CD19 CAR-expressing iPSC-derived NK cells effectively enhance migration and cytotoxicity into glioblastoma by targeting to the pericytes in tumor microenvironment.

In cancer immunotherapy, chimeric antigen receptors (CARs) targeting specific antigens have become a powerful tool for cell-based therapy. CAR-natural killer (NK) cells offer selective anticancer lysis with reduced off-tumor toxicity compared to CAR-T cells, which is beneficial in the heterogeneous milieu of solid tumors. In the tumor microenvironment (TME) of glioblastoma (GBM), pericytes not only support tumor growth but also contribute to immune evasion, underscoring their potential as therapeutic targets in GBM treatment. Given this context, our study aimed to target the GBM TME, with a special focus on pericytes expressing CD19, to evaluate the potential effectiveness of CD19 CAR-iNK cells against GBM. We performed CD19 CAR transduction in induced pluripotent stem cell-derived NK (iNK) cells. To determine whether CD19 CAR targets the TME pericytes in GBM, we developed GBM-blood vessel assembloids (GBVA) by fusing GBM spheroids with blood vessel organoids. When co-cultured with GBVA, CD19 CAR-iNK cells migrated towards the pericytes surrounding the GBM. Using a microfluidic chip, we demonstrated CD19 CAR-iNK cells' targeted action and cytotoxic effects in a perfusion-like environment. GBVA xenografts recapitulated the TME including human CD19-positive pericytes, thereby enabling the application of an in vivo model for validating the efficacy of CD19 CAR-iNK cells against GBM. Compared to GBM spheroids, the presence of pericytes significantly enhanced CD19 CAR-iNK cell migration towards GBM and reduced proliferation. These results underline the efficacy of CD19 CAR-iNK cells in targeting pericytes within the GBM TME, suggesting their potential therapeutic value for GBM treatment.

Agreement of New Automated Matched Alternation Flicker using Undilated Fundus Photography for the Detection of Glaucomatous Structural Change.

PURPOSE: To determine the agreement among glaucoma experts and general ophthalmologists regarding detection of glaucomatous structural changes using a new automated matched alternation flicker (AMAF) method with fundus photographs (FPs) of undilated eyes. METHODS: Sixty-six pairs of FPs of normal tension glaucoma patients were collected. FPs were taken at intervals of more than 12 months. Alternating flicker images were created using a new AMAF application. In a blinded manner, two glaucoma experts and two general ophthalmologists compared the presence of glaucomatous structural changes using either the AMAF method or the side-by-side comparison method. The interobserver and intraobserver agreements were compared using the Bland and Altman plot analysis. RESULTS: The glaucoma experts detected more glaucoma progression using the AMAF method (average, 50.7%) compared with the side-by-side method (average, 32.5%). General ophthalmologists detected more glaucomatous progression with the AMAF method (average, 40.9%) than with the side-by-side method (average, 25.0%). The AMAF method showed fair to substantial interreader agreement (k = 0.511-0.724) and fair to perfect intrareader agreement (k = 0.631-0.943). Interreader and intrareader agreements using the AMAF method were better for the glaucoma experts compared with the general ophthalmologists. CONCLUSIONS: The AMAF method showed more changes in FPs than the classical side-by-side comparison method. Regarding inter- and intrareader agreements, agreement for the glaucoma experts was best using the AMAF method, but for the general ophthalmologists agreement was best using the side-by-side comparison method.