Screening of novel CAR - a step toward local CAR-T Cell production in Pakistan

Chimeric Antigen Receptor T-cell (CAR-T) therapy has transformed outcomes for several malignancies, but prohibitive costs and challenges in transporting viable cells limit access in low- and middle-income countries (LMIC). These barriers contribute to persistently high LMIC mortality even for common, otherwise curable cancers such as B-cell acute lymphoblastic leukemia (B-ALL). To address this disparity, Indus Hospital & Health Network (IHHN) in Karachi, where care is provided free of charge, has initiated foundational work toward developing indigenous CAR-T production capacity.

CAR-T therapy at IHHN, where the pediatric cancer unit registers over 1,200 new cases annually, will require phased development of infrastructure, laboratory workflows, and human capacity. Initial efforts focused on establishing an in-vitro assay to screen CAR constructs by transducing immortalized CD4⁺ T-cells (Jurkat) with lentiviral vectors expressing candidate CARs, followed by co-culture with peripheral blood mononuclear cells (PBMCs) isolated from B-ALL patients. Subsequent phases will extend this workflow to patient-derived CD4⁺ T-cells and, ultimately, to full CAR-T cell manufacturing.

Using Jurkat cells, we demonstrated the ability to evaluate CAR performance through in-vitro assays. We successfully produced lentiviral vectors containing CAR backbones, generated stable CAR-expressing (Jurkat) cell lines, and established the infrastructure and expertise for PBMC isolation. Downstream assessment of CAR efficacy utilized existing platforms, including multicolor flow cytometry. The next phase will involve replicating these findings using primary CD4⁺ T-cells, while the final phase will focus on establishing GMP-compliant CAR-T cell production. Key enabling factors include an existing ISO Class 5 clean room, essential laboratory infrastructure, and institutional commitment to specialized workforce training.

This program provides essential early evidence supporting the feasibility of indigenous CAR-T development in Pakistan. Such building of local expertise and infrastructure will be the only way to create a sustainable, affordable CAR-T production pipeline that directly advances health equity.