Adoptive Transfer of iNKT Cells for Treating Patients With Relapsed/Advanced HCC

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.

• iNKT cells (Biological): 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.
• IL-2 (Drug), also known as Human recombinated Interleukin-2: IL-2 will be given at a dose of 25,000 IU/kg/day for 5-14 days.
• Tegafur (Drug), also known as Gimeracil and Oteracil Potassium Capsules: Tegafur will be given at a dose of 40\~60 mg bis in die (BID) 2 weeks.

Primary outcomes

• Number of Adverse Events [During the first 12 weeks, participants were assessed for adverse events every 2-4 weeks after infusion; after the first 12 weeks, participants were assessed for adverse events every 3 months, up to 20 months.]: Defined as signs/symptoms, laboratory toxicities, and clinical events that are possibly, likely, or definitely related to study treatment Adverse events assessed according to NCI-CTCAE v4.0 criteria 2.

Secondary outcomes

• Progression-Free Survival (PFS) [Through study completion, an average of 12 months.]: PFS is the time that passes from the date that patient enrolled in the clinical trial and the date on which HCC progresses or the date on which the patient dies. HCC progression was evaluated by imaging according to the irRC standard. Progression is defined as a ≥25% increase in the nadir of the sum of target lesions.
• Number of Participants With Stabilized (SD) or Progressive (PD) Disease. [4 weeks, 8 weeks, 12 weeks and 24 weeks after cell infusion.]: Disease stabilization (SD) or progressive diseasee (PD) were valuated by imaging according to the irRC standard. Complete response (CR): Disappearance of all lesions; Partial response (PR): ≥50% decrease from baseline; SD: Neither CR or PD is met; PD≥25% increase in the nadir of the sum of target lesions.
• Overall Survival (OS) [Through study completion, up to 20 months.]: OS is the time that passes from the date that patient enrolled in the clinical trial and the date on which the patient dies, according to the irRC standard.

Sex: All, ages: 18 Years to 80 Years

Inclusion Criteria:

* Age 18-80 years.
* Patients with hepatocellular carcinoma (BCLC, stage C) proved by histopathology or proved by CT or MRI imaging system, relapsed after previous therapy and no effective therapies known at this time.
* Life expectancy of ≥ 12 weeks.
* WBC>3.5×10\^9/L, LYMPH> 0.8×10\^9/L, Hb>85g/L, PLT>50×10\^9/L, Cre<1.5×the upper limit of normal value.
* iNKT>10/mL in peripheral blood mononuclear cell (PBMC).
* Able to understand and sign the informed consent.

Exclusion Criteria:

* Any uncontrolled systematic disease: hypertension, heart disease, and et al.;
* Portal vein tumor thrombus, central nervous system tumor metastasis, or combined with other tumors;
* Receiving radiochemotherapy, local therapy, or targeting drugs within 4 weeks prior to this treatment;
* Unstable immune systematic diseases or infectious diseases;
* Combined with AIDS or syphilis;
* Patients with history of stem cell or organ transplantation;
* Patients with allergic history to related drugs and immunotherapy;
* Patients with complications associated with liver diseases: moderate or severe pleural effusion, pericardial effusion, ascites, or gastrointestinal hemorrhage;
* Pregnant or lactating subjects;
* Unsuitable subjects considered by clinicians.

Participant flow

Outcome results

• Number of Adverse Events (Primary), Number, events: iNKT Cell Loading Dose:3x10^7/m2 12; iNKT Cell Loading Dose:6x10^7/m2 2; iNKT Cell Loading Dose:9x10^7/m2 8; iNKT Cell Loading Dose:1x10^10/m2 4
• Progression-Free Survival (PFS) (Secondary), Count of participants, Participants: iNKT Cell Loading Dose:3x10^7/m2 1; iNKT Cell Loading Dose:6x10^7/m2 1; iNKT Cell Loading Dose:9x10^7/m2 2; iNKT Cell Loading Dose:1x10^10/m2 0
• Number of Participants With Stabilized (SD) or Progressive (PD) Disease. (Secondary), Count of participants, Participants: iNKT Cell Loading Dose:3x10^7/m2 3; iNKT Cell Loading Dose:6x10^7/m2 3; iNKT Cell Loading Dose:9x10^7/m2 3; iNKT Cell Loading Dose:1x10^10/m2 1
• Overall Survival (OS) (Secondary), Number, participants: iNKT Cell Loading Dose:3x10^7/m2 1; iNKT Cell Loading Dose:6x10^7/m2 0; iNKT Cell Loading Dose:9x10^7/m2 2; iNKT Cell Loading Dose:1x10^10/m2. 0

Adverse events

Time frame: During the first 12 weeks, participants were assessed for adverse events every 2-4 weeks after infusion; after the first 12 weeks, participants were assessed for adverse events every 3 months, up to 20 months.

Serious adverse events by term

Other adverse events by term

• Study Protocol and Statistical Analysis Plan (PDF, 2017-03-01)

• Jun Lu, Director, Study chair, Beijing YouAn Hospital