FLASH effect on the liver and spleen of mice—a transcriptional perspective

IF 3.2 2区医学 Q1 RADIOLOGY, NUCLEAR MEDICINE & MEDICAL IMAGING

Medical physics Pub Date : 2026-10-21 DOI:10.1002/mp.70720

Yuqi Ma, Xinlin Zhu, Tianyu Yang, Jianfeng Lv, Xueqing Yan, Gen Yang, Xiaoji Lin, Zhifei Cao

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引用次数: 0

Abstract

Background

FLASH radiotherapy (FLASH-RT), delivered at an ultra-high dose rate (typically ≥ 40 Gy/s), has been shown to reduce normal tissue toxicity compared with conventional dose-rate radiotherapy (CONV-RT). However, the transcriptomic mechanisms underlying FLASH-mediated protection remain incompletely understood.

Purpose

This study aimed to compare the biological effects of FLASH and CONV irradiation on mouse liver and spleen tissues and to identify molecular pathways associated with the FLASH effect from a transcriptional perspective.

Methods

C57BL/6J male mice received a single 12 Gy abdominal dose delivered at either 2 Gy/min (CONV) or 100 Gy/s (FLASH). Twelve days after irradiation, spleen and liver tissues were harvested for histological section analysis and RNA-seq. Inflammatory responses in the spleen and liver were assessed by hematoxylin-eosin staining (H&E staining) and immunohistochemical quantification. A series of bioinformatic analyses, such as gene enrichment, protein–protein interaction (PPI) analysis, and transcription factor prediction, were performed to explore the effect of FLASH irradiation. Subsequently, reverse transcription quantitative PCR (RT-qPCR), western blot analysis, and immunohistochemical quantification were used to verify the bioinformatic results.

Results

Histological and immunohistochemical analyses showed less prominent inflammatory changes and lower immune-cell marker staining in mouse spleen and liver after FLASH than after CONV irradiation at 12 days post-irradiation. Bioinformatic analysis and corresponding experimental results revealed that compared to CONV irradiation, FLASH irradiation induced down-regulation of oxidative phosphorylation and cytoplasmic translation pathways in both mouse liver and spleen. Meanwhile, immune-related pathways, especially interferon β (IFNβ) signaling pathways, were significantly activated in both tissues after FLASH than after CONV irradiation. For these three pathways, key transcription factors were further identified and verified. Notably, STAT1 was found to play a potentially critical role in the upregulation of the IFNβ-related pathway under FLASH irradiation conditions in both liver and spleen.

Conclusions

FLASH irradiation induces distinct metabolic and immune adaptations in normal mouse liver and spleen tissues compared with CONV irradiation. The coordinated suppression of metabolic and translational pathways, together with activation of IFNβ-STAT1 signaling program, provides transcriptomic evidence for potential mechanisms underlying FLASH-mediated normal tissue sparing. These findings offer candidate pathways and molecular markers for future functional validation and support the clinical translation of FLASH-RT.

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FLASH对小鼠肝脏和脾脏的影响——一个转录的视角

与传统剂量率放疗（convr）相比，以超高剂量率（通常≥40 Gy/s）进行的FLASH放疗（FLASH- rt）已被证明可以降低正常组织毒性。然而，flash介导保护的转录组学机制仍然不完全清楚。本研究旨在比较FLASH和CONV辐照对小鼠肝脏和脾脏组织的生物学效应，并从转录角度确定与FLASH效应相关的分子通路。方法C57BL/6J雄性小鼠以2 Gy/min （CONV）或100 Gy/s （FLASH）给药12 Gy单次腹腔给药。照射12天后，取脾和肝组织进行组织切片分析和rna测序。采用苏木精-伊红染色（H&；E染色）和免疫组织化学定量法评估脾脏和肝脏的炎症反应。通过一系列的生物信息学分析，如基因富集、蛋白-蛋白相互作用（PPI）分析和转录因子预测，来探讨FLASH辐照的影响。随后，采用反转录定量PCR （RT-qPCR）、western blot分析和免疫组织化学定量分析对生物信息学结果进行验证。结果经组织学和免疫组织化学分析显示，与CONV照射后12 d相比，FLASH照射后小鼠脾脏和肝脏炎症变化不明显，免疫细胞标记物染色较低。生物信息学分析和相应的实验结果显示，与CONV照射相比，FLASH照射诱导小鼠肝脏和脾脏氧化磷酸化和细胞质翻译通路下调。与此同时，与CONV照射相比，FLASH照射后两种组织的免疫相关通路，尤其是干扰素β (IFNβ)信号通路明显被激活。对于这三条通路，我们进一步鉴定并验证了关键转录因子。值得注意的是，在肝脏和脾脏的FLASH照射条件下，STAT1在ifn β相关通路的上调中发挥了潜在的关键作用。结论与CONV照射相比，FLASH照射可诱导正常小鼠肝脏和脾脏组织的代谢和免疫适应。代谢和翻译途径的协同抑制，以及IFNβ-STAT1信号程序的激活，为flash介导的正常组织保留的潜在机制提供了转录组学证据。这些发现为未来功能验证提供了候选途径和分子标记，并支持FLASH-RT的临床翻译。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Medical physics 医学-核医学

CiteScore

6.80

自引率

15.80%

发文量

660

审稿时长

1.7 months

期刊介绍： Medical Physics publishes original, high impact physics, imaging science, and engineering research that advances patient diagnosis and therapy through contributions in 1) Basic science developments with high potential for clinical translation 2) Clinical applications of cutting edge engineering and physics innovations 3) Broadly applicable and innovative clinical physics developments Medical Physics is a journal of global scope and reach. By publishing in Medical Physics your research will reach an international, multidisciplinary audience including practicing medical physicists as well as physics- and engineering based translational scientists. We work closely with authors of promising articles to improve their quality.