A Novel Vaccine Targeting Glypican-3 as a Treatment for Hepatocellular Carcinoma

Wu, Qiang-Sheng; Pi, Liya; Trinh, Thu Le; Zuo, Chaohui; Xia, Man; Jiao, Yu; Hou, Zhouhua; Jo, Sung Duk; Puszyk, William; Pham, Kien; Nelson, David R.; Robertson, Keith D.; Ostrov, David A.; Rameshwar, Pranela; Xia, Chang; Liu, Chen

doi:10.1016/j.ymthe.2017.08.005

Cited by 26 publications

(23 citation statements)

References 56 publications

(77 reference statements)

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“…In mice the opposite is found, with the overexpression of GPC3 in the liver resulting in lower liver cell proliferation and a lack of correlation with HCC formation . The mouse HCC cell line Hepa1‐6 has been reported to express GPC3, but we found that this cell line was not sensitive to immunotoxin treatment (Supporting Fig. ).…”

Section: Discussionmentioning

confidence: 66%

Engineered Anti‐GPC3 Immunotoxin, HN3‐ABD‐T20, Produces Regression in Mouse Liver Cancer Xenografts Through Prolonged Serum Retention

et al. 2020

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Background and Aims Treatment of hepatocellular carcinomas using our glypican‐3 (GPC3)‐targeting human nanobody (HN3) immunotoxins causes potent tumor regression by blocking protein synthesis and down‐regulating the Wnt signaling pathway. However, immunogenicity and a short serum half‐life may limit the ability of immunotoxins to transition to the clinic. Approach and Results To address these concerns, we engineered HN3‐based immunotoxins to contain various deimmunized Pseudomonas exotoxin (PE) domains. This included HN3‐T20, which was modified to remove T‐cell epitopes and contains a PE domain II truncation. We compared them to our previously reported B‐cell deimmunized immunotoxin (HN3‐mPE24) and our original HN3‐immunotoxin with a wild‐type PE domain (HN3‐PE38). All of our immunotoxins displayed high affinity to human GPC3, with HN3‐T20 having a KD value of 7.4 nM. HN3‐T20 retained 73% enzymatic activity when compared with the wild‐type immunotoxin in an adenosine diphosphate–ribosylation assay. Interestingly, a real‐time cell growth inhibition assay demonstrated that a single dose of HN3‐T20 at 62.5 ng/mL (1.6 nM) was capable of inhibiting nearly all cell proliferation during the 10‐day experiment. To enhance HN3‐T20’s serum retention, we tested the effect of adding a streptococcal albumin‐binding domain (ABD) and a llama single‐domain antibody fragment specific for mouse and human serum albumin. For the detection of immunotoxin in mouse serum, we developed a highly sensitive enzyme‐linked immunosorbent assay and found that HN3‐ABD‐T20 had a 45‐fold higher serum half‐life than HN3‐T20 (326 minutes vs. 7.3 minutes); consequently, addition of an ABD resulted in HN3‐ABD‐T20–mediated tumor regression at 1 mg/kg. Conclusion These data indicate that ABD‐containing deimmunized HN3‐T20 immunotoxins are high‐potency therapeutics ready to be evaluated in clinical trials for the treatment of liver cancer.

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Section: Discussionmentioning

confidence: 66%

Engineered Anti‐GPC3 Immunotoxin, HN3‐ABD‐T20, Produces Regression in Mouse Liver Cancer Xenografts Through Prolonged Serum Retention

et al. 2020

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“…19 Antibodies against GPC3 and GPC3-peptide vaccines have been demonstrated to be safe and effective. 36,37 Presently, CAR-NK-92 against GPC3 especially killed GPC3 + HCC cells and did not harm GPC3 − normal hepatocellular cells, suggesting that GPC3 is a suitable target in CAR-NK therapy.…”

Section: Discussionmentioning

confidence: 93%

<p>DNAM1 and 2B4 Costimulatory Domains Enhance the Cytotoxicity of Anti-GPC3 Chimeric Antigen Receptor-Modified Natural Killer Cells Against Hepatocellular Cancer Cells in vitro</p>

