Engineering Chimeric Ligand Presenting T Cells (CLiP-Ts) Targeting Receptors of Extranodal NK/T-Cell Lymphoma: A Comparative Study with Traditional CAR-T Cell Therapy

Extranodal NK/T cell lymphoma (ENKTL) is a rare, aggressive subtype of non-Hodgkin’s lymphoma (NHL). It is characterized by extra-nodal tumor growth, particularly in the nasal cavity, skin, and gastrointestinal tract. ENKTL accounts for up to 15% of new lymphoma cases in specific populations. A majority (85%) of ENKTL cases originate from natural killer (NK) cells. ENKTL remains a therapeutic challenge, with advanced or refractory cases carrying a five-year survival rate < 30%. While chimeric antigen receptor T cell (CAR-T) therapy has revolutionized treatment of relapsed or refractory B-cell NHL, no cellular therapy has been developed specifically for ENKTL. The overall objective of this project is to develop a novel class of chimeric ligand-presenting (CLiP) T cells for ENKTL treatment. Each CLiP-T cell expresses a stress-induced ligand, HLA-E or MICA, on its surface to bind ENKTL-specific receptors, NKG2A or NKG2D, respectively. Concurrently, conventional CAR-T cells with a single-chain variable fragment (scFv) against NKG2A or NKG2D were created for comparison in all functional studies. The central novel hypothesis is that CLiP-T cells leveraging ligand-receptor interactions are more effective in treating ENKTL than receptor-receptor interactions of scFvs in CAR-T cells. The rationale is that the ligand-based approach leverages the innate ability of endogenous proteins to bind tightly to their target receptors, thereby directing the antitumor activity of the CLiP-T cell while simultaneously triggering the target receptor’s biological effects, creating a synergistic interaction. The central hypothesis will be tested through two specific aims: 1) Characterization and refinement of HLA-E CLiP-T, MICA CLiP-T, anti-NKG2A CAR-T, and anti- NKG2D CAR-T in vitro, and 2) Compare in vivo tolerability, persistence, and efficacy against ENKTL tumors—HLA-E CLiP-T vs. anti-NKG2A CAR-T and MICA CLiP-T vs. anti-NKG2D CAR-T. The goal of Aim 1 is to understand the biology of CLiP-T cells in comparison to CAR-T cells and to refine three supplementary design components. Aim 2 focuses on comparing the functionality of the CLiP-T and CAR-T cells in vivo. Rigorous, reproducible preliminary data support this novel hypothesis-driven proposal: we validated all CLiP-T and CAR-T constructs and demonstrated that CLiP-T cells eliminate NK92 cells in vitro more effectively than their CAR-T counterparts. The expected outcomes of this proposal include the development of two ligand- based CLiP-T cells expressing HLA-E or MICA as potential disease-specific cellular therapies for ENKTL. These studies are significant because they will establish a first-in-class, disease-specific cellular therapy for ENKTL, provide key insights into NK-cell immune regulation, and offer the first true comparison of ligand- and scFv-based CAR-T approaches. These innovative studies evaluate multiple previously untargeted receptors for cellular therapy and will be the first application of stress-induced proteins to direct CAR-T cell antitumor activity. This work has broad applicability; CLiP-T technology allow virtually any receptor to be targeted therapeutically. This proposal offers exceptional training in the design, construction, and evaluation of cellular therapies. Project Number: 1F30CA310065-01 | Fiscal Year: 2026 | NIH Institute/Center: National Cancer Institute (NCI) | Principal Investigator: Daniel Tsai | Institution: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE, CORAL GABLES, FL | Award Amount: $55,114 | Activity Code: F30 | Study Section: Special Emphasis Panel[ZRG1 F09C-H (22)] View on NIH RePORTER: https://reporter.nih.gov/project-details/11316335