Construction of humanized anti-HER2 single-chain variable fragments (husFvs) and achievement of potent tumor suppression with the reconstituted husFv-Fdt-tBid immunoapoptotin

Ouyang, Qing; Yan, Bo; Li, Ang; Hu, Zhi-Song; Feng, Jie; Lun, Xin-Xin; Zhang, Mingming; Zhang, Mengde; Wu, Kaichun; Xue, Fangzheng; Yang, Angang; Zhao, Jing

doi:10.1016/j.biomaterials.2018.06.016

Cited by 11 publications

(13 citation statements)

References 32 publications

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“…The docking analysis also revealed that all the scFvs formed distinct but overlapping interfaces with domain IV of the HER2 ECD, which indicated that the manipulation of the FRs mildly influenced the binding specificity of the scFvs. A potent application of scFvs in tumour diagnosis and treatment requires the administration of scFvs at a dose sufficient for tumour targeting with minimal off-tumour effects (5). The improved affinity of EX1 for efficient HER2 binding required a small dose, which indicated that it poses a reduced risk of off-target effects and is less likely to induce toxicity in organs, such as the liver and kidney.…”

Section: Discussionmentioning

confidence: 99%

“…e23sFv has been indicated to exhibit reduced molecular weight and more efficient penetration into solid tumours compared with that of e23; therefore, it has been utilized in immunoconjugates, such as immunotoxin or immune-proapoptotic molecules, as the portion targeted to HER2 (3,4). Despite its advantages in terms of size and trafficking, e23sFv exhibits a 4-fold reduced binding affinity for HER2 compared with that of e23 or the corresponding antigen-binding fragment (Fab) (5). To achieve equivalent effectiveness, a higher dose is required for antibodies with less affinity; however, an increased dose of therapeutic antibodies can result in more intensive immunogenic reactions and possibly unexpected toxicities (6).…”

Section: Introductionmentioning

confidence: 99%

“…To achieve equivalent effectiveness, a higher dose is required for antibodies with less affinity; however, an increased dose of therapeutic antibodies can result in more intensive immunogenic reactions and possibly unexpected toxicities (6). To ensure the safety and efficacy of e23sFv-derived immuno-proapoptotic molecules in HER2-targeted tumour treatment, it is necessary to enhance the affinity of e23sFv for the HER2 extracellular domain (ECD) (5).…”

Section: Introductionmentioning

confidence: 99%

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Syngeneic homograft of framework regions enhances the affinity of the mouse anti‑human epidermal receptor 2 single‑chain antibody e23sFv

et al. 2020

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e23sFv is a HER2-targeted single-chain variable fragment (scFV) that was characterized as the targeting portion of a HER2-targeted tumour proapoptotic molecule in our previous study. In vitro antibody affinity maturation is a method to enhance antibody affinity either by complementarity-determining region (CDR) mutagenesis or by framework region (FR) engraftment. In the present study, the affinity of e23sFv was enhanced using two strategies. In one approach, site-directed mutations were introduced into the FRs of e23sFv (designated EMEY), and in the other approach e23sFv FRs were substituted with FRs from the most homologous screened antibodies (designated EX1 and EX2). Notably, EX1 derived from the FR engraftment strategy demonstrated a 4-fold higher affinity for HER2 compared with e23sFv and was internalized into HER2-overexpressing cells; however, EMEY and EX2 exhibited reduced affinity for HER2 and decreased internalization potential compared with EX1. The 3D structure of EX1 and the HER2-EX1 complex was acquired using molecular homology modelling and docking and the HER2 epitopes of EX1 and the molecular interaction energy of the EX1-HER2 complex were predicted. In the present study, it was demonstrated that scFv affinity improvement based on sequence alignment was feasible and effective. Moreover, the FR grafting strategy was indicated to be more effective and simple compared with site-directed mutagenesis to improve e23sFv affinity. In conclusion, it was indicated that the affinity-improved candidate EX1 may present a great potential for the diagnosis and treatment of HER2-overexpressing tumours.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Syngeneic homograft of framework regions enhances the affinity of the mouse anti‑human epidermal receptor 2 single‑chain antibody e23sFv

et al. 2020

Self Cite

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“…Another type of complete humanization of rITs is named human cytolytic fusion proteins (hCFPs), originating from the cytotoxic endogenous human apoptotic-relevant enzymes fused to humanized antibody fragments. So far, various endogenous pro-apoptotic molecules such as granzyme B (GrB) [ 88 ], truncated Bid (tBid) [ 94 ], caspase-3 [ 93 ], caspase-6 [ 92 , 146 ] and apoptosis-inducing factor (AIF) [ 147 ] have been utilized to induce apoptosis in tumor cells (Table 2 ).…”