Huang

Zeng

et al. 2020

CMAR

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Purpose: Hepatocellular cancer (HCC) is the sixth most prevalent cancer and the third leading cause of cancer-related death worldwide. Cellular immunotherapy against glypican 3 (GPC3) has recently been used in the treatment of HCC, following the success of chimeric antigen receptor (CAR)-T therapy in treatment of B cell malignancy. However, CART cells are not "off-the-shelf" and always cause cytokine release syndrome, which can be eliminated by using natural killer (NK) cells as effector cells. Since a costimulatory signal is necessary for the activation, persistence, or cytotoxicity of CART cells, we speculated that the costimulatory signal is also required for CAR-NK cells in HCC treatment. Methods: Five anti-GPC3 CAR plasmids containing different costimulatory domains were constructed. They included Z (only the CD3ζ domain, no costimulatory domain), CD28.Z (T-cell costimulatory domain CD28), DNAM1/2B4.Z (NK-cell-associated costimulatory domain DNAM1 or 2B4), and DNAM1.2B4.Z (both NK-cell-associated costimulatory domains). Respective CAR-NK-92 cells were generated. The MTT viability assay was performed to evaluate the effect of the different costimulatory domains on CAR-NK-cell proliferation. The effect on persistence was analyzed using an apoptosis assay and flow cytometry. Special cytotoxicity against normal hepatocellular cells and GPC3 + malignant cells was investigated in vitro. The concentration of cytokines (TNF-α and IFN-γ) released by CAR-NK-92 cells was also measured by ELISA. Results: NK-cell-associated costimulatory signal was necessary for CAR-NK-92 cells. CAR-NK-92 cells with DNAM1 and/or 2B4 expanded more quickly and persisted with a lower apoptotic ratio, compared to the presence of CD28 or no costimulatory signal. All CAR-NK-92 cells showed special cellular cytotoxicity in vitro. CAR-NK-92 cells with NK-cell-associated costimulatory domains exhibited higher cytotoxic ability compared with those without any costimulatory domain or with T-cell costimulatory domain. CAR-NK-92 cells with both DNAM1 and 2B4 displayed the highest cytotoxicity. The cytokine release assay results were consistent with those of the cytotoxicity assay. Conclusion: We provided the first evidence supporting a strategy using DNAM1 and 2B4 costimulatory domains to generate anti-GPC3 CAR-NK-92 cells, which exhibits enhanced cytotoxicity against hepatocellular cancer cells in vitro.

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“…In addition to confirming these established signaling interactions, our in silico screen suggests putative interactions which may provide novel therapeutic targets for GBM. Binding of GPC3 to IGF1R or CD81 is known to contribute to hepatocellular carcinoma development and invasiveness 41 ; immunotherapies targeting GPC3, which are showing promise in hepatocellular carcinoma, 42 , 43 may also benefit GBM patients. Our results also suggest AFDN/MLLT4 (adherens junction formation factor/myeloid/lymphoid or mixed-lineage leukemia) as a ligand activating ephrin receptor A7, which has been linked to adverse outcome in primary and recurrent GBM.…”

Section: Discussionmentioning

confidence: 99%

Modeling glioblastoma invasion using human brain organoids and single-cell transcriptomics

et al. 2020

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Background Glioblastoma (GBM) consists of devastating neoplasms with high invasive capacity, which have been difficult to study in vitro in a human-derived model system. Therapeutic progress is also limited by cellular heterogeneity within and between tumors, among other factors such as therapy resistance. To address these challenges, we present an experimental model using human cerebral organoids as a scaffold for patient-derived GBM cell invasion. Methods This study combined tissue clearing and confocal microscopy with single-cell RNA sequencing of GBM cells before and after co-culture with organoid cells. Results We show that tumor cells within organoids extend a network of long microtubes, recapitulating the in vivo behavior of GBM. Transcriptional changes implicated in the invasion process are coherent across patient samples, indicating that GBM cells reactively upregulate genes required for their dispersion. Potential interactions between GBM and organoid cells identified by an in silico receptor–ligand pairing screen suggest functional therapeutic targets. Conclusions Taken together, our model has proven useful for studying GBM invasion and transcriptional heterogeneity in vitro, with applications for both pharmacological screens and patient-specific treatment selection on a time scale amenable to clinical practice. Key Points 1. Organoid technology and single-cell transcriptomics reveal cellular interactions of invasive GBM cells. 2. Transcriptional changes implicated during invasion suggest novel therapeutic targets for GBM. 3. Time scales are amenable to clinical practice and high-content drug screens.

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A Novel Vaccine Targeting Glypican-3 as a Treatment for Hepatocellular Carcinoma

Cited by 26 publications

References 56 publications

Engineered Anti‐GPC3 Immunotoxin, HN3‐ABD‐T20, Produces Regression in Mouse Liver Cancer Xenografts Through Prolonged Serum Retention

Engineered Anti‐GPC3 Immunotoxin, HN3‐ABD‐T20, Produces Regression in Mouse Liver Cancer Xenografts Through Prolonged Serum Retention

<p>DNAM1 and 2B4 Costimulatory Domains Enhance the Cytotoxicity of Anti-GPC3 Chimeric Antigen Receptor-Modified Natural Killer Cells Against Hepatocellular Cancer Cells in vitro</p>

Modeling glioblastoma invasion using human brain organoids and single-cell transcriptomics

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