Acute Myeloid Leukemia, Minimal Residual Disease, Natural Killer Cell
Conditions
Brief summary
This is a interventional, transplantation study. The procedure under study is the infusion of alloreactive NK cells in adult AML patients, eligible for ASCT, who achieved CR after conventional chemotherapy, but harbor MRD-positivity. Haploidentical KIR-L mismatched donors will be included if present at least one allele mismatch at a class I locus among the following ones: HLA-C alleles with Asn77-Lys80, HLA-C alleles with Ser77-Asn80, HLA-Bw4 alleles. KIR-L mismatched donor alloreactive NK cell repertoire will be evaluated in order to determine the functional cell dose to be used for NK cell collection. Phenotypical analysis of KIRs will be correlated to functional tests. NK cells will be selected from a steady-state large volume leukapheresis product from a suitable haploidentical KIR-ligand incompatible donor. NK cell purification will be performed if the donor leukapheresis product contains at least 10x106 NK cells/Kg. Immunomagnetic enrichment of NK cells will follow two subsequent steps: 1) depletion of CD3+ T cells followed by 2) positive selection of CD56+ NK cells. Patients will receive immunosuppressive chemotherapy, fludarabine (Flu) 25 mg/mq/ from day -5 to -3 and cyclophosphamide (Cy) 2 g/mq on day -2 (Flu/Cy). Two days after Cy administration, patients will be infused intravenously with a single dose of cryopreserved NK cells (day 0), which will be followed by subcutaneous administration of IL-2 (10 x 106 IU/day, 3 times weekly) for 2 weeks (6 doses total). PB samples will also be collected for biological studies. In particular, PB samples will be collected for molecular assessment of microchimerism and tracking of haploidentical NK cells for 30 days, immunophenotype studies, alloreactive NK cells cloning and functional assays (cytotoxicity). Enrolled patients will be followed up for at least 12 months after NK cell infusion.
Interventions
BIOLOGICALInfusion of alloreactive NK cells
Infusion of alloreactive NK cells for acute leukemia patients harboring minimal residual disease after conventional chemotherapy and prior to allogeneic stem cell transplantation
Sponsors
IRCCS Azienda Ospedaliero-Universitaria di Bologna
Study design
Allocation
NA
Intervention model
SINGLE_GROUP
Primary purpose
TREATMENT
Masking
NONE
Eligibility
Sex/Gender
ALL
Age
18 Years to No maximum
Healthy volunteers
No
Inclusion criteria
* Diagnosis of de novo or secondary AML * Age ≥ 18 years * Morphologic CR * Eligibility for ASCT * MRD-positivity after induction chemotherapy * Availability of a KIR-L incompatible haploidentical donor * Performance Status ≥ 70% (Karnofsky score) or ≤ 2 (WHO). * Adequate renal (serum creatinine \< 2 mg/dl), pulmonary (Sat O2 ≥ 96%) and hepatic (ALT/AST \< 2.5 x N) function. * Left Ventricular Ejection Fraction (LVEF) of \>50% as determined by Echocardiogram (ECHO). * Signed informed consent.
Exclusion criteria
* Diagnosis of AML FAB M3 * HIV positivity. * HCV positivity with high viral load. * Pregnant or nursing females. * Current uncontrolled infection. * Signs or symptoms of fluid retention (e.g. pleural effusion).
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Safety of infusing a functional target cell dose of 2x105 alloreactive NK cells/kg in AML patients,eligible for ASCT, with persistent MRD after conventional chemotherapy | 48 months | Safety of infusing a functional target cell dose of 2x105 alloreactive NK cells/kg in AML patients,eligible for ASCT, with persistent MRD after conventional (number of adverse events and severe adverse events) |
| Feasibility of collecting a functional target cell dose of 2x105 alloreactive NK cells/kg in at least 70% of patients entering the study | 48 months | Feasibility of collecting a functional target cell dose of 2x105 alloreactive NK cells/kg in at least 70% of patients entering the study as evaluated by in vitro cytotoxicity assay. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Relapse-free survival (RFS) of patients infused with alloreactive NK cells | 48 months | Relapse-free survival (RFS)of patients infused with alloreactive NK cells |
| overall survival (OS) of patients infused with alloreactive NK cells | 48 months | overall survival (OS) of patients infused with alloreactive NK cells |
| Assess the predictive impact of donor NK cell repertoire on overall survival | 48 months | Assess the predictive impact of donor NK cell repertoire as evaluated as the number of alloreactive NK cells/kg, in terms of overall survival |
| Number of patients who achieve and maintain MRD negativity after infusion of alloreactive NK cells | 48 months | Number of patients who achieve and maintain MRD negativity after infusion of alloreactive |
| Evaluate the microchimerism of patients receiving human NK cells | 48 months | Evaluate the microchimerism of patients receiving human NK cells, as evaluated as percent of donor's cells into recipients |
| Functionally evaluate, in vitro and ex vivo, the antitumor activity of infused NK cells | 48 months | Functionally evaluate, in vitro and ex vivo, the antitumor activity of infused NK cells, as evaluated by in vitro cytotoxicity assay (percent of lysis by donor NK cells of recipients cells). |
| Assess the molecular features of infused NK cells | 48 months | Assess the molecular features of infused NK cells, qualitative gene expression profile. |
| Assess the predictive impact of donor NK cell repertoire on relapse-free survival | 48 months | Assess the predictive impact of donor NK cell repertoire as evaluated as the number of alloreactive NK cells/kg, in terms of relapse-free survival |
| Number of patients who enter ASCT in MRD-negativity | 48 months | Number of patients who enter ASCT in MRD-negativity |
Countries
Italy
Contacts
Primary ContactAntonio Curti
Outcome results
None listed