CAR-NK Project

Final Project Report CAR-NK (download here)

CAR-NK is the first Romanian project that advances the genetic engineering of cells of  the immune system for the treatment of cancer. More specifically, Natural Killer (NK) cells, a particular type of white blood  cells, normally responsible for eliminating emerging tumor cells, are armed with new genes encoding chimeric antigen receptors (CARs) that  specifically bind molecules expressed by the cancerous cells on their surface. CARs are hybrid molecules that comprise different modules taken from different genes: a targeting module, usually a simplified antibody molecule, that binds specifically a molecule on the targeted cell, a transmembrane domain spanning the cellular membrane, and intracellular modules that activate cytotoxic cellular pathways upon binding of the targeting module to the intended target.

As certain types of cancers can express particular targets on their 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, which have been studied for several decades and have recently been approved  commercially for clinical onco-therapies, NK cells have been less studied for this kind of applications, but show promise as an alternative to T cells for the treatment of malignancies.

The overarching aim of the project is to develop in Romania, through in-house and international expertise, research and innovation-development as well as treatment capacities in the field of onco-immunology.

The operational objectives of the project are to develop advanced personalized immunotherapies for various types of cancer based on CAR-NK cells and to create the   first  “off the shelf” human CAR-NK cell bank.

Preliminary results:

  • We have generated effector NK cell populations from peripheral blood mononuclear cells (PBMCs) isolated from healthy donors.
  • We have developed a procedure for increasing the proliferation rate of activated Natural Killer cells.
NK-92 cell line in culture
Primary NK cells isolated from human donors
  • We have transduced activated natural killer cells with anti-EGFR-CAR vectors.
    Overexpression of EGFR, a receptor tyrosine kinase member of ErbB family, has been observed in several kinds of 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. Targeting EGFR might prove beneficial for many cancer patients.

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

  • We have observed high cytotoxic capacity of transduced natural killer cells against EGFR-expressing tumor cell lines derived from breast cancer, MDA-MB468 and SK-BR-3.
Cytolytic activity of NK cells transduced with anti-EGFR-CAR vectors


Pius Branzeu Clinical Emergency Hospital
300723 Timisoara, Romania
156, Liviu Rebreanu Boulevard

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