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

The National Cancer Institute awarded the University of Miami $110,228 on December 1, 2026, under the Cancer Research Manpower program (CFDA 93.398) to develop and characterize chimeric ligand-presenting T cells (CLIP-T cells) for treating extranodal NK/T-cell lymphoma, a rare and aggressive subtype of non-Hodgkin's lymphoma.

The project, performed in Miami, Florida through May 31, 2030, focuses on engineering CLIP-T cells expressing stress-induced ligands (HLA-E or MICA) to bind ENKTL-specific receptors (NKG2A or NKG2D), with parallel development of conventional CAR-T cells bearing single-chain variable fragments against the same targets for comparative functional analysis. The central hypothesis posits that CLIP-T cells leveraging ligand-receptor interactions will prove more effective against ENKTL than receptor-receptor interactions of scFvs in traditional CAR-T cells, based on the innate binding affinity of endogenous proteins and synergistic triggering of target receptor biological effects. The funded work encompasses two specific aims: in vitro characterization and refinement of HLA-E CLIP-T, MICA CLIP-T, anti-NKG2A CAR-T, and anti-NKG2D CAR-T cells, and comparative in vivo tolerance and efficacy studies. This addresses an unmet clinical need; while CAR-T therapy has transformed treatment of relapsed or refractory B-cell NHL, no cellular therapy has been developed specifically for ENKTL, which carries a five-year survival rate below 30% in advanced or refractory cases.

Abstract

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.