CAR-NK Cells Production and Expansion From Patients With Lympho or Myeloproliferative Disorders and From Healthy Donors

December 6, 2024 updated by: European Institute of Oncology

Production and Expansion of Chimeric Antigen Receptor (CAR)-Natural Killer (NK) Cells From Patients Affected by Lympho or Myeloproliferative Disorders and From Healthy Donors

Recent studies demonstrated the safety and efficacy of allogeneic and autologous infusion of NK cells as adoptive immunotherapy in malignant hematological diseases and solid tumors.NK cells are innate immunity effectors with antitumor activity regulated by a wide variety of receptors located on their cell surface, with both activating and inhibiting roles.Receptors with an inhibitory role include receptors of the KIRs family (Killer Immunoglobulin like Receptors) and receptors of the CD94 / NKG2A complex and are capable of recognizing molecules of Human Leukocyte Antigen class I (HLA-I). Receptors of the KIRs family are members of the immunoglobulin superfamily and are encoded by highly polymorphic genes located on chromosome 19q13.4 in a region known as a leukocyte receptor cluster (LCR). The predominant ligand for KIRs receptors is HLA-C, but other studies show that HLA-A and HLA-B are also involved. Receptors with an activating role, on the other hand, such as NCRs (Natural Cytotoxic receptors), are specific to NK cells, while other epitopes such as CD 56 are also present on other populations of T lymphocytes.

This receptor condition allows, in patients suffering from haemopathy and undergoing transplantation allogeneic stem cell, an alloreactivity induced by the mismatch between the donor's KIRs and their ligands on recipient target cells and, therefore, the role of NK cells as allogeneic effectors. The engraftment of NK cells has been shown to be correlated with a lower risk of disease recurrence, therefore the therapeutic infusion of NK cells from donors could allow, with benefit, the acquisition of fully functional NK cells in the recipient.

Study Overview

Status

Recruiting

Conditions

Intervention / Treatment

Detailed Description

The primary aim of the study is to manufacture and expand NK cells expressing CD19, CD79 and CD123 CARs from Peripheral Blood Mononucleated Cells (PBMCs) (cohort a) and from CD34+ cells (cohort b and c) of subjects affected by myelo or lymphoproliferative disorders or healthy donors.

The secondary aim is to evaluate their cytotoxic activity in preclinical models. The main steps of CAR-NK cell manufacturing and preparation are the followings

  1. Collection of peripheral venous blood (40 mL collected in EDTA tubes for cohort a, or 10 mL from patients with >20 CD34+ cells/mcL for cohort b), or blood from bone marrow aspiration (5 mL of bone marrow collected in one EDTA tube for cohort b), or a part of LP (2 mL of PL in one EDTA tube for cohort c) from subjects with myelo/lymphoproliferative diseases and from Healthy Donors;
  2. Selection of mononucleated cells through a density gradient centrifugation technique (Ficoll-Histopaque) (cohort a and b);
  3. Selection of CD34 + Hematopoietic stem cells (HSCs) through magnetic activated cell sorting (cohort b)
  4. Expansion NK cells from PBMCs (cohort a), using supplements such as recombinant cytokines, stimulatory antibodies, medicinal products and irradiated cells to support the process;
  5. Expansion of CD34+ cells (cohort b and c), using supplements such as recombinant cytokines, stimulatory antibodies, medicinal products and irradiated cells to support the process, in order to promote differentiation to the NK cell lineage;
  6. Engineering of NK cells through retroviral or lentiviral vectors (VL) that enable transduction with chimeric antigen receptors (CARs) specific for neoplastic cells.
  7. Increasing the activation of NK cells through the down regulation of their inhibitory receptors (CRISPR Cas-9 technology and pharmacological modulation) or pharmacologic modulation of epigenetic, metabolic (HIF1-alpha) and immune-related pathways;
  8. Testing, in in vitro and in vivo preclinical models, the persistence and the activity of the CAR-NK cells produced.

100 patients and 40 healthy donors will be prospectively enrolled over 3 years in order to perform 70 experiments of NK-cell expansion from PBMCs and 70 experiments from CD34+ cells (from PB or bone marrow or LP).

Study Type

Interventional

Enrollment (Estimated)

140

Phase

  • Not Applicable

Contacts and Locations

This section provides the contact details for those conducting the study, and information on where this study is being conducted.

Study Contact

Study Contact Backup

Study Locations

  • Italy
      • Milano, Italy, 20141
        • Recruiting
        • Istituto Europeo di Oncologia
        • Contact:

Participation Criteria

Researchers look for people who fit a certain description, called eligibility criteria. Some examples of these criteria are a person's general health condition or prior treatments.

