---
title: "GPC3 CAR‑NK Cells Plus Enoblituzumab Improve Anti‑tumor Activity in Hepatocellular Carcinoma"
id: "pubmed-42378857"
canonical_url: "https://medichelpline.com/clinical-feed/pubmed-42378857"
content_type: "clinical_feed_article"
specialty: "Pharmacology"
source_name: "PubMed / NCBI"
source_url: "https://pubmed.ncbi.nlm.nih.gov/42378857/"
doi: "10.1016/j.bbrc.2026.154190"
published_at: "2026-09-03T00:00:00.000Z"
evidence_level: "Journal Article"
license: "CC-BY-NC-4.0 / Informational Use"
---
# GPC3 CAR‑NK Cells Plus Enoblituzumab Improve Anti‑tumor Activity in Hepatocellular Carcinoma
## Provenance & Clinical Metadata
- **Canonical URL:** https://medichelpline.com/clinical-feed/pubmed-42378857
- **Specialty:** [Pharmacology](https://medichelpline.com/clinical-feed/pharmacology.md)
- **Primary Source:** PubMed / NCBI
- **Source URL:** [Original Journal Publication](https://pubmed.ncbi.nlm.nih.gov/42378857/)
- **DOI:** [10.1016/j.bbrc.2026.154190](https://doi.org/10.1016%2Fj.bbrc.2026.154190)
- **Published At:** 2026-09-03T00:00:00.000Z
- **Evidence Rating:** Journal Article
## Executive GIST (TL;DR)
- Hepatocellular carcinoma (HCC) causes high mortality and presents challenges for cell therapies due to antigen escape in solid tumors. - The study evaluated a dual‑targeting immunotherapy combining **GPC3** chimeric antigen receptor **(CAR)-NK** cells with the B7H3‑blocking monoclonal antibody **Enoblituzumab**. - Bioinformatic database analysis and flow cytometry confirmed co‑expression of **GPC3** and **B7H3** in HCC tissues and cell lines. - Human PBMC‑derived NK cells were expanded and engineered to express a **GPC3 CAR** for functional studies. - In vitro cytotoxicity was measured by LDH release and real‑time cell analysis; NK activation assessed by **IFN-γ** secretion (ELISA) and **CD107a** degranulation (flow cytometry). - **GPC3 CAR‑NK** cells showed potent cytotoxicity against HCC cells in vitro; **Enoblituzumab** produced strong killing against **B7H3**‑positive tumors. - The combination of **GPC3 CAR‑NK** cells plus **Enoblituzumab** produced synergistic increases in cytotoxicity, **IFN‑γ** production, and **CD107a** degranulation compared with either agent alone. - In vivo efficacy was tested in NCG mice bearing subcutaneous Huh7 tumors; combination therapy significantly suppressed tumor growth without causing weight loss or splenomegaly, indicating favorable safety in this model. - High co‑expression of **GPC3** and **B7H3** correlated with poorer patient prognosis in database analysis. - The authors conclude that dual targeting of **GPC3** and **B7H3** augments NK cell–mediated antitumor activity and supports development of dual‑target immunotherapies for HCC. - Details on dosing, sample sizes, long‑term toxicity, and clinical translation were not reported in the abstract.
