ROS1重排是非小细胞肺癌(NSCLC)中的一个关键治疗靶点。然而，目前对ROS1融合分子模式的综合分析相对有限。本研究旨在使用下一代测序技术(NGS)对134名中国NSCLC患者中的135个ROS1融合进行分析，从而填补这一空白。根据其发生率，这些融合被分为常见和不常见两类。我们的研究首次揭示了ROS1内部断裂位点的独特分布偏好，常见融合发生在31-33内含子，而不常见融合发生在34和35内含子。此外，我们在ROS1的内部第28内含子中发现了先前未知的断裂位点。此外，我们还在ROS1的融合伴侣和断裂位点的分布模式之间发现了密切的关联，为我们对ROS1融合的分子景观提供了重要见解。我们还通过严格的验证方法证实了ROS1融合在DNA基于下一代测序(DNA NGS)和RNA基于下一代测序(RNA NGS)之间存在不一致的断裂位点。这些不一致性归因于不同的剪接导致ROS1融合在外框或外显子中。这些发现对我们理解ROS1融合的分子特征具有重要意义，对面板设计以及具有ROS1重排的NSCLC患者的治疗有重要影响。本文受版权保护，版权所有。

ROS proto-oncogene 1, receptor tyrosine kinase (ROS1) rearrangements are a crucial therapeutic target in non-small cell lung cancer (NSCLC). However, there is limited comprehensive analysis of the molecular patterns of ROS1 fusions. This study aimed to address this gap by analyzing 135 ROS1 fusions from 134 Chinese NSCLC patients using next-generation sequencing (NGS). The fusions were categorized into common and uncommon based on their incidence. Our study revealed, for the first time, a unique distribution preference of breakpoints within ROS1, with common fusions occurring in introns 31-33 and uncommon fusions occurring in introns 34 and 35. Additionally, we identified previously unknown breakpoints within intron 28 of ROS1. Furthermore, we identified a close association between the distribution patterns of fusion partners and breakpoints on ROS1, providing important insights into the molecular landscape of ROS1 fusions. We also confirmed the presence of inconsistent breakpoints in ROS1 fusions between DNA-based next-generation sequencing (DNA NGS) and RNA-based NGS (RNA NGS) through rigorous validation methods. These inconsistencies were attributed to alternative splicing resulting in out-of-frame or exonic ROS1 fusions. These findings significantly contribute to our understanding of the molecular characteristics of ROS1 fusions, which have implications for panel design and the treatment of NSCLC patients with ROS1 rearrangements.This article is protected by copyright. All rights reserved.