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Identification and characterization of oncogenic KRAS G12V inhibitory peptides by phage display, molecular docking
Jirakrit Saetang1, Montarop Yamabhai2, Kuntalee Rangnoi2
1International Center of Excellence in Seafood Science and Innovation, Faculty of Agro-Industry, Prince of Songkla University, Hat Yai, Songkhla, 90110 Thailand.
Abstract:
The KRAS G12V mutation is a critical oncogenic driver in aggressive cancers, yet developing effective inhibitors remains challenging due to its elusive structural features. In this study, we employed phage display technology using both linear and cyclic peptide libraries to identify inhibitory peptides against KRAS G12V. Through subtractive bio-panning against wild-type KRAS, we identified two 23-mer peptides (Pep I and Pep II) that demonstrated selective binding to KRAS G12V. Molecular dynamics simulations revealed distinct binding mechanisms - Pep II showed stronger selective binding to G12V (-35.96 kcal/mol) compared to wild-type KRAS (-18.06 kcal/mol), while Pep I exhibited similar binding energies but interacted with different regions. Notably, Pep I bound to functional regions in KRAS G12V but non-functional regions in wild-type KRAS. Both peptides demonstrated significant inhibition of KRAS G12V-carrying cancer cell lines (NCI-H2444 and SW620), reducing cell viability by 70-75 % at 400 μM after 48 h while showing minimal effects (20-30 % reduction) on wild-type KRAS-carrying Caco-2 cells, which is equal to DMSO diluent control. These findings provide new insights into peptide-based targeting of KRAS G12V and demonstrate the potential of using subtractive phage display for developing selective inhibitors against oncogenic mutations.
Insights
Researchers developed peptide inhibitors targeting the KRAS G12V mutation using phage display. These peptides selectively inhibit KRAS G12V cancer cells, offering a promising new therapeutic strategy.
Area of Science:
- Oncology
- Molecular Biology
- Biotechnology
Background:
- The KRAS G12V mutation is a key driver in aggressive cancers, posing significant therapeutic challenges due to its complex structure.
- Targeting oncogenic mutations like KRAS G12V is crucial for developing effective cancer therapies.
Purpose of the Study:
- To identify and characterize novel peptide inhibitors targeting the KRAS G12V mutation using phage display technology.
- To investigate the binding mechanisms and selectivity of identified peptides against KRAS G12V and wild-type KRAS.
- To evaluate the efficacy of these peptides in inhibiting the proliferation of cancer cells harboring the KRAS G12V mutation.
Main Methods:
- Phage display technology with linear and cyclic peptide libraries was employed for inhibitor screening.
- Subtractive bio-panning against wild-type KRAS was performed to ensure target specificity.
- Molecular dynamics simulations were utilized to analyze peptide-protein interactions and binding affinities.
- In vitro assays were conducted to assess the impact of peptides on cancer cell viability.
Main Results:
- Two 23-mer peptides, Pep I and Pep II, were identified with selective binding to KRAS G12V.
- Molecular dynamics simulations indicated stronger selective binding for Pep II to KRAS G12V (-35.96 kcal/mol) compared to wild-type KRAS (-18.06 kcal/mol).
- Both peptides significantly reduced the viability of KRAS G12V-positive cancer cell lines (NCI-H2444, SW620) by 70-75% while showing minimal effects on wild-type KRAS-positive cells (Caco-2).
Conclusions:
- Subtractive phage display is an effective strategy for developing selective peptide inhibitors against oncogenic mutations like KRAS G12V.
- The identified peptides, Pep I and Pep II, demonstrate significant potential as therapeutic agents for KRAS G12V-driven cancers.
- These findings offer new insights into peptide-based targeting of KRAS G12V and pave the way for further drug development.
