Targeting ITGβ3 to Overcome Trastuzumab Resistance through Epithelial–Mesenchymal Transition Regulation in HER2-Positive Breast Cancer

International Journal of Molecular Sciences, Год журнала: 2024, Номер 25(16), С. 8640 - 8640

Опубликована: Авг. 8, 2024

HER2-positive breast cancer, representing 15–20% of all cancer cases, often develops resistance to the HER2-targeted therapy trastuzumab. Unfortunately, effective treatments for advanced remain scarce. This study aims investigate roles ITGβ3, and Hedgehog signaling in trastuzumab explore potential combining with cilengitide as a therapeutic strategy. Quantitative gene expression analysis was performed assess transcription EMT (epithelial–mesenchymal transition) markers Slug, Snail, Twist2, Zeb1 trastuzumab-resistant cells. The effects ITGβ3 were investigated. Additionally, combination evaluated. Acquired induced Zeb1, indicating increased EMT. mediated by ITGB3 signaling. regulated both pathway EMT, latter being independent pathway. showed synergistic effect, reducing activity. Targeting cilengitide, combined trastuzumab, effectively suppresses offering strategy overcome improve outcomes patients.

Язык: Английский

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HER2-Displaying M13 Bacteriophages induce Therapeutic Immunity against Breast Cancer

Cancers, Год журнала: 2022, Номер 14(16), С. 4054 - 4054

Опубликована: Авг. 22, 2022

The advent of trastuzumab has significantly improved the prognosis HER2-positive (HER2+) breast cancer patients; nevertheless, drug resistance limits its clinical benefit. Anti-HER2 active immunotherapy represents an attractive alternative strategy, but effective immunization needs to overcome patient's immune tolerance against self-HER2. Phage display technology, taking advantage phage intrinsic immunogenicity, permits one generate vaccines able break self-antigens. In this study, we demonstrate that both preventive and therapeutic vaccination with M13 bacteriophages, displaying extracellular (EC) transmembrane (TM) domains human HER2 or Δ16HER2 splice variant on their surface (ECTM Δ16ECTM phages), delayed mammary tumor onset reduced growth rate multiplicity in ∆16HER2 transgenic mice, which are tolerant ∆16HER2. This antitumor protection correlated anti-HER2 antibody production. molecular mechanisms underlying anticancer effect vaccine-elicited antibodies were analyzed vitro BT-474 cells, sensitive resistant trastuzumab. Immunoglobulins (IgG) purified from sera cell viability mainly by impairing ERK phosphorylation reactivating retinoblastoma protein function trastuzumab-sensitive -resistant cells. conclusion, demonstrated phage-based impair progression, opening new perspectives for HER2+ treatment.

Язык: Английский

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Highly Multiplexing, Throughput and Efficient Single‐Cell Protein Analysis with Digital Microfluidics

Small Methods, Год журнала: 2024, Номер unknown

Опубликована: Апрель 12, 2024

Abstract Proteins as crucial components of cells are responsible for the majority cellular processes. Sensitive and efficient protein detection enables a more accurate comprehensive investigation phenotypes life activities. Here, sequencing method with high multi p lexing, th ro ughpu t , c e ll utilization, in tegration based on digital microfluidics (DMF‐Protein‐seq) is proposed, which transforms information into DNA readout via DNA‐tagged antibodies labels single unique cell barcodes. In 184‐electrode DMF‐Protein‐seq system, ≈1800 simultaneously detected per experimental run. The device harnessing low‐adsorbed hydrophobic surface contaminants‐isolated reaction space supports utilization (>90%) mapping reads input ranging from 140 to 2000. This system leverages split&pool strategy DMF chip first time overcome platform restriction analysis throughput replace traditionally tedious bench‐top combinatorial barcoding. With benefits efficiency sensitivity analysis, offers great potential classification drug monitoring expression at single‐cell level.

Язык: Английский

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Plasticity of BioPhi-Driven Humanness Optimization in ScFv-CD99 Binding Affinity Validated Through AlphaFold, HADDOCK, and MD Simulations

Computational and Structural Biotechnology Journal, Год журнала: 2025, Номер 27, С. 369 - 382

Опубликована: Янв. 1, 2025

BioPhi-guided humanization was utilized to enhance the humanness of a humanized single-chain variable fragment targeting CD99, leading development two variants: HuScFvMT99/3BP and HuScFvMT99/3HY. The variant incorporated framework region modifications, modest improvements in humanness, particularly VH domain, although VL domain remained suboptimal. To address this limitation, HuScFvMT99/3HY designed by combining wild-type with HuScFvMT99/3BP. Molecular dynamics simulations employing AlphaFold2, AlphaFold3, HADDOCK were performed evaluate HuScFv-CD99 peptide complexes. AF2-based demonstrated enhanced binding free energy (ΔGbinding) for both variants compared HuScFvMT99/3WT. However, ΔGbinding values obtained from AF3 HD inconsistent, exhibiting weakest affinity. While patterns derived AlphaFold3 aligned, amino acid decomposition analysis revealed variations interaction coordinates predicted Root-mean-square deviation indicated improved structural stability (0.975 Å) (1.075 relative HuScFvMT99/3WT (1.225 Å). Biolayer interferometry further confirmed that exhibited highest affinity (KD = 1.35 × 10⁻⁷ M) 2.64 3.95 M). Supporting evidence provided ELISA flow cytometry experiments. PITHA high immunogenicity risk all variants, despite displaying larger complementarity-determining (CDR) cavity, more hydrophobic CDR-H3 loop. These findings highlight delicate balance between enhancing preserving functional integrity critical therapeutic antibody development.

Язык: Английский

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Combating chemoresistance: Current approaches & nanocarrier mediated targeted delivery

Biochimica et Biophysica Acta (BBA) - Reviews on Cancer, Год журнала: 2025, Номер unknown, С. 189261 - 189261

Опубликована: Янв. 1, 2025

Язык: Английский

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