Pub Date : 2021-07-01DOI: 10.1158/1538-7445.AM2021-1942
Colten M. McEwan, David G. Broadbent, Ashari R Kannangara, Daniel M. Poole, Bradley C. Naylor, J. C. Price, Jens C. Schmidt, J. Andersen
{"title":"Abstract 1942: Quantitative D2O proteomics reveals ATG9A-dependent autophagy substrates that are degraded independently of the LC3-lipidation machinery","authors":"Colten M. McEwan, David G. Broadbent, Ashari R Kannangara, Daniel M. Poole, Bradley C. Naylor, J. C. Price, Jens C. Schmidt, J. Andersen","doi":"10.1158/1538-7445.AM2021-1942","DOIUrl":"https://doi.org/10.1158/1538-7445.AM2021-1942","url":null,"abstract":"","PeriodicalId":18754,"journal":{"name":"Molecular and Cellular Biology / Genetics","volume":"31 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87901473","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-07-01DOI: 10.1158/1538-7445.AM2021-2335
K. Cargill, C. Stewart, E. Park, R. Cardnell, Youhong Fan, Qi Wang, L. Diao, W. Chan, P. Lorenzi, Jing Wang, L. Byers
{"title":"Abstract 2335: Targeting MYC-enhanced glycolysis in small cell lung cancer","authors":"K. Cargill, C. Stewart, E. Park, R. Cardnell, Youhong Fan, Qi Wang, L. Diao, W. Chan, P. Lorenzi, Jing Wang, L. Byers","doi":"10.1158/1538-7445.AM2021-2335","DOIUrl":"https://doi.org/10.1158/1538-7445.AM2021-2335","url":null,"abstract":"","PeriodicalId":18754,"journal":{"name":"Molecular and Cellular Biology / Genetics","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88223942","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-07-01DOI: 10.1158/1538-7445.AM2021-2225
Nakul M. Shah, H. S. Jang, J. H. Maeng, S. Tzeng, A. Wu, Changxu Fan, Noah L Basri, B. Katz, Daofeng Li, X. Xing, B. Evans, Ting Wang
Transposable elements (TEs) represent close to half of the genome and are generally disregarded in genomic studies due to their silencing in somatic tissues and difficulty in mapping to their repetitive sequences. Recent studies have revealed that epigenetic dysregulation in cancer can unlock the regulatory potential of transposable elements (TEs), and they can play an important role in cancer progression and oncogenesis. One important consequence of this phenomenon is the pervasive activation of TEs9 intrinsic promoters, which leads to generation of thousands of unique transcripts. Many of these transcripts splice into downstream genes and lead to the generation of TE-gene chimeric transcripts. These transcripts can alter the main reading frame of the original transcript to generate unique isoforms of the target gene or generate novel out-of-frame peptides that could be therapeutic targets. In this study, we analyzed the transcriptomes of 11,092 samples from 33 TCGA cancer types and 675 cancer cell lines to comprehensively profile all TE-gene fusion transcripts. Using somatic tissues from FANTOM5 and GTEx, we filtered these transcripts for tumor-specificity and discovered 2,642 tumor-specific TE-gene transcripts that promiscuously occur in nearly all TCGA tumor samples. Computational prediction of reading frames of these transcripts identified 1,202 candidates with the potential to generate tumor-specific TE-derived antigens (TS-TEAs). We further analyzed tumor mass spectrometry data from breast adenocarcinoma