Human Blood Serum Inhibits Ductal Carcinoma Cells BT474 Growth and Modulates Effect of HER2 Inhibition

Kamashev, Dmitrii; Shaban, Nina; Suntsova, Maria; Raevskiy, Mikhail; Efimov, Victor; Moisseev, Aleksey; Buzdin, Anton

doi:10.3390/biomedicines10081914

Cited by 8 publications

(7 citation statements)

References 78 publications

(97 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The difference between SKBR3 and BT474 responses may be explained by the differential total expression of HER2, as well as the expression of EGFR and HER3 relative to HER2. BT474 expresses more HER2 ( ~ 2.8 × 10 6 copies per cell) 53 relative to SKBR3 ( ~ 1.2 × 10 6 copies) 54 . Both express HER3 to a similar level (1-1.2 × 10 6 copies) but SKBR3 expresses ~3-fold more EGFR (2.2 × 10 5 copies) than BT474 (7 × 10 4 copies) 55 .…”

Section: Discussionmentioning

confidence: 99%

SpyMask enables combinatorial assembly of bispecific binders

Driscoll,

Keeble,

Howarth

2024

Nat Commun

View full text Add to dashboard Cite

Bispecific antibodies are a successful and expanding therapeutic class. Standard approaches to generate bispecifics are complicated by the need for disulfide reduction/oxidation or specialized formats. Here we present SpyMask, a modular approach to bispecifics using SpyTag/SpyCatcher spontaneous amidation. Two SpyTag-fused antigen-binding modules can be precisely conjugated onto DoubleCatcher, a tandem SpyCatcher where the second SpyCatcher is protease-activatable. We engineer a panel of structurally-distinct DoubleCatchers, from which binders project in different directions. We establish a generalized methodology for one-pot assembly and purification of bispecifics in 96-well plates. A panel of binders recognizing different HER2 epitopes were coupled to DoubleCatcher, revealing unexpected combinations with anti-proliferative or pro-proliferative activity on HER2-addicted cancer cells. Bispecific activity depended sensitively on both binder orientation and DoubleCatcher scaffold geometry. These findings support the need for straightforward assembly in different formats. SpyMask provides a scalable tool to discover synergy in bispecific activity, through modulating receptor organization and geometry.

show abstract

Section: Discussionmentioning

confidence: 99%

SpyMask enables combinatorial assembly of bispecific binders

Driscoll,

Keeble,

Howarth

2024

Nat Commun

View full text Add to dashboard Cite

show abstract

“…This result may be explained by the differential total expression of HER2, as well as the expression of Epidermal Growth Factor Receptor (EGFR) and HER3 relative to HER2. BT474 expresses HER2 to a greater extent (∼2.8 × 10 6 copies per cell) (Kamashev et al, 2022) relative to SKBR3 (∼1.2 × 10 6 copies) (Hendriks et al, 2013). Both express HER3 to a similar level (1-1.2 × 10 6 copies) but SKBR3 expresses ∼3-fold more EGFR (2.2 × 10 5 copies) than BT474 (7 × 10 4 copies) (Brockhoff et al, 2001).…”

Section: Discussionmentioning

confidence: 99%

SpyMask Enables Combinatorial Assembly of Bispecific Binders

Driscoll,

Keeble,

Howarth

2023

Preprint

View full text Add to dashboard Cite

Bispecific antibodies are a successful and expanding therapeutic class, bridging two cell-types or engaging two different molecules on the same cell. Bispecifics unlock avenues towards synergy, resistance evasion, and signaling bias. Standard approaches to generate bispecifics are complicated by the need for disulfide reduction/oxidation or cloning of each binder molecule in two different formats. Here we present a modular approach to bispecifics using SpyTag/SpyCatcher spontaneous amidation, where all binders are cloned in the same format, bearing a SpyTag. Two SpyTag-fused antigen-binding modules can be precisely conjugated onto DoubleCatcher, a tandem SpyCatcher where the second Catcher is unreactive until unveiling of reactivity using site-specific protease. Assembly on DoubleCatcher is efficient in phosphate-buffered saline at 37 °C, with half-times less than 5 min for both SpyCatcher arms and over 97% bispecific homogeneity. We engineer a panel of DoubleCatchers, locked through disulfide bonds to direct binders to project in different directions from the hub. We establish a generalized methodology for one-pot assembly and purification of bispecifics in 96-well plate format. A panel of Fab, affibody or nanobody binders recognizing different sites on HER2 were coupled to DoubleCatcher, revealing unexpected combinations with anti-proliferative or pro-proliferative activity on HER2-addicted cancer cells. Bispecific activity depended sensitively on both the order of the binders within the assembly and the geometry of DoubleCatcher scaffolds. These findings support the need for straightforward assembly in different formats. SpyCombinator provides a simple and scalable tool to discover synergy in bispecific activity, through modulating receptor organization and geometry.

show abstract

“…The analysis of molecular pathways revealed that the addition of human serum reactivated MAPK signaling pathways inhibited by cetuximab alone [ 90 ]. Interestingly, human serum also modulated the effect of the HER2 monoclonal antibody trastuzumab on the HER2-overexpressing cell line BT-474 [ 223 ].…”

Section: Egfr-specific Therapeutic Monoclonal Antibodiesmentioning

confidence: 99%

Targeted Inhibitors of EGFR: Structure, Biology, Biomarkers, and Clinical Applications

Shaban,

Kamashev,

Emelianova

et al. 2023

Cells

Self Cite

View full text Add to dashboard Cite

Members of the EGFR family of tyrosine kinase receptors are major regulators of cellular proliferation, differentiation, and survival. In humans, abnormal activation of EGFR is associated with the development and progression of many cancer types, which makes it an attractive target for molecular-guided therapy. Two classes of EGFR-targeted cancer therapeutics include monoclonal antibodies (mAbs), which bind to the extracellular domain of EGFR, and tyrosine kinase inhibitors (TKIs), which mostly target the intracellular part of EGFR and inhibit its activity in molecular signaling. While EGFR-specific mAbs and three generations of TKIs have demonstrated clinical efficacy in various settings, molecular evolution of tumors leads to apparent and sometimes inevitable resistance to current therapeutics, which highlights the need for deeper research in this field. Here, we tried to provide a comprehensive and systematic overview of the rationale, molecular mechanisms, and clinical significance of the current EGFR-targeting drugs, highlighting potential candidate molecules in development. We summarized the underlying mechanisms of resistance and available personalized predictive approaches that may lead to improved efficacy of EGFR-targeted therapies. We also discuss recent developments and the use of specific therapeutic strategies, such as multi-targeting agents and combination therapies, for overcoming cancer resistance to EGFR-specific drugs.

show abstract

Human Blood Serum Inhibits Ductal Carcinoma Cells BT474 Growth and Modulates Effect of HER2 Inhibition

Cited by 8 publications

References 78 publications

SpyMask enables combinatorial assembly of bispecific binders

SpyMask enables combinatorial assembly of bispecific binders

SpyMask Enables Combinatorial Assembly of Bispecific Binders

Targeted Inhibitors of EGFR: Structure, Biology, Biomarkers, and Clinical Applications

Contact Info

Product

Resources

About