Pd14-09 High and Selective Cytotoxicity of Ex Vivo Expanded Allogeneic Human Natural Killer Cells From Peripheral Blood Against Bladder Cancer: Implications for Natural Killer Cell Instillation After Transurethral Resection of Bladder Tumor

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology II (PD14)1 May 2024PD14-09 HIGH AND SELECTIVE CYTOTOXICITY OF EX VIVO EXPANDED ALLOGENEIC HUMAN NATURAL KILLER CELLS FROM PERIPHERAL BLOOD AGAINST BLADDER CANCER: IMPLICATIONS FOR NATURAL KILLER CELL INSTILLATION AFTER TRANSURETHRAL RESECTION OF BLADDER TUMOR Fangming Wang Fangming WangFangming Wang View All Author Informationhttps://doi.org/10.1097/01.JU.0001009472.76470.8c.09AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Non-muscle-invasive bladder cancer (NMIBC) is treated with transurethral resection of bladder tumor (TURBT) followed by an intravesical instillation of chemotherapy or Bacillus Calmette-Guérin therapy. However, these treatments have a high recurrence rate and side effects, emphasizing the need for alternative instillations. Previously, we showed that expanded allogeneic human natural killer (NK) cells from peripheral blood are a promising cellular therapy for prostate cancer. However, whether NK cells exhibit a similar killing effect in bladder cancer (BCa) remains unknown. Our objective was to investigate the feasibility and potency of intravesical instillation of expanded allogeneic human NK cells from peripheral blood to treat NMIBC after TURBT. METHODS: Expansion, activation, and cryopreservation of allogeneic human NK cells obtained from peripheral blood were performed as previously described. In vitro cytotoxicity was evaluated using the cell counting kit-8. The levels of perforin, granzyme B, interferon-γ, tumor necrosis factor-α, and chemokines (including C-C-motif ligand (CCL)1, CCL2, CCL20, CCL3L1, CCL4, C-X-C-motif ligand (CXCL)1, CXCL16, CXCL2, CXCL3, CXCL8, and X-motif ligand 1 and 2) were determined using enzyme-linked immunosorbent assay. The expression of CD107a, major histocompatibility complex class I (MHC-I), MHC-I polypeptide-related sequences A and B (MICA/B), cytomegalovirus UL16-binding protein-2/5/6 (ULBP-2/5/6), B7-H6, CD56, CD69, CD25, killer cell Ig-like receptors (KIR)2DL1, KIRD3DL1, NKG2D, NKp30, NKp46, and CD16 of NK cells or BCa and normal urothelial cells were detected using flow cytometry. Cytotoxicity was evaluated using lactate dehydrogenase assay in patient-derived organoid models. BCa growth was monitored in vivo using calipers in male NOD-scid IL2rg−/− mice subcutaneously injected with 5637 with NK cells. Differential gene expressions was investigated using RNA sequence analysis. The chemotaxis of T cells was evaluated using transwell migration assays. RESULTS: We showed that the NK cells possessed higher cytotoxicity against BCa lines with more production of cytokines than normal urothelial cells counterparts in vitro, demonstrated by upregulation of degranulation marker CD107a and increased interferon-γ secretion, by MICA/B/NKG2D and B7H6/NKp30-mediated activation. Furthermore, NK cells demonstrated antitumor effects against BCa in patient-derived organoids and BCa xenograft mouse models. NK cells secreted chemokines, including CCL1/2/20, to induce T-cell chemotaxis when encountering BCa cells. CONCLUSIONS: The expanded NK cells exhibit potent cytotoxicity against BCa cells, with few toxic side effects on normal urothelial cells. In addition, NK cells recruit T cells by secreting a panel of chemokines, which supports the translational application of NK cell intravesical instillation after TURBT from bench to bedside for NMIBC treatment. Source of Funding: This work was supported by grants from National Key R&D Program of China (Grant No. 2022YFE0200800) awarded to Nianzeng Xing, Research Fund for Tumors of Urinary System treatment in China (Grant No. 020) awarded to Fangming Wang © 2024 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 211Issue 5SMay 2024Page: e357 Advertisement Copyright & Permissions© 2024 by American Urological Association Education and Research, Inc.Metrics Author Information Fangming Wang More articles by this author Expand All Advertisement PDF downloadLoading ...