Applied Genome Editing for Advanced Therapies

Cancer immunotherapy represents a revolutionary approach harnessing the body’s immune system to eradicate tumor cells. One particularly promising avenue is adoptive cell therapy (ACT), notably CAR-modified T cells, which has revolutionized the landscape of immunotherapies and enhanced patient outcomes. However, despite significant strides in CAR-T therapies for certain hematologic diseases, their effectiveness in treating more challenging conditions such as acute myeloid leukemia (AML) and solid tumors remains limited. Due to high immunosuppression of the diseases and the difficulty to obtain enough autologous T cells. To confront these challenges, the research group “Applied Genome Editing for Advanced Therapies (eGATA),” under the leadership of Dr. Karim Benabdellah, aim to increase the efficacy of allogenic CAR-T cell therapy through a multifaceted approach. The group’s robust international collaborations, spearheaded by Dr. Benabdellah’s leadership in the COST action, and close collaboration with Dr. Herrera at Reina Sofia Hospital, serve as pillars for its purpose. Based on this foundation, eGATA encompasses 5 complementary research lines:

Enhancement of Tumor Microenvironment Targeting Strategy

This research line aims to improve the efficacy of CAR T-cell therapy, particularly in pathologies where these cellular products are not yielding comparable results to B-lineage neoplasms, such as AML, MM, and solid tumors. Understanding the cellular and humoral mechanisms governing the TME and its strongly immunosuppressive effects on CAR-T activity is crucial. We are developing combined strategies utilizing genetic engineering and nanoparticles (primarily exosomes and also LNPs) to enhance CAR T-cell therapy effectiveness.

Preclinical Development of Universal EXO-CART and Their Potential Application in Cancer Immunotherapy Protocols

Through this innovative research line, we seek to implement a therapeutic exosome production platform to complement CAR therapy in situations where CAR-T cells alone are ineffective, such as solid tumors and some leukemias. We are improving exosome production in CAR-T cells through genomic manipulation. Our group possesses a range of CRISPR/Cas9-based systems capable of efficiently editing the genome of primary T lymphocytes and various viral systems for overexpression of therapeutic proteins to enhance exosome production efficiency.

Enhancement of Anti-CD19 CAR-T Cells via Controlled Expression of Factors and Cytokines (Fourth-Generation CAR-T Strategy)

In this research line, we are developing CAR-T cells, by the way of gene editing approaches, with additional functionalities by controlling the expression of cytokines with potential to improve efficacy and thus the anti-tumor activity of CAR-T cells. Controlled secretion of these cytokines could enhance the phenotype and potential of edited cells, aiming to improve the efficacy of CAR-T-based therapies. We are developing a PD1-based platform to allow for the expression of cytokine such as IL15 genes, as a proof of concept, to increasing the efficacy, and specificity of fourth-generation CAR-T cells for eliminating CD19+ B neoplasms.

Exploration of New Sources of T Lymphocytes

n another line of research, we are exploring the possibility of generating allogeneic CAR-T cells from alternative cellular sources such as umbilical cord blood (CB-CD19-CAR-T) and NK cells from various sources (NK-CAR-CD19). We aim to evaluate the potential advantages and disadvantages of different approaches to the use of allogeneic CARs (TCR and/or HLA KO peripheral blood CAR-T cells, NK-CAR, and CB-CAR T cells).

Universal CAR-T Immunotherapy Based on Subpopulation Selection and Generation of Universal TCR and HLA-/- CAR-T Cells

Over the past four years, we have generated universal CAR-T cells by eliminating TCR and HLA class II. We have also selected less differentiated universal CAR-T cell populations (TCR KO and HLA KO), thereby enhancing the universal product’s persistence. Our current program aims to improve large-scale cell cultures to expand and freeze double KO universal cells with a well-defined phenotype, facilitating the initiation of a clinical trial in Andalusia with CD19-CART universal DKO T cells.