Clinical trial NCT03175679
Conditions: Hepatocellular Carcinoma
Sponsor: Beijing YouAn Hospital
Start date: 2017-04-01; Completion date: 2019-03-30
Enrollment: 10 participants; Allocation: NON_RANDOMIZED; Intervention model: SINGLE_GROUP

Brief summary:
This study enrolls patients who have relapsed/advanced hepatocellular carcinoma (HCC, BCLC stage C). The HCC tumor relapsed or metastasized through the body after standard treatment or the patients cannot receive standard treatment under current conditions. This research study uses special immune system cells called iNKT cells, a new experimental treatment.

The purpose of this study is to find the biggest dose of iNKT cells that is safe and tolerance, to see how long they last in the body, to learn the immunoresponse in the body, to learn the side effects are and to see if the iNKT cells will help people with relapsed/advanced hepatocellular carcinoma (HCC).

Detailed description:
PBMCs of enrolled patients were collected and then further separated by density gradient centrifugation. After washing three times, the cells were resuspended in serum-free medium with recombinant human IL-2 and α-GalCer. Restimulation with α-GalCer-pulsed autologous DCs was done on days 7. After 14 days of cultivation, the iNKT cells were harvested, washed thrice, and then resuspended in saline. The frequency of iNKT cells before and after cultured in vitro were determined by flow cytometry analysis.

Arms: arm 1: Autologous in vitro expanded iNKT cells in conjunction with IL-2 and along with lymphodepleting chemotherapy (Tegafur) will be administered to patients with advanced HCC.

iNKT Cell Loading Dose：3x10\^7/m2.

IL-2: IL-2 will be given at a dose of 25,000 IU/kg/day for 5-14 days.

Tegafur: Tegafur will be given at a dose of 40\~60 mg bis in die (BID) 2 weeks. arm 2: Autologous in vitro expanded iNKT cells in conjunction with IL-2 and along with lymphodepleting chemotherapy (Tegafur) will be administered to patients with advanced HCC.

iNKT Cell Loading Dose：6x10\^7/m2.

IL-2: IL-2 will be given at a dose of 25,000 IU/kg/day for 5-14 days.

Tegafur: Tegafur will be given at a dose of 40\~60 mg bis in die (BID) 2 weeks. arm 3: Autologous in vitro expanded iNKT cells in conjunction with IL-2 and along with lymphodepleting chemotherapy (Tegafur) will be administered to patients with advanced HCC.

iNKT Cell Loading Dose：9x10\^7/m2.

IL-2: IL-2 will be given at a dose of 25,000 IU/kg/day for 5-14 days.

Tegafur: Tegafur will be given at a dose of 40\~60 mg bis in die (BID) 2 weeks. arm 4: Autologous in vitro expanded iNKT cells in conjunction with IL-2 and along with lymphodepleting chemotherapy (Tegafur) will be administered to patients with advanced HCC.

iNKT Cell Loading Dose：1x10\^10/m2.

IL-2: IL-2 will be given at a dose of 25,000 IU/kg/day for 5-14 days.

Tegafur: Tegafur will be given at a dose of 40\~60 mg bis in die (BID) 2 weeks.
Interventions: intervention 1: iNKT cells intervention 2: IL-2 intervention 3: Tegafur
Intervention descriptions: intervention 1: The patients received different doses of in vitro-expanded autologous iNKT cells through intravenous infusion. The dosage was escalated from 3x10\^7 cells/m2 to 6x10\^7 cells/m2 to 9x10\^7 cells/m2. The maximum tolerated dose (MTD) was defined as the last dose level, and the dosage would be up to 1x10\^10 cells/m2 if no MTD was observed after a 3+3 design. intervention 2: IL-2 will be given at a dose of 25,000 IU/kg/day for 5-14 days. intervention 3: Tegafur will be given at a dose of 40\~60 mg bis in die (BID) 2 weeks.

Protocol documents:
Prot_SAP_000.pdf: 
A Study of Adoptive Invariant Nature Killer T Cell Therapy for 
Relapsed/Advanced Hepatocellular Carcinoma (HCC)   
 
NCT03175679  
March 1, 2017 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 


Background 
Invariant Natural Killer T (iNKT) cells activated by a specific glycolipid 
antigen (a-galactosylceramide) show strong antitumor activity against 
malignant tumor through producing high levels of cytokines, such as IFN-
γ and IL-4. iNKT cells and their frequencies are abundant in the liver, but 
defect in malignant tumor development. We did a phase I dose escalation 
study with autologous in vitro expanded iNKT cells in advanced 
hepatocellular carcinoma (HCC). 
Objectives 
Inclusion criteria included diagnosed of HCC by CT or MRI, BCLC B/C 
stage, age 18 to 80 and predicted life span of 12 weeks or more to complete 
the study and follow-up. Exclusion criteria included uncontrolled systemic 
diseases, central nervous system metastases or other tumor, unstable 
immune system disease, positive HIV, syphilis or immune deficiency 
disease and known hypersensitivity immunotherapy and related drugs. All 
patient understood and voluntarily signed the informed consent. 
Method 
Patients underwent leukapheresis to collect circulating iNKT cells, which 
were isolated using a monoclonal antibody (6B11) against the invariant 
TCR-α chain expressed by iNKT cells and expanded in culture over 14 
days. The expanded cells were harvested and divided into 3 doses. The 
expansion rates of iNKT cells in enrolled patients before and after 


cultivation were analysed by flow cytometry analysis. Realtime-PCR was 
conducted to compare the mRNA level of Th1 and Th2 cytokines before 
and after iNKT cultivation. 
Stuty design 
Patients were intravenously doped with different doses of autologous iNKT 
cells, 3 infusions per course. Dosages were escalated from 3×107 cells/m2 
to at least 9×107cells/m2. Tegafur will be given twice per day for 2 weeks 
after apheresis. Human IL-2 will be given during 5-14 days after iNKT 
cells infusion. Subjects were monitored for 4 weeks post-infusion to make 
sure the safety of this therapy. The laboratory safety tests and physical 
examination were carried out at 2nd week, 4th week, 8th week and 12th 
week after iNKT infusion. All subjects were followed every 3 months until 
the disease progressed. 
 
Statistical Analysis Plan 
Means and standard deviations was used to analyze difference of expansion 
rates of iNKT cells and mRNA level of Th1 and Th2 cytokines in enrolled 
patients before and after cultivation, differences were considered 
significant for p < 0.05.