Section: Her2-specific Immunotoxinsmentioning

confidence: 99%

“…Humanization of e23sFv (husFvs) increased its affinity to recombinant HER2 up to 94-fold. In addition, the immunogenicity of e23sFv was considerably reduced after humanization [ 94 ].…”

Section: Her2-specific Immunotoxinsmentioning

confidence: 99%

Recombinant immunotoxins development for HER2-based targeted cancer therapies

et al. 2021

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Understanding the molecular mechanisms of cancer biology introduces targeted therapy as a complementary method along with other conventional therapies. Recombinant immunotoxins are tumor specific antibodies that their recognizing fragment is utilized for delivering modified toxins into tumor cells. These molecules have been considered as a targeted strategy in the treatment of human cancers. HER2 tumor biomarker is a transmembrane tyrosine kinase receptor that can be used for targeted therapies in the forms of anti-HER2 monoclonal antibodies, antibody–drug conjugates and immunotoxins. There have been many studies on HER2-based immunotoxins in recent years, however, little progress has been made in the clinical field which demanded more improvements. Here, we summarized the HER2 signaling and it’s targeting using immunotherapeutic agents in human cancers. Then, we specifically reviewed anti-HER2 immunotoxins, and their strengths and drawbacks to highlight their promising clinical impact.

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Targeting BRIX1 via Engineered Exosomes Induces Nucleolar Stress to Suppress Cancer Progression

Gan,

Hao,

Han

et al. 2024

Advanced Science

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Elevated ribosome biogenesis correlates with the rapid growth and progression of cancer. Targeted blockade of ribosome biogenesis induces nucleolar stress, which preferentially leads to the elimination of malignant cells. In this study, it is reported that the nucleolar protein BRIX1 is a critical regulator for the homeostasis between ribosome biogenesis and p53 activation. BRIX1 facilitated the processing of pre‐rRNA by supporting the formation of the PeBoW complex. In addition, BRIX1 prevented p53 activation in response to nucleolar stress by impairing the interactions between MDM2 and the ribosomal proteins, RPL5, and RPL11, thereby triggering the resistance of cancer cells to chemotherapy. Conversely, depletion of BRIX1 induced nucleolar stress, which in turn activated p53 through RPL5 and RPL11, consequently inhibiting the growth of tumors. Moreover, engineered exosomes are developed, which are surface‐decorated with iRGD, a tumor‐homing peptide, and loaded with siRNAs specific to BRIX1, for the treatment of cancer. iRGD‐Exo‐siBRIX1 significantly suppressed the growth of colorectal cancer and enhanced the efficacy of 5‐FU chemotherapy in vivo. Overall, the study uncovers that BRIX1 functions as an oncoprotein to promote rRNA synthesis and dampen p53 activity, and also implies that targeted inhibition of BRIX1 via engineered exosomes can be a potent approach for cancer therapy.

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Construction of humanized anti-HER2 single-chain variable fragments (husFvs) and achievement of potent tumor suppression with the reconstituted husFv-Fdt-tBid immunoapoptotin

Cited by 11 publications

References 32 publications

Syngeneic homograft of framework regions enhances the affinity of the mouse anti‑human epidermal receptor 2 single‑chain antibody e23sFv

Syngeneic homograft of framework regions enhances the affinity of the mouse anti‑human epidermal receptor 2 single‑chain antibody e23sFv

Recombinant immunotoxins development for HER2-based targeted cancer therapies

Targeting BRIX1 via Engineered Exosomes Induces Nucleolar Stress to Suppress Cancer Progression

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