and ovarian cancer and confirmed that unique peptide sequences from TS-TEAs could be detected. In addition, we performed HLA-pulldown mass spectrometry and confirmed that TS-TEAs are presented on the cell surface in cancer cell lines. Given that these antigens are highly shared within and across cancer types, we assessed their potential to generate universal antigen-based therapies. Optimal combinations of 5, 10, and 20 TS-TEAs could generate unique peptides that bind to patient-specific HLA alleles for 39.2%, 50.8%, and 60.8% of all TCGA tumors respectively. Lastly, we highlight the tumor-specific membrane proteins transcribed from TE-exapted promoters that can potentially expose novel epitopes on the extracellular surface of cancer cells. These can be valuable targets of CAR-T or alternative antibody-based therapies. In conclusion, we showcase the high prevalence of TE-derived promoter activation in cancer and suggest multiple avenues by which this phenomenon can be targeted therapeutically. Citation Format: Nakul M. Shah, Hyo Sik Jang, Ju Heon Maeng, Shin-Cheng Tzeng, Angela Wu, Changxu Fan, Noah L. Basri, Benjamin Katz, Daofeng Li, Xiaoyun Xing, Bradley S. Evans, Ting Wang. Transposable elements are an abundant and pan-cancer source of shared tumor-specific antigens and membrane targets [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR;
转座因子(te)占基因组的近一半，由于其在体细胞组织中沉默且难以定位到其重复序列，因此在基因组研究中通常被忽视。最近的研究表明，癌症的表观遗传失调可以释放转座因子(te)的调控潜力，它们在癌症的进展和肿瘤发生中发挥重要作用。这一现象的一个重要后果是TEs9内在启动子的普遍激活，导致数千个独特转录本的产生。许多这些转录物拼接到下游基因中，导致te基因嵌合转录物的产生。这些转录物可以改变原始转录物的主要阅读框，以产生独特的靶基因同工型，或产生可能成为治疗靶点的新的框架外肽。在这项研究中，我们分析了来自33种TCGA癌症类型和675种癌细胞系的11092个样本的转录组，以全面分析所有te基因融合转录物。使用来自FANTOM5和GTEx的体细胞组织，我们过滤了这些转录本的肿瘤特异性，发现了2,642个肿瘤特异性te基因转录本，这些转录本几乎在所有TCGA肿瘤样本中都混杂存在。对这些转录本的阅读框进行计算预测，确定了1202个候选基因，它们有可能产生肿瘤特异性te衍生抗原(TS-TEAs)。我们进一步分析了乳腺腺癌和卵巢癌的肿瘤质谱数据，证实了ts - tea的独特肽序列可以被检测到。此外，我们进行了hla -pull质谱分析，证实了ts - tea在癌细胞系的细胞表面存在。鉴于这些抗原在癌症类型内部和不同类型之间高度共享，我们评估了它们产生通用抗原基础疗法的潜力。5、10和20个ts - tea的最佳组合可以产生独特的肽，分别与39.2%、50.8%和60.8%的TCGA肿瘤患者特异性HLA等位基因结合。最后，我们强调了从te外显子转录的肿瘤特异性膜蛋白，这些膜蛋白可能会在癌细胞的细胞外表面暴露新的表位。这些可能是CAR-T或其他基于抗体的疗法的有价值的靶点。总之，我们展示了te衍生的启动子激活在癌症中的高患病率，并提出了多种途径，通过这种现象可以靶向治疗。引用格式:Nakul M. Shah, Hyo Sik Jang, Ju Heon Maeng，曾信成，Angela Wu，樊昌旭，Noah L. Basri, Benjamin Katz，李道峰，邢晓云，Bradley S. Evans，王婷转座因子是共享肿瘤特异性抗原和膜靶点的丰富和泛癌来源[摘要]。见:美国癌症研究协会2021年年会论文集;2021年4月10日至15日和5月17日至21日。费城(PA): AACR;癌症杂志，2021;81(13 -增刊):2225。
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Pub Date : 2021-07-01DOI: 10.1158/1538-7445.AM2021-86
Lorela Ciraku, Zachary A. Bacigalupa, Rebecca A. Moeller, Jing Ju, Rusia Lee, C. Ferrer, S. Trefely, N. Snyder, L. D'agostino, C. Katsetos, Wenyin Shi, M. Reginato
Cancer cells alter their metabolism to increase cell growth. A subset of the glucose taken up is shunted into the hexosamine biosynthetic pathway where it is used to synthesize UDP-GlcNAc, a substrate of O-GlcNAc transferase (OGT), which modifies cytoplasmic and nuclear proteins with O-linked sugar moieties. Here, we show that OGT and O-GlcNAcylation are elevated in glioblastoma (GBM) cancer cells and in GBM patient samples that correlates with disease progression. Reduction of OGT expression in GBM cells led to significant reduction in anchorage-independent growth, acetyl-CoA