Eligibility Criteria

Ages Eligible for Study

  • Adult
  • Older Adult

Accepts Healthy Volunteers

Yes

Description

Inclusion Criteria:

  • Subjects over the age of 18 capable of understanding and willing
  • Subjects affected by myelo/lymphoproliferative diseases

Exclusion Criteria:

  • Patients with psychiatric, addictive, or any disorder, which compromises ability to give informed consent for participation in this study.
  • Patients affected by lympho or myeloproliferative disorders and Healthy Donors with known acute and chronic viral and bacterial infections :HIV, HCV (Hepatitis C Virus), HBV (Hepatitis B Virus), syphilis

Study Plan

This section provides details of the study plan, including how the study is designed and what the study is measuring.

How is the study designed?

Design Details

  • Primary Purpose: Basic Science
  • Allocation: Non-Randomized
  • Interventional Model: Parallel Assignment
  • Masking: None (Open Label)

Arms and Interventions

Participant Group / Arm
Intervention / Treatment
Experimental: Cohort a
The collection of PBMCs will be performed during standard blood test procedures: 40 mL of PB will be collected in 8 EDTA tubes.
Procedure: peripheral blood collection
standard blood test procedures
Experimental: Cohort b

The collection of CD34+ cells will be performed during HSC mobilization and staging bone marrow aspirations, as follows:

  1. During standard HSC mobilization, 10 mL peripheral blood will be collected in two EDTA tubes from patients with >20 CD34+ cells/mcL.
  2. 5 mL of bone marrow will be collected in one EDTA tube during bone marrow aspiration performed in the context of standard diagnostic procedures.
Procedure: peripheral blood collection
standard blood test procedures
Experimental: Cohort c
The collection of CD34+ cells will be performed from a LP required in the context of standard therapeutic procedures
Procedure: leukapheresis
Leukapheresis is the procedure that allows the separation and collection of hematopoietic stem cells (HSCs) from peripheral blood. The collection takes place through a machine called a "cell separator": it uses centrifugal force to separate the mononuclear cells from all the other cells in the blood and plasma.

What is the study measuring?

Primary Outcome Measures

Outcome Measure
Measure Description
Time Frame
Primary objective
Time Frame: 3 years
100 patients and 40 healthy donors will be prospectively enrolled over 3 years in order to perform 70 experiments of NK-cell expansion from PBMCs and 70 experiments from CD34+ cells (from PB or bone marrow or LP). We aim to demonstrate feasibility of the program (as defined in the primary endpoint) in 80% of eligible patients in the cohorts a), b) and c). The study will be powered to estimate the fraction of enrolled patients that obtain a full NK expansion defined as A) ≥40 fold ex-vivo expansion of the NK cell population at day 15 of cell culture and B) ≥1x10^9 NK-cells following ex-vivo expansion of 1x10^6 CD34+ cells at day 30 of cell culture; a minimum of 70 patients are required to estimate a true prevalence rate between 70 and 90% with 95% confidence interval (with 56 expected successful cases)
3 years

Collaborators and Investigators

This is where you will find people and organizations involved with this study.

Sponsor

European Institute of Oncology

Investigators

  • Principal Investigator: Enrico Derenzini, MD, Istituto Europeo di Oncologia

Study record dates

These dates track the progress of study record and summary results submissions to ClinicalTrials.gov. Study records and reported results are reviewed by the National Library of Medicine (NLM) to make sure they meet specific quality control standards before being posted on the public website.

Study Major Dates

Study Start (Actual)

December 21, 2022

Primary Completion (Estimated)

December 21, 2025

Study Completion (Estimated)

December 21, 2025

Study Registration Dates

First Submitted

March 19, 2024

First Submitted That Met QC Criteria

December 6, 2024

First Posted (Estimated)

December 10, 2024

Study Record Updates

Last Update Posted (Estimated)

December 10, 2024

Last Update Submitted That Met QC Criteria

December 6, 2024

Last Verified

March 1, 2024

More Information

Terms related to this study

Keywords

Additional Relevant MeSH Terms

Other Study ID Numbers

  • IEO 1886

Plan for Individual participant data (IPD)

Plan to Share Individual Participant Data (IPD)?

NO

Drug and device information, study documents

Studies a U.S. FDA-regulated drug product

No

Studies a U.S. FDA-regulated device product

No

This information was retrieved directly from the website clinicaltrials.gov without any changes. If you have any requests to change, remove or update your study details, please contact register@clinicaltrials.gov. As soon as a change is implemented on clinicaltrials.gov, this will be updated automatically on our website as well.

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