## Clinical Analysis & Structured Key Points
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Epub 2026 Jun 25. # GPC3 chimeric antigen receptor (CAR)-NK cells combined with Enoblituzumab enhance the anti-tumor efficacy against hepatocellular carcinoma [Wen Zhuang](https://pubmed.ncbi.nlm.nih.gov/?term=Zhuang+W&cauthor_id=42378857)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42378857/#full-view-affiliation-1 "Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China."), [Guanyu Xiao](https://pubmed.ncbi.nlm.nih.gov/?term=Xiao+G&cauthor_id=42378857)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42378857/#full-view-affiliation-1 "Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China."), [Mengyao Zhang](https://pubmed.ncbi.nlm.nih.gov/?term=Zhang+M&cauthor_id=42378857)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42378857/#full-view-affiliation-1 "Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China."), [Jie Wang](https://pubmed.ncbi.nlm.nih.gov/?term=Wang+J&cauthor_id=42378857)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42378857/#full-view-affiliation-1 "Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China."), [Wenjing Tang](https://pubmed.ncbi.nlm.nih.gov/?term=Tang+W&cauthor_id=42378857)[ 2 ](https://pubmed.ncbi.nlm.nih.gov/42378857/#full-view-affiliation-2 "Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China. Electronic address: tangwj0314@126.com."), [Yong Liang](https://pubmed.ncbi.nlm.nih.gov/?term=Liang+Y&cauthor_id=42378857)[ 3 ](https://pubmed.ncbi.nlm.nih.gov/42378857/#full-view-affiliation-3 "Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China. Electronic address: Yongl@xzhmu.edu.cn.") Affiliations Expand ### Affiliations * 1 Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China. * 2 Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China. Electronic address: tangwj0314@126.com. * 3 Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China. Electronic address: Yongl@xzhmu.edu.cn. * PMID: **42378857** * DOI: [ 10.1016/j.bbrc.2026.154190 ](https://doi.org/10.1016/j.bbrc.2026.154190) Item in Clipboard # GPC3 chimeric antigen receptor (CAR)-NK cells combined with Enoblituzumab enhance the anti-tumor efficacy against hepatocellular carcinoma Wen Zhuang et al. Biochem Biophys Res Commun. 2026. Show details Display options Display options Format Abstract PubMed PMID Biochem Biophys Res Commun Actions * [ Search in PubMed ](https://pubmed.ncbi.nlm.nih.gov/?term=%22Biochem+Biophys+Res+Commun%22%5Bjour%5D&sort=date&sort_order=desc) * [ Search in NLM Catalog ](https://www.ncbi.nlm.nih.gov/nlmcatalog?term=%22Biochem+Biophys+Res+Commun%22%5BTitle+Abbreviation%5D) * [ Add to Search ](https://pubmed.ncbi.nlm.nih.gov/42378857/) . 2026 Sep 3:829:154190. doi: 10.1016/j.bbrc.2026.154190. Epub 2026 Jun 25. ### Authors [Wen Zhuang](https://pubmed.ncbi.nlm.nih.gov/?term=Zhuang+W&cauthor_id=42378857)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42378857/#short-view-affiliation-1 "Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China."), [Guanyu Xiao](https://pubmed.ncbi.nlm.nih.gov/?term=Xiao+G&cauthor_id=42378857)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42378857/#short-view-affiliation-1 "Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China."), [Mengyao Zhang](https://pubmed.ncbi.nlm.nih.gov/?term=Zhang+M&cauthor_id=42378857)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42378857/#short-view-affiliation-1 "Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China."), [Jie Wang](https://pubmed.ncbi.nlm.nih.gov/?term=Wang+J&cauthor_id=42378857)[ 1 ](https://pubmed.ncbi.nlm.nih.gov/42378857/#short-view-affiliation-1 "Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China."), [Wenjing Tang](https://pubmed.ncbi.nlm.nih.gov/?term=Tang+W&cauthor_id=42378857)[ 2 ](https://pubmed.ncbi.nlm.nih.gov/42378857/#short-view-affiliation-2 "Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China. Electronic address: tangwj0314@126.com."), [Yong Liang](https://pubmed.ncbi.nlm.nih.gov/?term=Liang+Y&cauthor_id=42378857)[ 3 ](https://pubmed.ncbi.nlm.nih.gov/42378857/#short-view-affiliation-3 "Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China. Electronic address: Yongl@xzhmu.edu.cn.") ### Affiliations * 1 Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China. * 2 Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China. Electronic address: tangwj0314@126.com. * 3 Huaian second people's hospital/The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, Jiangsu, 