p53突变偏向鳞状柱状交界区前体细胞向不典型增生而非柱状上皮化转变在Barrett食管中的作用

p53 mutation biases squamocolumnar junction progenitor cells towards dysplasia rather than metaplasia in Barrett's oesophagus

GUT

影响因子:25.8

分区:医学1区 Top / 胃肠肝病学1区

发表日期:2025 Jan 17

作者: Guodong Lian, Ermanno Malagola, Chengguo Wei, Qiongyu Shi, Junfei Zhao, Masahiro Hata, Hiroki Kobayashi, Yosuke Ochiai, Biyun Zheng, Xiaofei Zhi, Feijing Wu, Ruhong Tu, Osmel Companioni Nápoles, Wenjing Su, Leping Li, Changqing Jing, Man Chen, Leah Zamechek, Richard Friedman, Karol Nowicki-Osuch, Michael Quante, Jianwen Que, Timothy C Wang

DOI: 10.1136/gutjnl-2024-332095

摘要

p53突变在Barrett食管（BE）向食管腺癌（EAC）进展中早期发生，但其在胃底腺干细胞中的作用尚不清楚。本研究旨在探讨p53突变对BE向EAC进展中胃底腺前体细胞命运与功能的影响，特别是在慢性损伤压力下的变化。我们采用携带Trp53突变（R172H）的BE小鼠模型（L2-IL1β）研究p53对Cck2r+胃底腺前体细胞的作用。通过谱系追踪、病理分析、器官培养、单细胞RNA测序（scRNA-seq）及计算分析，观察前体细胞行为变化、分化模式及肿瘤进展。另外，进行分选的化生和突变前体细胞的原位移植，以评估其体内的肿瘤形成潜能。结果显示：p53突变在慢性损伤条件下起到开关作用，扩增前体细胞并抑制其向化生的分化。在L2-IL1β小鼠中，p53突变促进前体细胞扩增及谱系追踪，表现为从化生向不典型增生的转变。scRNA-seq分析显示，不典型增生细胞直接源自突变前体细胞，而非经过化生过程。在体外实验中，p53突变增强BE前体细胞的器官体形成效率、增殖能力、DNA损伤抗性及向非整倍体的进展。分选的化生细胞生长较差，未向不典型增生转变，而突变前体细胞经原位移植后形成不典型增生。计算分析表明，p53突变通过激活Notch信号抑制干细胞的分化。综上，p53突变通过增加未分化胃底腺前体细胞的扩增和适应性，阻止其向化生分化，促进BE的进展。

Abstract

While p53 mutations occur early in Barrett's oesophagus (BE) progression to oesophageal adenocarcinoma (EAC), their role in gastric cardia stem cells remains unclear.This study investigates the impact of p53 mutation on the fate and function of cardia progenitor cells in BE to EAC progression, particularly under the duress of chronic injury.We used a BE mouse model (L2-IL1β) harbouring a Trp53 mutation (R172H) to study the effects of p53 on Cck2r+ cardia progenitor cells. We employed lineage tracing, pathological analysis, organoid cultures, single-cell RNA sequencing (scRNA-seq) and computational analyses to investigate changes in progenitor cell behaviour, differentiation patterns and tumour progression. Additionally, we performed orthotopic transplantation of sorted metaplastic and mutant progenitor cells to assess their tumourigenic potential in vivo.The p53 mutation acts as a switch to expand progenitor cells and inhibit their differentiation towards metaplasia, but only amidst chronic injury. In L2-IL1β mice, p53 mutation increased progenitors expansion and lineage-tracing with a shift from metaplasia to dysplasia. scRNA-seq revealed dysplastic cells arise directly from mutant progenitors rather than progressing through metaplasia. In vitro, p53 mutation enhanced BE progenitors' organoid-forming efficiency, growth, DNA damage resistance and progression to aneuploidy. Sorted metaplastic cells grew poorly with no progression to dysplasia, while mutant progenitors gave rise to dysplasia in orthotopic transplantation. Computational analyses indicated that p53 mutation inhibited stem cell differentiation through Notch activation.p53 mutation contributes to BE progression by increasing expansion and fitness of undifferentiated cardia progenitors and preventing their differentiation towards metaplasia.