a_15-Jul-2019 Electrocardiogram (ECG) X Height and Weight X Dendritic Cell Infusion8 X X Cyclophosphamide Infusion X X X Survival Status9 X 1. This will include assessment for potential difficulties in intra-hepatic artery cannulation (i.e. severe atherosclerotic disease) and venous access for leukapheresis at screening. 2. All patients will require mandatory testing for blood borne infectious agents of blood products prior to processing and storage. These tests