TP53 R249S mutation in hepatic organoids captures the predisposing cancer risk.
TP53 R249S mutation in hepatic organoids captures the predisposing cancer risk.
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肝癌中TP53 R249S突变捕获了易感性的癌症风险。
DOI：
10.1002/hep.32802
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发表时间：
2023-09-01
期刊：
影响因子：
13.5
通讯作者：
Wong, Nathalie
中科院分区：
文献类型：
--
作者：
Lam, Yin Kau;Yu, Jianqing;Huang, Hao;Ding, Xiaofan;Wong, Alissa M.;Leung, Howard H.;Chan, Anthony W.;Ng, Kelvin K.;Xu, Mingjing;Wang, Xin;Wong, Nathalie
肝细胞癌（HCC）的主要基因组驱动因素目前已得到充分认可，但建立其在人类HCC起始中的作用的模型仍然很少。在这里，我们在实验系统中使用人类肝脏类器官来模拟人类肝癌发生的早期阶段，即TP 53缺失和L3环R249 S突变的遗传病变。此外，HCC细胞系的染色质免疫沉淀测序（ChIP-seq）揭示了重要的功能性见解，即HCC的起始是由于TP 53缺陷导致的肿瘤抑制功能丧失和突变型p53的功能获得活动。人肝类器官由手术非肿瘤肝组织产生。在肝类器官中TP 53的CRISPR敲除一致地显示出肿瘤样形态学变化、干性增加和不受限制的体外增殖。为了概括TP 53在人HCC中的状态，我们在TP 53敲除的类器官中过表达突变体R249 S。在异种移植中观察到自发的致瘤潜力和真正的HCC组织学的增加。HCC细胞系的ChIP-seq分析强调了L3环p53突变体在染色质重塑和克服外源性应激中的功能获得特性。更重要的是，突变体R249 S对PSMF 1的直接转录激活可以增加类器官对内质网应激的抗性，这在拯救实验中很容易被PSMF 1敲低所消除。在原发性肝癌肿瘤和基因组编辑的肝脏类器官的患者队列中，定量聚合酶链反应证实了ChIP-seq结果，并验证了受L3突变体调节的优先基因，特别是那些富含R249 S的基因。我们发现TP 53缺失和L3突变具有不同的致瘤作用，这两种突变共同赋予正常肝细胞早期克隆优势和促生存功能。
Major genomic drivers of hepatocellular carcinoma (HCC) are nowadays well recognized, although models to establish their roles in human HCC initiation remain scarce. Here, we used human liver organoids in experimental systems to mimic the early stages of human liver carcinogenesis from the genetic lesions of TP53 loss and L3 loop R249S mutation. In addition, chromatin immunoprecipitation sequencing (ChIP‐seq) of HCC cell lines shed important functional insights into the initiation of HCC consequential to the loss of tumor‐suppressive function from TP53 deficiency and gain‐of‐function activities from mutant p53. Human liver organoids were generated from surgical nontumor liver tissues. CRISPR knockout of TP53 in liver organoids consistently demonstrated tumor‐like morphological changes, increased in stemness and unrestricted in vitro propagation. To recapitulate TP53 status in human HCC, we overexpressed mutant R249S in TP53 knockout organoids. A spontaneous increase in tumorigenic potentials and bona fide HCC histology in xenotransplantations were observed. ChIP‐seq analysis of HCC cell lines underscored gain‐of‐function properties from L3 loop p53 mutants in chromatin remodeling and overcoming extrinsic stress. More importantly, direct transcriptional activation of PSMF1 by mutant R249S could increase organoid resistance to endoplasmic reticulum stress, which was readily abrogated by PSMF1 knockdown in rescue experiments. In a patient cohort of primary HCC tumors and genome‐edited liver organoids, quantitative polymerase chain reaction corroborated ChIP‐seq findings and verified preferential genes modulated by L3 mutants, especially those enriched by R249S. We showed differential tumorigenic effects from TP53 loss and L3 mutations, which together confer normal hepatocytes with early clonal advantages and prosurvival functions.
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影响因子：
29.4
作者：
Laurent-Puig, P;Legoix, P;Zucman-Rossi, J
通讯作者：
Zucman-Rossi, J
影响因子：
50.3
作者：
Ghosh M;Saha S;Bettke J;Nagar R;Parrales A;Iwakuma T;van der Velden AWM;Martinez LA
通讯作者：
Martinez LA
影响因子：
--
作者：
Liu, Xiaoying;Green, Richard M
通讯作者：
Green, Richard M
影响因子：
8
作者：
Dong, P.;Karaayvaz, M.;Jia, N.;Kaneuchi, M.;Hamada, J.;Watari, H.;Sudo, S.;Ju, J.;Sakuragi, N.
通讯作者：
Sakuragi, N.
DOI：
10.1073/pnas.0607286103
发表时间：
2006-10-10
影响因子：
11.1
作者：
Joerger, Andreas C.;Ang, Hwee Ching;Fersht, Alan R.
通讯作者：
Fersht, Alan R.