Safety Study of Liver Natural Killer Cell Therapy for Hepatoma Liver Transplantation

Study Overview
=================
Brief Summary
-----------------
The objective of this study is to evaluate feasibility and safety of the adoptive transfer of activated natural killer (NK) cells extracted from cadaveric donor liver graft perfusate for liver transplant recipients with hepatocellular carcinoma (HCC)

Detailed Description
-----------------
To investigate whether adoptive transfer of IL-2 stimulated NKcells extracted from cadaveric donor liver graft exudates into recipient with HCC will lead to prevent recurrence of HCC after liver transplantation. The method we plan to use is to collect NK cells from the cadaveric donor liver perfusate at the time of organ recovery. The perfusate will be condensed by centrifuging and liver mononuclear cells (LMNC) were isolated by gradient centrifugation. LMNC will be cultured with anti-CD3 antibody. LMNC will be separted into a CD3-/CD56+ NK cell fraction and a non-NK cell fraction. The purity of isolated fractions will be assessed by Flow cytometric analyses. NK cells will be cultured with human recombinant IL-2 in 10% RPMI at 37 ºC in a 5% CO2 incubator. NK cells will be infused intravenously to the recipient with HCC who have liver transplant from the same cadaveric donor at post operative day 4 in attempt to prevent the metastasis and recurrence of HCC.

Official Title
-----------------
A Novel Immunotherapy for Liver Transplant Patients With Hepatocellular Carcinoma: Anti-tumor Effect of IL2-activated Donor Liver Natural Killer Cell

Conditions
-----------------
Liver Cirrhosis, Hepatocellular Carcinoma, Evidence of Liver Transplantation

Intervention / Treatment
-----------------
* Biological: Liver NK cell inoculation


Participation Criteria
=================
Eligibility Criteria
-----------------
Inclusion Criteria: Primary liver transplant recipient with HCC Patients need to meet the liver transplant eligibility criteria Cardiac function; cardiac echo will indicate that ejection fraction (EF) > 35% or right ventricular systolic pressure (RVSP) < 50mmHg. Stress echo will show no ischemic lesion (if applicable). Pulmonary function; SpO2 >90% or PaO2 > 60 mmHg, or CT will show no active pulmonary lesion. Complete blood count; Platelet > 20,000 /mm^3, Hematocrit > 20%, WBC > 1,000 /mm^3 Eighteen years of age or older Ability to provide informed consent If female of childbearing potential: Must not be lactating, must have a negative serum B-human chorionic gonadotropin (HCG) test within 7 days prior to Day of Transplant, and must agree to practice an acceptable and reliable form of contraception during the study Ability to provide informed consent Exclusion Criteria: Living donor liver transplant; a healthy person donates part of his or her liver to the recipient Multiple organ transplants Prior solid organ or bone marrow transplant recipients Fluminant hepatic failure The patients regularly receive the hemodialysis more than twice a week before liver transplant Status 1 transplants; acute severe disease and defined as a patient with only recent development of liver disease who is in the intensive care unit of the hospital with a life expectancy without a liver transplant of fewer than 7 days ABO incompatible transplants Transplants utilizing livers from non-heart beating donors; the cardiac death donor Recipients of investigational therapy within 90 days prior to transplant procedure Acute viral illness History of malignancy within 5 years, with exception of: Adequately treated localized squamous or basal cell carcinoma of the skin without evidence or recurrence, and/or Hepatocellular carcinoma Illness other than primary liver disease (e.g., severe ischemic heart disease, left ventricular dysfunction, or pulmonary disease), which, in the opinion of the Investigator, may significantly increase the risk of the transplantation procedure Current drug or alcohol abuse or, in the opinion of the Investigator, is at risk for poor compliance (no drug testing required) Serology positive donor (HCV, HBsAg, HBcAb, HTLV-1, HTLV-3, EBVIgM) Poor liver function donor (Total bilirubin > 3.0 mg/dl, Prothrombin time > 35 sec), Patients who receive intercurrent chemotherapy at the time of enrollment

Ages Eligible for Study
-----------------
Minimum Age: 18 Years

Sexes Eligible for Study
-----------------
All

Accepts Healthy Volunteers
-----------------
No

Study Plan
=================
How is the study designed?
-----------------

Design Details

Primary Purpose: Treatment
Allocation: Non-Randomized
Intervention Model: Parallel Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Small dose<br>From the donor liver perfusate, mononuclear cell will be extracted and cultured. Then, the cells will be stimulated with IL-2. The number of inoculation cells( mainly NK cells) is between 10 and 100 million cells. The cells will be given to the liver transplant recipient who had the same donor for liver and liver perfusate. Patient of this arm receive small dose of liver NK cell inoculation as described. | Biological: Liver NK cell inoculation<br>* Liver transplant recipients will receive once liver NK cell inoculation several days after liver transplantation.<br>|
| Experimental: Large dose<br>From the donor liver perfusate, mononuclear cell will be extracted and cultured. Then, the cells will be stimulated with IL-2. The number of inoculation cells(mainly NK cells) is between 100 and 1000 million cells. The cells will be given to the liver transplant recipient who had the same donor for liver and liver perfusate.Patient of this arm receive large dose of liver NK cell inoculation as described. | Biological: Liver NK cell inoculation<br>* Liver transplant recipients will receive once liver NK cell inoculation several days after liver transplantation.<br>|

What is the study measuring?
-----------------
Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Side Effect of Cadaveric Donor Liver NK Cell Infusion | Side effect of cadaveric donor liver NK cell infusion We will measure the occurence of the side effect of the liver NK cell infusion. We will monitor the patient condition clinically. If any side effect are noticed, we will record them and report to the data safety monitoring comittee. | 1 year | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| NK Cell Infusion-related Toxicity | To assess NK cell infusion -related toxicity at the bedside. We will monitor the patient and check the vital sign. If any side effect are noticed, we will record and report to the data safety monitoring comittee. | 1 year | 
| Anti-HCC Effect of This Treatment |  | 2 year | 
| Anti-HCV Effect of This Treatment (If Applicable) |  | 2 year |