GNAS(R201C) Induces Pancreatic Cystic Neoplasms in Mice That Express Activated KRAS by Inhibiting YAP1 Signaling.
GNAS(R201C) Induces Pancreatic Cystic Neoplasms in Mice That Express Activated KRAS by Inhibiting YAP1 Signaling.
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DOI：
10.1053/j.gastro.2018.08.006
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发表时间：
2018-11
期刊：
影响因子：
29.4
通讯作者：
Maitra A
中科院分区：
文献类型：
--
作者：
Ideno N;Yamaguchi H;Ghosh B;Gupta S;Okumura T;Steffen DJ;Fisher CG;Wood LD;Singhi AD;Nakamura M;Gutkind JS;Maitra A
编码刺激性G蛋白α亚基的GNAS热点突变在约60%的胰腺导管内乳头状黏液性肿瘤（IPMNs）中被检测到。我们构建了由KRAS诱导且在胰腺中也表达组成性激活形式的GNAS的IPMNs小鼠，并研究了肿瘤的发展。
我们构建了p48 - Cre；LSL - KrasG12D；Rosa26R - LSL - rtTA - TetO - GnasR201C小鼠（Kras；Gnas小鼠）；这些小鼠的胰腺组织在给予强力霉素时表达激活的KRAS，也表达一种突变形式的GNAS（GNASR201C）。未给予强力霉素的小鼠用作对照，并通过卡普兰 - 迈耶分析比较生存时间。在给予强力霉素后的不同时间点收集胰腺组织，并通过组织学进行分析。从小鼠中分离出胰腺导管腺癌（PDACs）并用于建立细胞系，通过逆转录PCR、免疫印迹、免疫组织化学以及集落形成和侵袭实验对细胞系进行分析。YAP的全长形式和突变形式在PDAC细胞中表达。从13例接受手术的IPMN患者中获取IPMN标本，并通过免疫组织化学进行分析。
所有Kras；Gnas小鼠都形成了类似于人类IPMNs的胰腺囊性病变；上皮异型增生的级别随时间增加。对照小鼠均未形成囊性病变。大约三分之一的Kras；Gnas小鼠在给予强力霉素后中位数为30周时发展为PDACs，而33%的对照小鼠发展为PDACs。与对照小鼠相比，GNASR201C的表达并未加速PDACs的发展。然而，在Kras；Gnas小鼠中观察到的肿瘤比对照小鼠更分化，并且表达更多与导管表型相关的基因。从Kras；Gnas小鼠中分离出的PDACs具有Hippo通路的激活；在这些肿瘤的细胞中，磷酸化的YAP1被隔离在细胞质中——这在具有GNAS突变的人类IPMNs中也被观察到。在对照小鼠的PDAC细胞中未观察到YAP1的隔离。
在胰腺中表达激活的KRAS的小鼠中，我们发现GNASR201C的表达导致更分化的肿瘤的发展，具有与导管表型相关的基因表达模式。突变的GNAS的表达导致磷酸化的YAP1被隔离在细胞质中，改变了肿瘤的进展。
Mutations at hot spots in GNAS, which encodes stimulatory G-protein, alpha subunits, are detected in ~60% of intraductal papillary mucinous neoplasms (IPMNs) of the pancreas. We generated mice with KRAS-induced IPMNs that also express a constitutively active form of GNAS in pancreas and studied tumor development. We generated p48-Cre; LSL-KrasG12D; Rosa26R-LSL-rtTA-TetO-GnasR201C mice (Kras;Gnas mice); pancreatic tissues of these mice express activated KRAS and also express a mutant form of GNAS (GNASR201C) upon doxycycline administration. Mice that were not given doxycycline were used as controls and survival times were compared by Kaplan-Meier analysis. Pancreasta were collected at different timepoints after doxycycline administration and analyzed by histology. Pancreatic ductal adenocarcinomas (PDACs) were isolated from mice and used to generate cell lines, which were analyzed by reverse transcription PCR, immunoblotting, immunohistochemistry, and colony formation and invasion assays. Full-length and mutant forms of YAP were expressed in PDAC cells. IPMN specimens were obtained from 13 patients with IPMN undergoing surgery and analyzed by immunohistochemistry. All Kras;Gnas mice developed pancreatic cystic lesions that resemble human IPMNs; the grade of epithelial dysplasia increased with time. None of the control mice developed cystic lesions. Approximately one-third of Kras;Gnas mice developed PDACs, at a median of 30 weeks after doxycycline administration, whereas 33% of control mice developed PDACs. Expression of GNASR201C did not accelerate the development of PDACs, compared with control mice. However, the neoplasms observed in Kras;Gnas mice were more differentiated, and expressed more genes associated with ductal phenotypes, than in control mice. PDACs isolated from Kras;Gnas mice had activation of the Hippo pathway; in cells from these tumors, phosphorylated YAP1 was sequestered in the cytoplasm—this was also observed in human IPMNs with GNAS mutations. Sequestration of YAP1 was not observed in PDAC cells from control mice. In mice that express activated KRAS in the pancreas, we found expression of GNASR201C to cause development of more differentiated tumors, with gene expression pattern associated with the ductal phenotype. Expression of mutant GNAS caused phosphorylated YAP1 to be sequestered in the cytoplasm, altering tumor progression.
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影响因子：
21.3
作者：
通讯作者：
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影响因子：
8
作者：
Murakami S;Shahbazian D;Surana R;Zhang W;Chen H;Graham GT;White SM;Weiner LM;Yi C
通讯作者：
Yi C
河马效应子YAP促进了对RAF和MEK靶向的癌症疗法的耐药性。
影响因子：
30.8
作者：
Lin L;Sabnis AJ;Chan E;Olivas V;Cade L;Pazarentzos E;Asthana S;Neel D;Yan JJ;Lu X;Pham L;Wang MM;Karachaliou N;Cao MG;Manzano JL;Ramirez JL;Torres JM;Buttitta F;Rudin CM;Collisson EA;Algazi A;Robinson E;Osman I;Muñoz-Couselo E;Cortes J;Frederick DT;Cooper ZA;McMahon M;Marchetti A;Rosell R;Flaherty KT;Wargo JA;Bivona TG
通讯作者：
Bivona TG
影响因子：
7.3
作者：
Amato, Eliana;dal Molin, Marco;Mafficini, Andrea;Yu, Jun;Malleo, Giuseppe;Rusev, Borislav;Fassan, Matteo;Antonello, Davide;Sadakari, Yoshihiko;Castelli, Paola;Zamboni, Giuseppe;Maitra, Anirban;Salvia, Roberto;Hruban, Ralph H.;Bassi, Claudio;Capelli, Paola;Lawlor, Rita T.;Goggins, Michael;Scarpa, Aldo
通讯作者：
Scarpa, Aldo
影响因子：
21.3
作者：
通讯作者：
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