Surface Engineering of Natural Killer Cells with CD44‐targeting Ligands for Augmented Cancer Immunotherapy

Abstract

Adoptive immunotherapy utilizing natural killer (NK) cells has demonstrated remarkable efficacy in treating hematologic malignancies. However, its clinical intervention for solid tumors is hindered by the limited expression of tumor‐specific antigens. Herein, lipid‐PEG conjugated hyaluronic acid (HA) materials (HA‐PEG‐Lipid) for the simple ex‐vivo surface coating of NK cells is developed for 1) lipid‐mediated cellular membrane anchoring via hydrophobic interaction and thereby 2) sufficient presentation of the CD44 ligand (i.e., HA) onto NK cells for cancer targeting, without the need for genetic manipulation. Membrane‐engineered NK cells can selectively recognize CD44‐overexpressing cancer cells through HA‐CD44 affinity and subsequently induce in situ activation of NK cells for cancer elimination. Therefore, the surface‐engineered NK cells using HA‐PEG‐Lipid (HANK cells) establish an immune synapse with CD44‐overexpressing MIA PaCa‐2 pancreatic cancer cells, triggering the “recognition‐activation” mechanism, and ultimately eliminating cancer cells. Moreover, in mouse xenograft tumor models, administrated HANK cells demonstrate significant infiltration into solid tumors, resulting in tumor apoptosis/necrosis and effective suppression of tumor progression and metastasis, as compared to NK cells and gemcitabine. Taken together, the HA‐PEG‐Lipid biomaterials expedite the treatment of solid tumors by facilitating a sequential recognition‐activation mechanism of surface‐engineered HANK cells, suggesting a promising approach for NK cell‐mediated immunotherapy.

Optimized Design of Hyaluronic Acid–Lipid Conjugate Biomaterial for Augmenting CD44 Recognition of Surface-Engineered NK Cells

“…Similarly, in our previous study, HA functional group-coated NK cells were tested with the CD44 blocking MDA-MB-231 model. The results showed comparable efficacy to uncoated NK cells when treated with MDA-MB-231 . This outcome provides evidence that the HA component of our coating material targets CD44 on cancer cell surfaces.…”

“…The results showed comparable efficacy to uncoated NK cells when treated with MDA-MB-231. 38 This outcome provides evidence that the HA component of our coating material targets CD44 on cancer cell surfaces. While HA moieties can enhance recognition and targetability toward CD44-overexpressing cancers, the facile incorporation of PEG and lipid into NK cell membranes allows for simple ex vivo cell surface modification.…”

Networked Cluster Formation via Trigonal Lipid Modules for Augmented Ex Vivo NK Cell Priming

“…This approach enables the augmented NK cell activation without the requirement of additional supplementary cytokine cocktails. Our results demonstrated that (1) TL modules could be effectively immobilized into the lipid bilayers of NK cell membranes, specifically via hydrophobic interaction [33,34], and (2) they could strengthen the membrane connections of NK cells through material-mediated physical links. In consequence, as compared with naturally-driven NK clustering, which is mediated by ECM production (without the aid of biomaterials), a significant induction of multiple membrane interactions occurred, leading to rapid and tight formation of NK cell clusters, and effective cell priming ultimately took place, resulting in enhanced IFN-γ secretion in TLNK spheroids.…”

Targeted Ferroptosis-Immunotherapy Synergy: Enhanced Antiglioma Efficacy with Hybrid Nanovesicles Comprising NK Cell-Derived Exosomes and RSL3-Loaded Liposomes

“…It has been proposed that apoptosis is an important mechanism in NK cell-based immunotherapy . Perforin triggers the intrinsic pathway and elevates the release of cytochrome-c . FASL triggers the extrinsic apoptotic pathway by activating caspase-3. , To confirm the cell death mechanism, we examined the expression of apoptosis-related proteins (caspase-3 and cytochrome-c) using IHC staining.…”