levels, and decrease in free fatty acids. Conversely, overexpressing OGT in GBM cells had the opposite effect. Reducing OGT expression in GBMcells transplanted in an orthotopic intracranial mouse model reduced tumor growth and extended survival. Mechanistically, we show that OGT overexpression increases carbon-flux of acetate to acetyl-CoA, a reaction carried by the enzyme acetyl-CoA synthetase 2 (ACSS2). Indeed, OGT regulates ACSS2 protein levels and O-GlcNAcylation increases ACSS2phosphorylation on Ser-267 in a cyclin dependent kinase 5 (CDK5)-dependent manner, which regulates its stability by reducing polyubiquitination and degradation. ACSS2 Ser-267 is critical for OGT-mediated GBM growth as overexpression of ACSS2 Ser-267 phospho-mimetic rescues growth in vitro and in vivo. Using an ex vivo GBM brain slice model we show that treatment of GBM-transplanted slices with OGT inhibitor Ac-GlcNAc-5S or pan-cdk inhibitor dinaciclib reduced growth of pre-formed tumors in cultured brain slices. These results suggest a crucial role for O-GlcNAc signaling in transducing nutritional state to regulate acetate metabolism and identify OGT and CDK5 as novel therapeutic targets for treatment of glioblastoma. Citation Format: Lorela Ciraku, Zachary Bacigalupa, Rebecca Moeller, Jing Ju, Rusia H. Lee, Christina Ferrer, Sophie Trefely, Nathaniel W. Snyder, Luca D'Agostino, Christos D. Katsetos, Wenyin Shi, Mauricio J. Reginato. O-GlcNAc transferase regulates glioblastoma acetate metabolism via regulation of CDK5-dependent ACSS2 phosphorylation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 86.
癌细胞通过改变新陈代谢来促进细胞生长。摄取的一部分葡萄糖被分流到己糖胺生物合成途径中，在那里它被用来合成UDP-GlcNAc，一种O-GlcNAc转移酶(OGT)的底物，它用o -连接糖部分修饰细胞质和核蛋白。在这里，我们发现OGT和o - glcn酰化在胶质母细胞瘤(GBM)癌细胞和GBM患者样本中升高，这与疾病进展有关。GBM细胞中OGT表达的降低导致锚定非依赖性生长、乙酰辅酶a水平和游离脂肪酸的显著降低。相反，在GBM细胞中过表达OGT具有相反的效果。在原位颅内小鼠模型移植的gbcells中，降低OGT表达可减少肿瘤生长并延长生存期。从机制上讲，我们发现OGT过表达增加了醋酸酯到乙酰辅酶a的碳通量，这是一个由乙酰辅酶a合成酶2 (ACSS2)进行的反应。事实上，OGT调节ACSS2蛋白水平，o - glcn酰化以细胞周期蛋白依赖激酶5 (CDK5)依赖的方式增加Ser-267上的ACSS2磷酸化，从而通过减少多泛素化和降解来调节其稳定性。ACSS2 Ser-267对ogt介导的GBM生长至关重要，因为ACSS2 Ser-267的过表达可以挽救体外和体内的生长。通过离体GBM脑切片模型，我们发现用OGT抑制剂Ac-GlcNAc-5S或pan-cdk抑制剂dinaciclib处理GBM移植的脑切片可以减少培养脑切片中预形成肿瘤的生长。这些结果提示O-GlcNAc信号在营养状态的传导中发挥重要作用，调节醋酸盐代谢，并确定OGT和CDK5作为治疗胶质母细胞瘤的新靶点。引文格式:Lorela Ciraku, Zachary Bacigalupa, Rebecca Moeller, Jing Ju, russia H. Lee, Christina Ferrer, Sophie Trefely, Nathaniel W. Snyder, Luca D'Agostino, Christos D. Katsetos, Wenyin Shi, Mauricio J. Reginato。O-GlcNAc转移酶通过调节cdk5依赖性ACSS2磷酸化来调节胶质母细胞瘤醋酸盐代谢[摘要]。见:美国癌症研究协会2021年年会论文集;2021年4月10日至15日和5月17日至21日。费城(PA): AACR;癌症杂志，2021;81(13 -增刊):摘要第86期。
{"title":"Abstract 86: O-GlcNAc transferase regulates glioblastoma acetate metabolism via regulation of CDK5-dependent ACSS2 phosphorylation","authors":"Lorela Ciraku, Zachary A. Bacigalupa, Rebecca A. Moeller, Jing Ju, Rusia Lee, C. Ferrer, S. Trefely, N. Snyder, L. D'agostino, C. Katsetos, Wenyin Shi, M. Reginato","doi":"10.1158/1538-7445.AM2021-86","DOIUrl":"https://doi.org/10.1158/1538-7445.AM2021-86","url":null,"abstract":"Cancer cells alter their metabolism to increase cell growth. A subset of the glucose taken up is shunted into the hexosamine biosynthetic pathway where it is used to synthesize UDP-GlcNAc, a substrate of