Abstract B009: Developing a platform for CEA-directed CAR-NK cells expressing IL-15 and CCL21 for the treatment of non-small cell lung cancer
Authors: Rennels Austin K , Lee Ji-Ann , Abascal Jensen , Crosson William P , Kim Faith , Oh Michael , Kahangi Bitta P , Reyes Edgar Perez , Coleman Nalani J , Dumitras Camelia , Bottero Carlos , Jiang Hong , Krysan Kostyantyn , Tran Linh M , Salehi-Rad Ramin , Liu Bin , Dubinett Steven M 
Published: February 2025

Lung cancer remains the leading cause of cancer death in the U.S. and worldwide. Non-small cell lung cancer (NSCLC) affects 85% of patients with lung cancer, of which only 17% will survive beyond 5 years post-diagnosis. Immune checkpoint blockade (ICB) therapies, including anti-PD-1/PD-L1, have become standard-of-care treatments for NSCLC. However, ICB monotherapy is ineffective in most cases due to mechanisms of either de novo or acquired resistance, resulting in patients who either do not respond to treatment or others who relapse after an initial response. Thus, there is a critical need to develop innovative cellular therapeutics for lung cancer to overcome the limitations of current targeted immunotherapies. Recent advancements in cellular immunotherapies have significantly transformed the landscape of cancer treatment. Notably, the success of chimeric antigen receptor- (CAR) engineered T cell therapy for hematological malignancies underscores the significance of these developments. However, despite their success in blood cancers, adoptive cell therapies (ACT) have shown limited efficacy in solid tumors due to many factors, including tumor antigen heterogeneity, poor trafficking to the tumor site, and the immunosuppressive tumor microenvironment (TME). In addition to the biological limitations, ACT presents logistical challenges such as high manufacturing costs, complex autologous platforms, and potential severe toxicities (e.g., graft-versus-host disease (GVHD)). To address these challenges, we propose the development of a platform for "off-the-shelf" CAR-engineered cell therapy derived from healthy donor peripheral blood NK cells (PB-NK). Allogeneic CAR-NK cells offer advantages, including reduced risk of GVHD, cost-effectiveness, and simplicity of production compared to autologous cell therapy approaches. However, CAR-NK therapies are not without their own pitfalls, such as limited in vivo persistence and lack of systemic immune response. We intend to overcome these limitations by incorporating key elements in our NK cell engineering strategy: 1) integration of carcinoembryonic antigen (CEA)-directed CAR for tumor-specific targeting, 2) incorporation of human IL-15 to promote NK cell survival and expansion, 3) secretion of C-C Motif Chemokine Ligand 21 (CCL21) to promote T cell infiltration and systemic anti-tumor responses, and 4) intratumoral delivery of the cell product to overcome poor in vivo trafficking to the TME and minimize potential toxicities. We hypothesize that the combination of direct tumor killing by engineered CAR-NK cells with host anti-tumor immune responses induced by CCL21 will yield both immediate tumor reduction and long-term immune-mediated tumor control. Our results highlight a feeder-free expansion protocol that allows for high transduction efficiency of PB-NK cells with VSVG-pseudotyped lentivirus encoding for CAR and multi-cytokine cargoes.

Citation Format: Austin K Rennels, Ji-Ann Lee, Jensen Abascal, William P Crosson, Faith Kim, Michael Oh, Bitta P Kahangi, Edgar Perez Reyes, Nalani J Coleman, Camelia Dumitras, Carlos Bottero, Hong Jiang, Kostyantyn Krysan, Linh M Tran, Ramin Salehi-Rad, Bin Liu, Steven M Dubinett. {Abstract title} [abstract]. In: Proceedings of the AACR IO Conference: Discovery and Innovation in Cancer Immunology: Revolutionizing Treatment through Immunotherapy; 2025 Feb 23-26; Los Angeles, CA. Philadelphia (PA): AACR; Cancer Immunol Res 2025;13(2 Suppl):Abstract nr B009.