223002, China. Electronic address: Yongl@xzhmu.edu.cn. * PMID: **42378857** * DOI: [ 10.1016/j.bbrc.2026.154190 ](https://doi.org/10.1016/j.bbrc.2026.154190) Item in Clipboard Full text links Cite Display options Display options Format Abstract PubMed PMID ## Abstract Hepatocellular carcinoma (HCC) is a leading cause of cancer mortality worldwide. Although CAR-NK cell therapies have shown promise in hematologic malignancies, their efficacy in solid tumors like HCC is often limited by antigen escape. Here, we investigated a dual-targeting strategy combining GPC3 CAR-NK cells with the B7H3-blocking monoclonal antibody Enoblituzumab. We confirmed GPC3 and B7H3 co-expression in HCC tissues and cell lines via database analysis and flow cytometry. GPC3 CAR-NK cells were generated from expanded human PBMC-derived NK cells. In vitro cytotoxicity against HCC cells was assessed by LDH release and real-time cell analysis, while IFN-γ secretion and CD107a degranulation were measured by ELISA and flow cytometry. In vivo antitumor efficacy was evaluated in NCG mice bearing subcutaneous Huh7 tumors. High GPC3 and B7H3 co-expression correlated with poor patient prognosis. GPC3 CAR-NK cells exhibited potent cytotoxicity against HCC cells in vitro. Enoblituzumab triggered strong killing effects against B7H3-positive tumors. Notably, the combination synergistically enhanced cytotoxicity, IFN-γ production, and CD107a degranulation compared to either monotherapy. In vivo, combination treatment significantly suppressed tumor growth without inducing weight loss or splenomegaly, demonstrating favorable efficacy and safety. Our findings demonstrate that dual targeting of GPC3 and B7H3 effectively enhances NK cell-mediated antitumor activity. This strategy leverages both CAR-mediated specificity and NK cell-intrinsic mechanisms, providing a strong rationale for developing dual-target immunotherapies to improve outcomes in HCC patients. **Keywords:** B7H3; CAR-NK cells; Dual targets; GPC3; Hepatocellular carcinoma; Synergistic effects. Copyright © 2026 Elsevier Inc. All rights reserved. [PubMed Disclaimer](https://pubmed.ncbi.nlm.nih.gov/disclaimer/) ## Conflict of interest statement Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Yong Liang reports financial support was provided by National Natural Science Foundation of China. Yong Liang reports a relationship with Huaian Second People's Hospital that includes: employment. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. ## Similar articles * [ Exploring Glypican-3 targeted CAR-NK treatment and potential therapy resistance in hepatocellular carcinoma. ](https://pubmed.ncbi.nlm.nih.gov/39841705/) Yang L, Pham K, Xi Y, Wu Q, Liu D, Robertson KD, Liu C.Yang L, et al.PLoS One. 2025 Jan 22;20(1):e0317401. doi: 10.1371/journal.pone.0317401. eCollection 2025.PLoS One. 2025.PMID: 39841705Free PMC article. * [ Development of glypican-3-specific chimeric antigen receptor-modified natural killer cells and optimization as a therapy for hepatocellular carcinoma. ](https://pubmed.ncbi.nlm.nih.gov/38922297/) Cao B, Ni Q, Chen Z, Yang S, Zhang X, Su H, Zhang Z, Zhao Q, Zhu X, Liu M.Cao B, et al.J Leukoc Biol. 2025 Feb 13;117(2):qiae144. doi: 10.1093/jleuko/qiae144.J Leukoc Biol. 2025.PMID: 38922297 * [ Chimeric antigen receptor-T cells targeting AFP-GPC3 mediate increased antitumor efficacy in hepatocellular carcinoma. ](https://pubmed.ncbi.nlm.nih.gov/39757079/) Li M, Chen T, Huang R, Cen Y, Zhao F, Fan R, He G.Li M, et al.Arab J Gastroenterol. 2025 Feb;26(1):84-93. doi: 10.1016/j.ajg.2024.12.002. Epub 2025 Jan 4.Arab J Gastroenterol. 2025.PMID: 39757079 * [ Glypican-3-Specific CAR NK Cells Co-Secreting IL-15 and IFN-α Have Increased Anti-Tumor Function Versus Hepatocellular Carcinoma In Vitro. ](https://pubmed.ncbi.nlm.nih.gov/41465318/) Busà R, Iannolo G, Douradinha B, Pagano D, Gallina A, Cappello G, La Rocca A, Gruttadauria S, Conaldi PG, Badami E.Busà R, et al.Int J Mol Sci. 2025 Dec 10;26(24):11892. doi: 10.3390/ijms262411892.Int J Mol Sci. 2025.PMID: 41465318Free PMC article. * [ A Review of Recent Advances in Chimeric Antigen Receptor (CAR) T-Cell Therapy for Hepatocellular Carcinoma. ](https://pubmed.ncbi.nlm.nih.gov/42332855/) Lv X, Chen X, Ge Y, Si G, Li Y, Li X, Yuan X.Lv X, et al.Med Sci Monit. 2026 Jun 23;32:e953528. doi: 10.12659/MSM.953528.Med Sci Monit. 2026.PMID: 42332855Free PMC article.Review. 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