O-GlcNAc transferase (OGT), which modifies cytoplasmic and nuclear proteins with O-linked sugar moieties. Here, we show that OGT and O-GlcNAcylation are elevated in glioblastoma (GBM) cancer cells and in GBM patient samples that correlates with disease progression. Reduction of OGT expression in GBM cells led to significant reduction in anchorage-independent growth, acetyl-CoA levels, and decrease in free fatty acids. Conversely, overexpressing OGT in GBM cells had the opposite effect. Reducing OGT expression in GBMcells transplanted in an orthotopic intracranial mouse model reduced tumor growth and extended survival. Mechanistically, we show that OGT overexpression increases carbon-flux of acetate to acetyl-CoA, a reaction carried by the enzyme acetyl-CoA synthetase 2 (ACSS2). Indeed, OGT regulates ACSS2 protein levels and O-GlcNAcylation increases ACSS2phosphorylation on Ser-267 in a cyclin dependent kinase 5 (CDK5)-dependent manner, which regulates its stability by reducing polyubiquitination and degradation. ACSS2 Ser-267 is critical for OGT-mediated GBM growth as overexpression of ACSS2 Ser-267 phospho-mimetic rescues growth in vitro and in vivo. Using an ex vivo GBM brain slice model we show that treatment of GBM-transplanted slices with OGT inhibitor Ac-GlcNAc-5S or pan-cdk inhibitor dinaciclib reduced growth of pre-formed tumors in cultured brain slices. These results suggest a crucial role for O-GlcNAc signaling in transducing nutritional state to regulate acetate metabolism and identify OGT and CDK5 as novel therapeutic targets for treatment of glioblastoma. Citation Format: Lorela Ciraku, Zachary Bacigalupa, Rebecca Moeller, Jing Ju, Rusia H. Lee, Christina Ferrer, Sophie Trefely, Nathaniel W. Snyder, Luca D'Agostino, Christos D. Katsetos, Wenyin Shi, Mauricio J. Reginato. O-GlcNAc transferase regulates glioblastoma acetate metabolism via regulation of CDK5-dependent ACSS2 phosphorylation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 86.","PeriodicalId":18754,"journal":{"name":"Molecular and Cellular Biology / Genetics","volume":"23 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82817227","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-07-01DOI: 10.1158/1538-7445.AM2021-2001
Y. Tsai, Su-Liang Chen, S. Peng, Kuang-Hung Cheng, S. Jiang, S. Chuang, Chang Hui-ju
{"title":"Abstract 2001: Krupple like factor 10 modulates stem cell phenotype of pancreatic adenocarcinoma via transcriptional regulation of Notch signal pathway","authors":"Y. Tsai, Su-Liang Chen, S. Peng, Kuang-Hung Cheng, S. Jiang, S. Chuang, Chang Hui-ju","doi":"10.1158/1538-7445.AM2021-2001","DOIUrl":"https://doi.org/10.1158/1538-7445.AM2021-2001","url":null,"abstract":"","PeriodicalId":18754,"journal":{"name":"Molecular and Cellular Biology / Genetics","volume":"57 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86750700","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-07-01DOI: 10.1158/1538-7445.AM2021-2240
Shankara K. Anand, M. Bustoros, F. Aguet, R. Sklavenitis-Pistofidis, R. Redd, Binyamin Zhitomirsky, Andrew J. Dunford, Y. Tai, S. Chavda, Cody J Boehner, C. Neuse, T. Casneuf, L. Trippa, C. Stewart, K. Yong, I. Ghobrial, G. Getz
