Transcriptomic changes underlying EGFR inhibitor resistance in human and mouse models of basal-like breast cancer.
Transcriptomic changes underlying EGFR inhibitor resistance in human and mouse models of basal-like breast cancer.
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DOI：
10.1038/s41598-022-25541-3
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发表时间：
2022-12-08
影响因子：
4.6
通讯作者：
Harrell JC
中科院分区：
文献类型：
--
作者：
Rashid NS;Boyd DC;Olex AL;Grible JM;Duong AK;Alzubi MA;Altman JE;Leftwich TJ;Valentine AD;Hairr NS;Zboril EK;Smith TM Jr;Pfefferle AD;Dozmorov MG;Harrell JC
本研究的目的是鉴定基底样乳腺癌细胞在对表皮生长因子受体抑制剂（EGFRi）产生耐药性过程中出现的转录组学变化，并鉴定一旦发生EGFRi耐药性就具有细胞毒性的药物。人患者来源的异种移植物（PDX）在免疫缺陷小鼠中生长，并用一组EGFRi治疗;选择EGFRi厄洛替尼进行更广泛的体内研究。对来自基底样PDX WHIM 2的乳腺肿瘤进行单细胞RNA测序，所述基底样PDX WHIM 2用媒介物或厄洛替尼处理9周。然后在体内对PDX进行长期厄洛替尼治疗。通过连续传代，产生了WHIM 2的厄洛替尼抗性亚系。对亲代和厄洛替尼耐药肿瘤进行了批量RNA测序。在亲本和厄洛替尼抗性细胞上进行具有> 500种临床使用的化合物的体外高通量药物筛选。将先前发表的来自MMTV-Wnt1肿瘤的批量基因表达微阵列数据与WHIM2 PDX数据进行对比。厄洛替尼有效抑制WHIM2肿瘤生长约4周。与未处理的细胞相比，单细胞RNA测序显示厄洛替尼处理的细胞中更大比例处于细胞周期的G1期。WHIM2和MMTV-Wnt1基因表达数据的比较揭示了一组38个重叠基因在厄洛替尼耐药的WHIM2和MMTV-Wnt1肿瘤中差异表达。所有三种数据类型的比较揭示了在所有厄洛替尼耐药样本中上调的五种基因：IL 19、KLK 7、LCN 2、SAA 1和SAA 2。在这五个基因中，LCN 2在三阴性乳腺癌中表达最丰富，并且其敲低恢复了体外厄洛替尼的敏感性。尽管存在转录组学差异，但亲本和厄洛替尼耐药WHIM2对大多数体外细胞毒性评估药物的反应相似。这项研究确定了厄洛替尼耐药基底细胞样乳腺癌中出现的转录组学变化。这些数据可用于鉴定生物标志物或开发预测患者对EGFRi反应的基因签名。未来的研究应该探索这些基因特征的预测能力，以及LCN 2如何促进EGFR i耐药性的发展。
The goals of this study were to identify transcriptomic changes that arise in basal-like breast cancer cells during the development of resistance to epidermal growth factor receptor inhibitors (EGFRi) and to identify drugs that are cytotoxic once EGFRi resistance occurs. Human patient-derived xenografts (PDXs) were grown in immunodeficient mice and treated with a set of EGFRi; the EGFRi erlotinib was selected for more expansive in vivo studies. Single-cell RNA sequencing was performed on mammary tumors from the basal-like PDX WHIM2 that was treated with vehicle or erlotinib for 9 weeks. The PDX was then subjected to long-term erlotinib treatment in vivo. Through serial passaging, an erlotinib-resistant subline of WHIM2 was generated. Bulk RNA-sequencing was performed on parental and erlotinib-resistant tumors. In vitro high-throughput drug screening with > 500 clinically used compounds was performed on parental and erlotinib-resistant cells. Previously published bulk gene expression microarray data from MMTV-Wnt1 tumors were contrasted with the WHIM2 PDX data. Erlotinib effectively inhibited WHIM2 tumor growth for approximately 4 weeks. Compared to untreated cells, single-cell RNA sequencing revealed that a greater proportion of erlotinib-treated cells were in the G1 phase of the cell cycle. Comparison of WHIM2 and MMTV-Wnt1 gene expression data revealed a set of 38 overlapping genes that were differentially expressed in the erlotinib-resistant WHIM2 and MMTV-Wnt1 tumors. Comparison of all three data types revealed five genes that were upregulated across all erlotinib-resistant samples: IL19, KLK7, LCN2, SAA1, and SAA2. Of these five genes, LCN2 was most abundantly expressed in triple-negative breast cancers, and its knockdown restored erlotinib sensitivity in vitro. Despite transcriptomic differences, parental and erlotinib-resistant WHIM2 displayed similar responses to the majority of drugs assessed for cytotoxicity in vitro. This study identified transcriptomic changes arising in erlotinib-resistant basal-like breast cancer. These data could be used to identify a biomarker or develop a gene signature predictive of patient response to EGFRi. Future studies should explore the predictive capacity of these gene signatures as well as how LCN2 contributes to the development of EGFRi resistance.
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DOI：
10.1158/1078-0432.ccr-13-0799
发表时间：
2013-10-01
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Masuda H;Baggerly KA;Wang Y;Zhang Y;Gonzalez-Angulo AM;Meric-Bernstam F;Valero V;Lehmann BD;Pietenpol JA;Hortobagyi GN;Symmans WF;Ueno NT
通讯作者：
Ueno NT
影响因子：
12.3
作者：
Pfefferle AD;Herschkowitz JI;Usary J;Harrell JC;Spike BT;Adams JR;Torres-Arzayus MI;Brown M;Egan SE;Wahl GM;Rosen JM;Perou CM
通讯作者：
Perou CM
影响因子：
7.4
作者：
Alzubi, Mohammad A.;Turner, Tia H.;Harrell, J. Chuck
通讯作者：
Harrell, J. Chuck
影响因子：
3
作者：
Masuda, Hiroko;Masuda, Norikazu;Tsujinaka, Toshimasa
通讯作者：
Tsujinaka, Toshimasa
影响因子：
4.3
作者：
Pfefferle, Adam D.;Darr, David B.;Perout, Charles M.
通讯作者：
Perout, Charles M.