Related Experiment Video
Updated: Jan 17, 2026
Author Spotlight: Advancements in Molecular Biomarker Testing for Non-Squamous Non-Small Cell Lung Cancer
Published on: September 8, 2023
Preclinical Prediction of Resistance Mutations and Proposal of Sequential Treatment Strategies for ALK-positive Lung
Yuki Takei1, Hirotaka Kuroiwa1, Chisaki Arai1
1Department of Life Science and Medical Bioscience, Graduate School of Advanced Science and Engineering, Waseda University, 2‑2 Wakamatsu‑cho, Shinjuku‑ku, Tokyo, 162‑0056, Japan.
Background:
Anaplastic lymphoma kinase (ALK) gene rearrangements occur in approximately 5% of non-small cell lung cancers (NSCLCs). Although ALK tyrosine kinase inhibitors provide substantial clinical benefits, acquired resistance-conferring mutations frequently emerge, leading to disease progression. Preclinical prediction of these mutations might help guide the development of more effective sequential treatment strategies prior to clinical application.
Objective:
To predict the emergence of resistance mutations to the investigational ALK inhibitors zotizalkib (TPX-0131), gilteritinib (ASP2215), and neladalkib (NVL-655) following resistance to first-line alectinib and assess the potential of these drugs as second-line therapies.
Methods:
A polymerase chain reaction (PCR)-based mutagenesis system was used to introduce random mutations into ALK cDNA harboring representative alectinib-resistant mutations. Mutant libraries were expressed in Ba/F3 cells, which were exposed to each inhibitor. Drug-resistant clones were isolated, sequenced, and evaluated for drug sensitivity using viability assays and immunoblotting.
Results:
Several resistance mutations against zotizalkib, gilteritinib, and neladalkib were identified. Sequential use of these agents effectively suppressed all predicted resistance patterns with G1202R or I1171N.
Conclusions:
This PCR-based platform provides a valuable approach for anticipating resistance mutations and guiding the design of optimized sequential therapies. Zotizalkib, gilteritinib, and neladalkib might represent promising alternatives to lorlatinib as second-line treatments for ALK-positive NSCLC.
Key Points:
• A PCR-based mutation prediction system was successfully applied to fourth-generation ALK inhibitors. • Neladalkib showed efficacy against G1202R-positive relapses with minimal evidence of secondary resistance mutations. • Sequential combinations of gilteritinib with either neladalkib or ensartinib may sustain efficacy and delay resistance in I1171N-positive relapses.
Insights
Predicting Anaplastic Lymphoma Kinase (ALK) resistance mutations in non-small cell lung cancer (NSCLC) is crucial. A PCR-based system identified mutations, guiding sequential therapies with novel ALK inhibitors like zotizalkib, gilteritinib, and neladalkib.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Anaplastic Lymphoma Kinase (ALK) gene rearrangements are present in about 5% of non-small cell lung cancers (NSCLCs).
- Acquired resistance mutations to ALK tyrosine kinase inhibitors frequently emerge, leading to disease progression.
- Preclinical prediction of resistance mutations can inform sequential treatment strategies.
Purpose of the Study:
- To predict resistance mutations to investigational ALK inhibitors zotizalkib, gilteritinib, and neladalkib after resistance to first-line alectinib.
- To assess the potential of these drugs as second-line therapies for ALK-positive NSCLC.
Main Methods:
- A polymerase chain reaction (PCR)-based mutagenesis system was employed to introduce random mutations into ALK cDNA.
- Mutant libraries were expressed in Ba/F3 cells and exposed to zotizalkib, gilteritinib, and neladalkib.
- Drug-resistant clones were isolated, sequenced, and evaluated for drug sensitivity.
Main Results:
- Several resistance mutations against zotizalkib, gilteritinib, and neladalkib were identified.
- The sequential use of these agents effectively suppressed predicted resistance patterns, including G1202R or I1171N mutations.
- Neladalkib demonstrated efficacy against G1202R-positive relapses with minimal secondary resistance.
- Sequential combinations of gilteritinib with neladalkib or ensartinib showed potential for sustained efficacy in I1171N-positive relapses.
Conclusions:
- A PCR-based platform effectively anticipates ALK resistance mutations, aiding in the design of optimized sequential therapies.
- Zotizalkib, gilteritinib, and neladalkib show promise as second-line treatments for ALK-positive NSCLC, potentially offering alternatives to lorlatinib.
- This approach facilitates the development of next-generation ALK inhibitors and treatment strategies.
More Related Videos
05:17Detection of Targetable Alterations in Non-small Cell Lung Cancer using Next-generation Sequencing
Published on: October 10, 2025
08:46Implementation of In Vitro Drug Resistance Assays: Maximizing the Potential for Uncovering Clinically Relevant Resistance Mechanisms
Published on: December 9, 2015
Related Concept Videos
02:50Combination Therapies and Personalized Medicine
The combination of the drug acetazolamide and sulforaphane is a good example of combination therapy to treat cancer. The cells in the interior of a large tumor often die due to the hypoxic and...
02:57Targeted Cancer Therapies
There are several types of targeted therapies against...
02:56Treatment Resistant Cancers