{"title":"Abstract 2240: Genomic profiling of smoldering multiple myeloma classifies distinct molecular groups","authors":"Shankara K. Anand, M. Bustoros, F. Aguet, R. Sklavenitis-Pistofidis, R. Redd, Binyamin Zhitomirsky, Andrew J. Dunford, Y. Tai, S. Chavda, Cody J Boehner, C. Neuse, T. Casneuf, L. Trippa, C. Stewart, K. Yong, I. Ghobrial, G. Getz","doi":"10.1158/1538-7445.AM2021-2240","DOIUrl":"https://doi.org/10.1158/1538-7445.AM2021-2240","url":null,"abstract":"","PeriodicalId":18754,"journal":{"name":"Molecular and Cellular Biology / Genetics","volume":"85 2","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"91463228","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-07-01DOI: 10.1158/1538-7445.AM2021-2042
Menuka Karki, Rahul K. Jangid, R. Anish, Riyad N. H. Seervai, Jean-Philippe Bertocchio, Takashi Hotta, P. Msaouel, S. Jung, S. Grimm, C. Coarfa, B. Weissman, R. Ohi, K. Verhey, Courtney Hodges, R. Dere, I. Park, B. Prasad, W. K. Rathmell, Cheryl Walker, D. Tripathi
{"title":"Abstract 2042: A cytoskeletal function for PBRM1: reading methylated microtubules to maintain genomic stability","authors":"Menuka Karki, Rahul K. Jangid, R. Anish, Riyad N. H. Seervai, Jean-Philippe Bertocchio, Takashi Hotta, P. Msaouel, S. Jung, S. Grimm, C. Coarfa, B. Weissman, R. Ohi, K. Verhey, Courtney Hodges, R. Dere, I. Park, B. Prasad, W. K. Rathmell, Cheryl Walker, D. Tripathi","doi":"10.1158/1538-7445.AM2021-2042","DOIUrl":"https://doi.org/10.1158/1538-7445.AM2021-2042","url":null,"abstract":"","PeriodicalId":18754,"journal":{"name":"Molecular and Cellular Biology / Genetics","volume":"11 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2021-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88939480","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2021-07-01DOI: 10.1158/1538-7445.AM2021-LB186
Rebecca Caeser, Christopher H. Hulton, Vidushi Durani, Emily A. Costa, M. Little, Nisargbhai S. Shah, E. Stanchina, J. Poirier, C. Rudin, T. Sen
Background Lung cancer is the leading cause of cancer death, killing more people than colon, breast, and prostate cancers combined (Siegel et al. 2013). Small cell lung cancer (SCLC) is a high grade neuroendocrine tumor accounting for ~15% of all lung cancers (Hann et al. 2019). Metastasis is often found at first diagnosis, making SCLC exceptionally lethal (2-year survival l Gao et al. 2013; Wagle et al. 2018). This striking difference is not well understood and previous attempts to determine whether this might be therapeutically important (Ravi et al, 1998; Cristea et al. 2020) have had conflicting conclusions. SCLC has recently been defined by the relative expression of four major transcriptional regulators (ASCL1, NeuroD1, POU2F3, YAP1) (Rudin et al., 2019). In this study we aimed to elucidate the effect of MAPK activation in these different SCLC subtypes and explore its therapeutic vulnerability. Results We used a doxycycline-inducible vector for expression of MEKDDS217D/S221D (MEK1) in a cohort of ASCL1-, NEUROD1, and POU2F3- driven cell lines. Activation through MEK1 in ASCL1-driven SCLC cell lines resulted in a significant decrease in cell growth over 9 days. This was associated with a decrease in neuroendocrine markers ASCL1 and INSM1, a G2 cell cycle arrest and no significant increase in