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Updated: Mar 12, 2026
Identification of RNAs Engaged in Direct RNA-RNA Interaction with a Long Non-Coding RNA
Published on: July 9, 2021
Coupling an EML4-ALK-centric interactome with RNA interference identifies sensitizers to ALK inhibitors
Guolin Zhang1, Hannah Scarborough2, Jihye Kim3
1Department of Thoracic Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA.
Abstract:
Patients with lung cancers harboring anaplastic lymphoma kinase (ALK) gene fusions benefit from treatment with ALK inhibitors, but acquired resistance inevitably arises. A better understanding of proximal ALK signaling mechanisms may identify sensitizers to ALK inhibitors that disrupt the balance between prosurvival and proapoptotic effector signals. Using affinity purification coupled with mass spectrometry in an ALK fusion lung cancer cell line (H3122), we generated an ALK signaling network and investigated signaling activity using tyrosine phosphoproteomics. We identified a network of 464 proteins composed of subnetworks with differential response to ALK inhibitors. A small hairpin RNA screen targeting 407 proteins in this network revealed 64 and 9 proteins that when knocked down sensitized cells to crizotinib and alectinib, respectively. Among these, knocking down fibroblast growth factor receptor substrate 2 (FRS2) or coiled-coil and C2 domain-containing protein 1A (CC2D1A), both scaffolding proteins, sensitized multiple ALK fusion cell lines to the ALK inhibitors crizotinib and alectinib. Collectively, our data set provides a resource that enhances our understanding of signaling and drug resistance networks consequent to ALK fusions and identifies potential targets to improve the efficacy of ALK inhibitors in patients.
Insights
Discovering new ways to overcome drug resistance in anaplastic lymphoma kinase (ALK) fusion lung cancer is crucial. This study identifies key proteins, like FRS2 and CC2D1A, that sensitize cancer cells to ALK inhibitors, offering hope for improved patient outcomes.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Anaplastic lymphoma kinase (ALK) gene fusions drive certain lung cancers.
- Patients benefit from ALK inhibitors, but drug resistance is a significant challenge.
- Understanding proximal ALK signaling is key to overcoming resistance.
Purpose of the Study:
- To identify novel targets that sensitize ALK-fusion lung cancer to ALK inhibitors.
- To elucidate the signaling networks involved in ALK-driven lung cancer and resistance.
Main Methods:
- Utilized affinity purification coupled with mass spectrometry to generate an ALK signaling network in an ALK-fusion lung cancer cell line.
- Employed tyrosine phosphoproteomics to investigate signaling activity.
- Performed a small hairpin RNA screen on 407 proteins within the network.
Main Results:
- Identified a comprehensive ALK signaling network of 464 proteins.
- Discovered subnetworks with differential responses to ALK inhibitors.
- Knockdown of fibroblast growth factor receptor substrate 2 (FRS2) and coiled-coil and C2 domain-containing protein 1A (CC2D1A) sensitized multiple ALK-fusion cell lines to crizotinib and alectinib.
Conclusions:
- FRS2 and CC2D1A are potential therapeutic targets to enhance ALK inhibitor efficacy.
- The generated signaling network provides a valuable resource for understanding ALK-driven lung cancer and resistance.
- This research identifies strategies to improve treatment outcomes for patients with ALK-fusion lung cancer.
