The project aims to carry out work essential for the implementation of chimeric antigen receptor T – cell therapy (CAR-T) in Kazakhstan. CAR-T therapy has become one of the most significant advancements in the treatment of hematological malignancies such as leukemia, lymphoma, and multiple myeloma. CAR-T is a therapeutic method based on the use of genetically modified T – lymphocytes that, in addition to their native T – cell receptor, express an artificially created protein, the chimeric antigen receptor (CAR). Project Idea: Produce CAR+ cells from lymphocytes of patients or donors, determine the fractional composition of CAR+ cells (naïve, central memory cells, effector memory cells, terminally differentiated effector cells), and study the influence of the subpopulation composition of CAR+ cells on the potency of the cellular product in in vitro tests. During the production of CAR+ cells, the CliniMACS Prodigy manufacturing system will be launched, and experience in producing and characterizing cellular products will be gained, which will be documented as a prototype for future production guidelines for therapeutic products at the National center for biotechnology.
CAR-T therapy has revolutionized modern oncohematology because it delivers remarkable results in treating leukemia, lymphoma, and multiple myeloma, particularly in clinical forms resistant to traditional therapies (relapsed or refractory disease). For this reason, CAR-T was named “Breakthrough of the Year” in 2013 by the journal Science.
CAR-T therapy is a form of adoptive cell therapy (ACT), a group of biomedical technologies based on the use of immune system cells (either natural or genetically modified) that are extracted from the patient’s body, cultured ex vivo, processed, and then reintroduced into the patient to achieve a therapeutic effect. The general idea of ACT immunotherapy as a group of methods is that, through selected ex vivo culture processing, a large proportion of immune cells can be endowed with new properties. For example, in the case of CAR-T technology, T – lymphocytes can be given a new, uniform immunological specificity and directed to destroy tumors. After reinfusion into the patient’s body, CAR-T lymphocytes target and eliminate cells marked with the antigen against which the CAR receptor is directed.
From the year of the first registration of a CAR-T therapeutic product (2017) to March 2025, eight commercial products with brand names have been registered worldwide: Kymriah (supplier – Novartis, Switzerland), Yescarta (Kite Pharma, USA), Tecartus (Kite Pharma), Breyanzi (Bristol Myers Squibb, USA), Abecma (jointly by BMS and Bluebird Bio, both USA), Carvykti (jointly by Janssen Biotech (USA) and Legend Biotech (China)), Relma-cel (JW Therapeutics, China), and Carteyva (Legend Biotech, China). It is important to note that a significant portion of CAR-T therapies currently used in clinics are produced in non-industrial settings, meaning they are not registered as medicinal products and are used under rules permitting the application of high – tech drugs without clinical trials or registration procedures (so-called “hospital exemption”). For this reason, the annual number of CAR-T procedures performed worldwide, even excluding clinical trials, far exceeds the sales of registered products, with a total of 40,000–50,000 CAR-T procedures conducted globally each year.
As clinical experience accumulates, the list of indications for CAR-T therapy expands, and approaches to treatment sequencing are revised. For example, until the last two years, CAR-T was prescribed only after the failure of all standard therapies (chemotherapy, transplants). However, in 2024, the National Comprehensive Cancer Network (NCCN) published updated lymphoma treatment guidelines recommending CAR-T therapy as a second – line treatment to achieve remission in patients after the failure of standard chemotherapy (i.e., before high – dose chemotherapy and transplants).
CAR-T technology is not yet used in medical institutions in Kazakhstan, but several Kazakh patients have already received CAR-T therapy abroad.
The demand for CAR-T therapy among patients in Kazakhstan is high: approximately 300 new cases of hematological malignancies are diagnosed annually, and (across all etiologies) about 20–30% of these patients eventually face relapse or refractory disease, meaning they require CAR-T therapy.
It is necessary to localize the production and application of CAR-T therapy in Kazakhstan because only local production in an “academic format” can make CAR-T therapy financially accessible to patients in Kazakhstan. Currently, the price of registered CAR-T products dominating the markets of developed countries is extremely high: Kymriah costs $608,000 USD per patient, and Yescarta costs $424,000 USD per patient. The project creates the prerequisites for and is aimed at establishing domestic production of CAR-T therapeutic cell products in Kazakhstan. This should ultimately reduce costs and accelerate access to therapy for patients in advanced stages of the disease, for whom the speed of receiving the product is critical.