apoptotic cells. Expression of MEK1 in other SCLC subtypes and NSCLC failed to show any appreciable changes in cell growth. Remarkably, athymic mice injected with a MEK1 expressing ASCL1-driven cell line showed significantly slower tumor formation and longer survival than the ASCL1-driven cell line not expressing MEK1. Previous work established that hyperactivation of BRAFV600E, RAS and MYC can result in oncogene-induced senescence (Serrano et al. 1997) which is caused by upregulation of negative feedback such as SPRY2, DUSP6, ETV5 rather than MAPK pathway activation in some solid tumors and pre-B ALL (Courtois-Cox et al 2006; Shojaee et al. 2015). Similarly, we also observed strong upregulation of DUSP6, SPRY2, but not ETV5 upon MAPK activation. This was especially prominent in ASCL1-driven cell lines that changed from the normal phenotype of being in suspension to a more adherent morphology as a result of MAPK activation. Interestingly, phosphokinase array in the major subtype cell lines after MEK1 activation, demonstrated that, almost exclusively, the STAT pathways, in particular STAT3 through phosphorylation at S727 was strongly upregulated in the ASCL1-driven subtype. This prompted us to examine whether these cells were sensitive to STAT3 inhibition. Upon treatment with a STAT3 inhibitor, Stattic (1μM), ASCL1-driven SCLC cells reached their IC50 after 3-5 days in comparison to 9 days for other SCLC subtypes. NSCLC cells were resistant to STAT3 inhibition. Summary These findings suggest that ASCL1-driven SCLC in vitro and in vivo is sensitive to activation of MAPK signaling in comparison to other SCLC subtypes. Whilst activation of the MAPK pathway m
肺癌是癌症死亡的主要原因，死亡人数超过结肠癌、乳腺癌和前列腺癌的总和(Siegel et al. 2013)。小细胞肺癌(Small cell lung cancer, SCLC)是一种高度神经内分泌肿瘤，约占所有肺癌的15% (Hann et al. 2019)。转移通常在首次诊断时就发现，这使得SCLC异常致命(2年生存率)。Wagle et al. 2018)。这种显著的差异并没有被很好地理解，之前的尝试确定这是否可能在治疗上很重要(Ravi等人，1998;Cristea et al. 2020)得出了相互矛盾的结论。SCLC最近被定义为四种主要转录调控因子(ASCL1、NeuroD1、POU2F3、YAP1)的相对表达(Rudin et al.， 2019)。在这项研究中，我们旨在阐明MAPK激活在这些不同SCLC亚型中的作用，并探讨其治疗易感性。我们使用强力霉素诱导的载体在ASCL1-、NEUROD1和POU2F3驱动的细胞系中表达MEKDDS217D/S221D (MEK1)。在ascl1驱动的SCLC细胞系中，通过MEK1激活导致细胞生长在9天内显著下降。这与神经内分泌标志物ASCL1和INSM1的减少、G2细胞周期阻滞和凋亡细胞无显著增加有关。MEK1在其他SCLC亚型和NSCLC中的表达在细胞生长过程中没有明显变化。值得注意的是，与不表达MEK1的ascl1驱动细胞系相比，注射表达MEK1的ascl1驱动细胞系的胸腺小鼠肿瘤形成速度明显减慢，存活时间更长。先前的研究证实，BRAFV600E、RAS和MYC的过度激活可导致癌基因诱导的衰老(Serrano et al. 1997)，这是由一些实体肿瘤和b前ALL中SPRY2、DUSP6、ETV5等负反馈的上调而不是MAPK通路的激活引起的(Courtois-Cox et al. 2006;Shojaee et al. 2015)。同样，我们也观察到在MAPK激活后DUSP6、SPRY2明显上调，但ETV5不上调。这在ascl1驱动的细胞系中尤其突出，由于MAPK激活，这些细胞系从悬浮的正常表型转变为更粘附的形态。有趣的是，在MEK1激活后，主要亚型细胞系中的磷酸激酶阵列表明，几乎完全是STAT途径，特别是通过S727磷酸化的STAT3在ascl1驱动的亚型中被强烈上调。这促使我们研究这些细胞是否对STAT3抑制敏感。用STAT3抑制剂Stattic (1μM)治疗后，ascl1驱动的SCLC细胞在3-5天后达到IC50，而其他SCLC亚型则为9天。NSCLC细胞对STAT3抑制具有抗性。这些研究结果表明，与其他SCLC亚型相比，ascl1驱动的SCLC在体外和体内对MAPK信号的激活敏感。虽然激活MAPK通路似乎与当前旨在抑制致癌信号的治疗策略相违背，但我们建议使用一种STAT3抑制剂，该抑制剂在体外已被证明是有效的。引用形式:Rebecca Caeser, Christopher Hulton, Vidushi Durani, Emily Costa, Megan Little, Nisargbhai S. Shah, Elisa de Stanchina, John T. Poirier, Charles M. Rudin, Triparna Sen. MAPK通路激活代表ascl1驱动的SCLC的治疗脆弱性[摘要]。见:美国癌症研究协会2021年年会论文集;2021年4月10日至15日和5月17日至21日。费城(PA): AACR;癌症杂志，2021;81(13 -增刊):摘要nr LB186。
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