The CAR-NK project

// Project

Project CAR-NK

Duration of the project

51 months

Financing contract

 32 of 01.09.2016


OncoGen-Pius Brânzeu Emergency County Clinical Hospital, Timișoara

Project details


Project summary

CAR-NK is the first Romanian project that advances the genetic engineering of immune system cells for cancer treatment. Specifically, Natural Killer (NK) cells, a type of white blood cell normally responsible for eliminating early-stage tumor cells, are armed with new genes that encode chimeric antigen receptors (CARs), receptors that specifically bind to molecules expressed on the surface of malignant cells. Chimeric antigen receptors are hybrid molecules containing various modules obtained from different genes: a targeting module, usually represented by a simplified antibody molecule that specifically binds a molecule on the target cell, a transmembrane domain that crosses the cell membrane, and intracellular modules that activate cytotoxic cellular pathways upon binding of the targeting module to its intended target.

Given that some types of cancer can express specific targets on the cell surface, immune cells such as NK or T cells can be modified to recognize and attack a specific type of malignant cell.

Unlike engineered T cells, cells that have been studied for decades and have recently been approved for commercial use as clinical onco-therapies, NK cells have been less studied for this type of use, yet represent an alternative promising in T cells for the treatment of malignancies.


The overall objective of this project is the development, in Romania, using local and international expertise, both of the innovative research and development activity, as well as of the treatment possibilities in the field of onco-immunology.

Operational objectives of the project are the development of advanced and personalized immunotherapies for various types of cancer, therapies based on CAR-NK cells, and the creation of the first "off the shelf" human CAR-NK cell bank.

Preliminary results

  • We generated populations of effector NK cells from peripheral blood mononuclear cells (PBMCs) isolated from healthy donors.
  • We have developed a procedure to increase the proliferation rate of activated Natural Killer cells.

NK-92 cell line in culture medium

Primary NK cells isolated from human donors
    • We performed the transduction of activated natural killer cells using anti-EGFR-CAR vectors.
      Overexpression of EGFR, a tyrosine kinase member receptor of the ErbB class of proteins, has been observed in several human malignancies such as non-small cell lung cancer, breast cancer, prostate cancer, gliomas, colorectal cancer or squamous cell carcinomas of the head and neck. Using EGFR as a target could prove beneficial for many cancer patients.

    Video recording showing NK cells recognizing and binding to MCF7 breast cancer cells stained with green fluorescent protein (GFP).

    • We observed that the transduced natural killer cells show an increased cytotoxic capacity against EGFR-expressing cell lines derived from breast cancer, MDA-MB468 and SK-BR-3.
    Activitatea citolitică a celuelor NK transduse cu vectori anti-EGFR-CAR

    Cytolytic activity of NK cells transduced with anti-EGFR-CAR vectors