Targeting CRABP-II overcomes pancreatic cancer drug resistance by reversing lipid raft cholesterol accumulation and AKT survival signaling.
Targeting CRABP-II overcomes pancreatic cancer drug resistance by reversing lipid raft cholesterol accumulation and AKT survival signaling.
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DOI：
10.1186/s13046-022-02261-0
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发表时间：
2022-03-08
期刊：
影响因子：
--
通讯作者：
Zhou L
中科院分区：
文献类型：
--
作者：
Yu S;Wang L;Che D;Zhang M;Li M;Naito M;Xin W;Zhou L
对标准治疗的抵抗是胰腺导管腺癌（PDAC）预后不良的主要原因。因此，迫切需要开发新的治疗方法来克服PDAC耐药性。CRABP-II在所有PDAC中均呈高表达，而在正常胰腺组织和慢性胰腺炎中不表达。CRABP-II显示促进PDAC迁移和转移，但其在促进PDAC耐药性中的潜在作用尚不清楚。通过免疫组织化学评估人原发性和复发性PDAC组织的配对队列的CRABP-II表达。使用CRISPR/cas9基因编辑建立CRABP-II敲除细胞系，并进行MTT测定以评估体外吉西他滨敏感性。用切割型caspase-3/PARP杂交和Annexin V染色检测细胞凋亡。采用基因表达谱芯片、Q-PCR、Western blot、Co-IP和RNA-IP等方法研究CRABP-Ⅱ的分子功能。采用蔗糖梯度超离心法分离脂筏，LC-MS-MS法测定胆固醇含量。建立皮下CDX模型和原位PDX模型以检查SNIPER-11的功效以及SNIPER-11与吉西他滨之间的体内协同效应。复发PDAC组织中CRABP-II表达增高，与预后不良相关。吉西他滨耐药细胞系表现出增加的CRABP-II水平，而CRABP-II敲除使PDAC细胞对吉西他滨重新敏感。CRABP-II的异常表达通过与HuR的协同作用增加了SREBP-1c mRNA的稳定性，并上调了SREBP-1c的下游基因，从而有利于胆固醇在脂筏中的摄取和积累。增加脂筏胆固醇积累促进ATK存活信号传导和PDAC耐药性。小分子化合物SNIPER-11治疗有效地诱导CRABP-II蛋白降解，诱导细胞凋亡，并抑制肿瘤生长。SNIPER-11和吉西他滨的组合显著降低了CDX/PDX中的脂筏胆固醇含量，并显著抑制了肿瘤进展。这些发现将CRABP-II鉴定为胆固醇代谢的新型调节剂，并表明CRABP-II是克服PDAC耐药性的选择性靶点。在线版本包含补充材料，可通过10.1186/s13046-022-02261-0获得。
Resistance to standard therapy is a major reason for the poor prognosis of pancreatic ductal adenocarcinoma (PDAC). Developing novel therapy to overcome PDAC drug-resistance is urgently needed. CRABP-II was highly expressed in all PDAC but not expressed in normal pancreatic tissues and chronic pancreatitis. CRABP-II was shown to promote PDAC migration and metastasis while its potential role in promoting PDAC drug-resistance was not known. A paired cohort of human primary and relapsing PDAC tissues was assessed for CRABP-II expression by immunohistochemistry. CRISPR/cas9 gene editing was used to establish CRABP-II knockout cell lines and MTT assays were performed to assess gemcitabine sensitivity in vitro. Cleaved caspase-3/PARP blots and Annexin V staining were conducted to detect cell apoptosis. Gene expression microarray, Q-PCR, western blots, Co-IP and RNA-IP were used to study the molecular function of CRABP-II. Sucrose gradient ultracentrifugation was applied to isolate lipid rafts and LC–MS-MS was used to assess cholesterol content. Both subcutaneous CDX models and orthotopic PDX models were established to examine the efficacy of SNIPER-11 and the synergistic effect between SNIPER-11 and gemcitabine in vivo. A higher expression of CRABP-II was found in relapsing PDAC tissue and was associated with poor prognosis. Gemcitabine-resistant cell lines exhibited increased level of CRABP-II, while CRABP-II knockout resensitized PDAC cells to gemcitabine. Mechanistically, aberrant expression of CRABP-II increased the stability of SREBP-1c mRNA through cooperation with HuR and upregulated the downstream genes of SREBP-1c to favor cholesterol uptake and accumulation in lipid rafts. Increased lipid raft cholesterol accumulation facilitated ATK survival signaling and PDAC drug resistance. The small compound SNIPER-11 treatment effectively induced CRABP-II protein degradation, induced apoptosis, and suppressed tumor growth. Combination of SNIPER-11 and gemcitabine significantly reduced the lipid raft cholesterol content in CDX/PDX and profoundly inhibited tumor progression. These findings identified CRABP-II as a novel regulator of cholesterol metabolism and suggested that CRABP-II is a selective target for overcoming PDAC drug resistance. The online version contains supplementary material available at 10.1186/s13046-022-02261-0.
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影响因子：
6
作者：
Li, YC;Park, MJ;Kim, YN
通讯作者：
Kim, YN
影响因子：
5.4
作者：
Gu, Liang;Saha, Sourav Taru;Kaur, Mandeep
通讯作者：
Kaur, Mandeep
影响因子：
4.8
作者：
Dong, D;Ruuska, SE;Noy, N
通讯作者：
Noy, N
DOI：
10.1073/pnas.1005191107
发表时间：
2010-07-06
影响因子：
11.1
作者：
Marquart, Tyler J.;Allen, Ryan M.;Baldan, Angel
通讯作者：
Baldan, Angel
影响因子：
5.3
作者：
Dormoy-Raclet, Virginie;Menard, Isabelle;Gallouzi, Imed-Eddine
通讯作者：
Gallouzi, Imed-Eddine