Related Experiment Video
Updated: May 3, 2026
Bimolecular Fluorescence Complementation
Published on: April 15, 2011
Integrative Multi-Omics Approaches Reveal Selectivity Profiles and Molecular Mechanisms of FIIN-2, a Covalent FGFR
Ying Fu1, Dandan Zhu1, Xiaojuan Chen1
1Department of Oncology, NHC Key Laboratory of Cancer Proteomics and State Local Joint Engineering Laboratory for Anticancer Drugs, National Clinical Research Center for Geriatric Disorders, Xiangya Hospital, Central South University, Changsha, Hunan, 410008, China.
Abstract:
Fibroblast growth factor receptor (FGFR) inhibitors are emerged as an important class of targeted therapies in oncology, targeting key pathways associated with tumor growth, angiogenesis, and resistance to conventional treatments. FIIN-2, the first irreversible covalent pan-FGFR inhibitor, has shown promise in overcoming resistance due to gatekeeper mutations; however, its selectivity and molecular mechanisms in tumors remain poorly understood. In this study, an FIIN-2 chemical probe is designed and synthesized to identify both established and novel targets in hepatocellular carcinoma (HCC) via chemoproteomic profiling. An integrative multi-omics approach, including chemoproteomic, phosphoproteomic, transcriptomic, and proteomic analyses, is utilized to elucidate the full spectrum of target proteins, signaling pathways, and downstream effectors regulated by FIIN-2 in HCC. Notably, adenosine monophosphate-activated protein kinase α1 (AMPKα1) is identified as a novel target of FIIN-2, with Cys185 identified as its covalent binding site. These findings reveal that FIIN-2 can induce autophagy by directly binding to and activating AMPKα1, influencing its anti-tumor activity in HCC cells. Overall, this study greatly advances the understanding of FIIN-2's on- and off-target effects, offering a comprehensive view of its molecular mechanisms in cancer cells. The integrative multi-omics approach provides a valuable framework for the development and optimization of covalent kinase inhibitors.
Insights
FIIN-2, a covalent fibroblast growth factor receptor (FGFR) inhibitor, activates AMPKα1, a novel target, to induce autophagy and exert anti-tumor effects in hepatocellular carcinoma (HCC). This study clarifies FIIN-2
Area of Science:
- Oncology
- Molecular Biology
- Biochemistry
Background:
- Fibroblast growth factor receptor (FGFR) inhibitors are crucial targeted therapies in oncology.
- FIIN-2 is the first irreversible covalent pan-FGFR inhibitor, showing potential against gatekeeper mutations.
- Understanding FIIN-2's selectivity and molecular mechanisms in tumors is limited.
Purpose of the Study:
- To design and synthesize an FIIN-2 chemical probe for target identification in hepatocellular carcinoma (HCC).
- To elucidate the full spectrum of FIIN-2's target proteins, signaling pathways, and downstream effectors using multi-omics.
- To investigate the molecular mechanisms underlying FIIN-2's anti-tumor activity in HCC.
Main Methods:
- Chemoproteomic profiling using an FIIN-2 chemical probe.
- Integrative multi-omics approach: chemoproteomic, phosphoproteomic, transcriptomic, and proteomic analyses.
- Identification of direct targets and binding sites through biochemical assays.
Main Results:
- Adenosine monophosphate-activated protein kinase α1 (AMPKα1) was identified as a novel target of FIIN-2.
- Cys185 on AMPKα1 was identified as the covalent binding site for FIIN-2.
- FIIN-2 directly binds and activates AMPKα1, leading to autophagy induction in HCC cells.
Conclusions:
- FIIN-2's novel target, AMPKα1, and its covalent binding site (Cys185) were elucidated.
- FIIN-2 induces autophagy via AMPKα1 activation, contributing to its anti-tumor effects in HCC.
- This study provides a comprehensive understanding of FIIN-2's molecular mechanisms and a framework for covalent kinase inhibitor development.
