Therapeutic Targeting of TFE3/IRS-1/PI3K/mTOR Axis in Translocation Renal Cell Carcinoma.
Therapeutic Targeting of TFE3/IRS-1/PI3K/mTOR Axis in Translocation Renal Cell Carcinoma.
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DOI：
10.1158/1078-0432.ccr-18-0269
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发表时间：
2018-12-01
期刊：
影响因子：
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通讯作者：
Pili R
中科院分区：
文献类型：
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作者：
Damayanti NP;Budka JA;Khella HWZ;Ferris MW;Ku SY;Kauffman E;Wood AC;Ahmed K;Chintala VN;Adelaiye-Ogala R;Elbanna M;Orillion A;Chintala S;Kao C;Linehan WM;Yousef GM;Hollenhorst PC;Pili R
易位肾细胞癌（tRCC）是一种罕见的肾癌亚型，与各种TFE 3、TFEB或MITF基因融合相关，对RCC的标准治疗无反应。因此，新的治疗靶点的鉴定代表了这种疾病的未满足的需求。我们建立并鉴定了tRCC患者来源的异种移植物（PDX）RP-R 07，作为新的临床前药物开发模型，通过使用下一代测序和生物信息学分析，然后使用体外和体内模型评估抑制所鉴定的通路的治疗潜力。通过RNA-seq鉴定并通过RT-PCR验证具有染色体断裂点的SFPQ-TFE 3融合体（t（X;1）（p11.2; p34））的存在。TFE 3染色质免疫沉淀，然后进行深度测序（ChIP-Seq）分析，表明PI 3 K/AKT/mTOR途径的强烈富集。一致地，microRNA微阵列分析也鉴定了PI 3 K/AKT/mTOR作为RP-R 07中高度富集的途径。在另外的TFE 3-tRCC模型中PI 3/AKT/mTOR途径的上调通过与透明细胞RCC细胞相比在建立的tRCC细胞系中显著更高的磷酸-S6（P<0.0001）和磷酸-4EBP 1（P<0.0001）表达来证实。与单淋巴结抑制相比，同时垂直靶向PI 3 K/AKT和mTOR轴提供了更大的体外（P<0.0001）和体内（P<0.01）抗增殖作用。RP-R 07中TFE 3的敲低导致IRS-1的表达降低并抑制细胞增殖。这些结果将TFE 3/IRS-1/PI 3 K/AKT/mTOR鉴定为TFE 3-tRCC中潜在的失调途径，并表明通过使用双重PI 3 K/mTOR抑制剂对TFE 3-tRCC患者垂直抑制该轴的治疗潜力。
Translocation renal cell carcinoma (tRCC) represents a rare subtype of kidney cancer associated with various TFE3, TFEB, or MITF gene fusions that is not responsive to standard treatments for RCC. Therefore, the identification of new therapeutic targets represents an unmet need for this disease. We have established and characterized a tRCC patient-derived xenograft (PDX), RP-R07, as a novel preclinical model for drug development by using next generation sequencing and bioinformatics analysis.We then assessed the therapeutic potential of inhibiting the identified pathway using in vitro and in vivo models. The presence of a SFPQ-TFE3 fusion (t(X;1) (p11.2; p34)) with chromosomal break-points was identified by RNA-seq and validated by RT-PCR. TFE3 chromatin immunoprecipitation followed by deep sequencing (ChIP-Seq) analysis indicated a strong enrichment for the PI3K/AKT/mTOR pathway. Consistently, microRNA microarray analysis also identified PI3K/AKT/mTOR as a highly enriched pathway in RP-R07. Upregulation of PI3/AKT/mTOR pathway in additional TFE3-tRCC models were confirmed by significantly higher expression of phospho-S6 (P<0.0001) and phospho-4EBP1 (P<0.0001) in established tRCC cell lines compared to clear cell RCC cells. Simultaneous vertical targeting of both PI3K/AKT and mTOR axis provided a greater anti-proliferative effect both in vitro (P<0.0001) and in vivo (P<0.01) compared to single node inhibition. Knockdown of TFE3 in RP-R07 resulted in decreased expression of IRS-1 and inhibited cell proliferation. These results identify TFE3/IRS-1/PI3K/AKT/mTOR as a potential dysregulated pathway in TFE3-tRCC, and suggest a therapeutic potential of vertical inhibition of this axis by using a dual PI3K/mTOR inhibitor for TFE3-tRCC patients.