Department of Fundamental Oncology with Genetic Engineering and Cell Therapy Group

Alexei Petukhov
Head of Group

Principal activity

The Genetic Engineering and Cell Therapy Group focuses on research of the efficacy and safety of genetically modified lymphocytes for cancer treatment. The laboratory is equipped with bioreactors used for scaling up the production of vectors for genetic modification of T cells, the equipment for the production of cell products, their analysis and cryopreservation.

Newly developed and implemented technologies

In order to determine a strategy for improving the design of CAR-T cells, the Research Laboratory of Onco-Hematology is currently working on the identification of genes associated with tumor resistance to CAR-T therapy. Another area is the development of allogeneic universal CAR-T cells to increase the availability of therapy for patients. The Genetic Engineering and Cell Therapy Group has been carrying out the work on the domestic development of CAR-T cells.

Participation in State Assignments 

Experimental research «Development of allogeneic tumor-specific T cells with an anti-CD19 chimeric antigen receptor» (State Assignment of the Russian Health Ministry).

Supervised by: Prof. Andrey Zaritskey.

Grants

• «Study of the efficacy mechanisms of T cells with chimeric antigen receptor (CAR-T) in cell-based and animal models of solid tumors» (ongoing project).
• “Pathophysiological significance of the PPM1D gene and its therapeutic modulation in a xenograft model of human acute myeloid leukemia” (ongoing project).
• “Reprogramming of myeloid suppressor cells by genomic editing to increase the effectiveness of antitumor immunotherapy (ongoing project).
• “Works in the area of diagnosing significant mutations of acute myeloid leukemia (IDH, FLT3, DNMT3a, etc.) ” (ongoing project).
• Tumor resistance genes for MHC-independent immunotherapy (completed in 2019).

Major results: CAR-T therapy is effective in treating hematologic malignancies, but the chance of relapse may still be high. Our objective was to identify genes that help tumor cells suppress the effectiveness of CAR-T cell therapy or to identify genes that can alter the expression profile in tumor cells after CAR-T exposure. To identify such resistance genes, we performed genomic screening with the CRISPR / Cas9 Synergistic Activation Mediator (SAM) library and next generation sequencing. We used Hela cells that exogenously express CD19 as a model system. The studies have identified immunosuppressive genes that may be associated with the effectiveness of CAR-T therapy. CD19 + Hela cells were incubated with CAR-T cells (anti-CD19 with 4-1BB or CD28). The surviving clones were sequenced. The results were validated by screening in several rounds of library enrichment (surviving clones of tumor cells were again incubated with CAR-T cells) and further bioinformatic analysis, as well as by overexpression of candidate genes in the Hela tumor line followed by incubation with CAR-T cells. The results can be used to boost the effectiveness of CAR-T therapy.

International collaboration 

The Genetic Engineering and Cell Therapy Group collaborates with the French National Institute of Health and Medical Research (INSERM), the Bourgogne Franche Comté University (France); the University of Texas MD Anderson Cancer Center (USA).