Produce CAR+ cells from lymphocytes of patients or donors, determine the fractional composition of CAR+ cells (naïve, central memory cells, effector memory cells, terminally differentiated effector cells), and study the influence of the subpopulation composition of CAR+ cells on the potency of the cellular product in in vitro tests.
Obtain human CAR+ cell preparations from different donors. Collect data on the immunophenotypic variability of CAR+ cells derived from different donors. Assemble a collection of human neoplastic B – cell lines for use in in vitro tests to measure the functional activity of finished CAR-T products. Develop certified methods for production, quality control, and pre – infusion testing of CAR+ cells. Document standard operating procedures (SOPs) and a prototype production guideline. Obtain data on the ability of CAR+ cells to activate, proliferate, persist, and produce cytokines in the presence of CD19+ targets from actual patients. Describe statistical correlations between the content of CAR+ cell subtypes and their activity.
The project has scientific novelty for Kazakhstan due to the absence of domestic production and application of CAR-T products in the country. The project is significant for the development of gene and cell therapy (CGT) technologies in Kazakhstan.
Shustov Alexander Vyacheslavovich, Candidate of Biological Sciences, Project Leader, Head of the Genetic Engineering Laboratory at RSE “National Biotechnology Center.” Research interests: molecular biology, immunology, genetic engineering, cell – based products, gene therapy, and the application of immunotherapy in medicine. Over the past 5 years, he has led 5 projects in the field of molecular biology.
H – index: 11 (Scopus).
ORCID: 0000 – 0001 – 9880 – 9382
Scopus Author ID: 7003352587
ResearcherID: K – 1148 – 2013
Victoria Vladimirovna Keyer, Master of Biology, citizen of the Republic of Kazakhstan, H – index 2, junior researcher at the Genetic Engineering Laboratory, NBC. ResearcherID (Publons): AAU-3714-2020; Scopus Author ID: 57211028385; ORCID: 0000 – 0001 – 8885 – 2387; H – index (Scopus): 2. Role in the project: maintains the cell culture bank, prepares lentiviral vectors, produces cell preparations, operates the Prodigy system, processes bone marrow samples, obtains neoplastic cell lines, and performs immunophenotyping of B – cell lines.
Laura Rizabekovna Syzdykova, Master of Engineering and Technology, PhD candidate at L.N. Gumilyov Eurasian National University (current doctoral studies 2022–2025), citizen of the Republic of Kazakhstan, H – index 2 (Thomson Reuters), H:2 (RSCI), junior researcher at the Genetic Engineering Laboratory, NBC. ResearcherID: DXQ – 3837 – 2022; ORCID: 0000 – 0002 – 1785 – 6978; Scopus Author ID: 57221476753; H – index (Scopus): 1. Role in the project: produces CAR-T cell preparations, operates the Prodigy system, immunophenotypes CAR-T cell subpopulations, and conducts proliferation and cytotoxicity assays.
Gulzat Maratovna Zauatbayeva, Master of Technical Sciences with a specialization in biotechnology, junior researcher at the Genetic Engineering Laboratory, NBC. Role in the project: maintains the cell culture bank, isolates plasmids, and performs cloning.
In this study, for the first time in Kazakhstan, cellular products containing CAR-T cells have been produced. The preparations were manufactured using the CliniMACS Prodigy system with technologies from Miltenyi Biotec (a global leader in technological solutions for localized cell therapy production), marking the successful transfer of CAR-T technology to Kazakhstan. The CAR-T cell preparations were derived from apheresis products of donors and patients. Two different CAR receptor constructs were used, both targeting the B – cell/blast antigen CD19 (based on the anti – CD19 scFv FMC63). One of the CAR receptors is molecularly analogous to the structure published for Kymriah therapy, while the other is analogous to Yescarta therapy. The CAR-T preparations produced at the National Biotechnology Center meet the technical specifications for absolute CAR+ cell content, functional activity of CAR+ cells, and safety for infusion (sterility), and are comparable to registered products used globally for CAR-T therapy. Data on the immunophenotypic variability of CAR+ cells derived from different donors have been collected. A prototype production guideline for CAR-T therapeutic cell products is under development.