Pub Date : 2019-02-01DOI: 10.1158/2326-6074.CRICIMTEATIAACR18-B163
Chun-Chieh Hsu, Maudry Laurent-Rolle, P. Cresswell
The innate immune response involves the induction of hundreds of interferon-stimulated genes (ISGs), many of which play a role in cancer immunosurveillance and antiviral immunity. Therefore, understanding how ISGs function and coordinate with the cellular network is of critical importance. We focus on one such ISG, viperin (virus inhibitory protein, endoplasmic reticulum-associated, interferon-inducible). Viperin has been reported to inhibit a broad spectrum of DNA and RNA viruses, including many flaviviruses, human cytomegalovirus (HCMV), Chikungunya virus, influenza A virus, Sindbis virus, vesicular stomatitis virus, tick-borne encephalitis virus, HIV-1 and more. It is highly conserved in evolution, from protozoans to humans. Viperin is a peripheral membrane protein, associating on the cytosolic face of the ER via its N-terminal amphipathic helix. The cytosolic domain contains a radical SAM domain with a [4Fe-4S] cluster coordinated by three cysteine residues in the active site. Although the enzymatic function and substrate of viperin is still under debate, a recent study reveals that viperin catalyzes the conversion of cytidine triphosphate to 3′-deoxy-3′,4′-didehydro-CTP (ddhCTP) via its radical SAM activity. However, a general mechanism for viperin action and its potential roles within the innate immune response remain to be defined. Here, we demonstrate that viperin inhibits global protein translation via its radical SAM-dependent enzymatic activity. Using immortalized mouse macrophages from wild-type and viperin knockout mice we show that, although other ISGs are induced, type I IFN treatment of WT but not viperin KO mouse microphages leads to a global change in polysome profile with an increase in the ribosome fraction and a decrease in actively translating ribosomes. In parallel, the translational regulation function was evaluated in 293T-cells with transient transfection and doxycycline-inducible system. Despite no IFN stimulation and therefore no other ISG expression, viperin effectively represses translation. Furthermore, we find that viperin reduces global protein translation by ~30% in live cells using [35S]-methionine metabolic pulse-labeling or O-propargyl-puromycin (OPP) labelling. The radical SAM enzymatic activity is required for translational inhibition: site-directed mutagenesis of amino residues involved in iron-sulfur cluster and substrate binding abolishes the translational inhibition activity. We also demonstrate that ddhCTP, a recently identified vipern product, inhibits global protein translation in live cells. Viral replication requires the host translation machinery to make viral proteins, and our results suggest that much of its antiviral activity may be attributable to translational regulation. We show that viperin inhibits viral protein synthesis and viral replication of the Kunjin virus, a West Nile Virus variant, in tissue culture cells. Our study suggests a partially unifying mechanism for the broad antiviral fu
先天性免疫反应涉及诱导数百个干扰素刺激基因（ISGs），其中许多基因在癌症免疫监视和抗病毒免疫中发挥作用。因此，了解 ISGs 如何发挥作用并与细胞网络协调至关重要。我们重点研究了这样一种 ISG：viperin（病毒抑制蛋白，内质网相关，干扰素诱导）。据报道，蝰蛇蛋白可抑制多种 DNA 和 RNA 病毒，包括多种黄病毒、人类巨细胞病毒（HCMV）、基孔肯雅病毒、甲型流感病毒、辛比斯病毒、水泡性口炎病毒、蜱传脑炎病毒、HIV-1 等。从原生动物到人类，它在进化过程中高度保守。蝰蛇素是一种外周膜蛋白，通过其 N 端两性螺旋与 ER 的胞浆面结合。细胞膜结构域包含一个自由基 SAM 结构域，其中的[4Fe-4S]簇与活性位点中的三个半胱氨酸残基相协调。尽管蝰蛇素的酶功能和底物仍存在争议，但最近的一项研究发现，蝰蛇素通过其自由基 SAM 活性催化三磷酸胞苷转化为 3′-脱氧-3′,4′-二脱氢-CTP（ddhCTP）。然而，毒蛇素作用的一般机制及其在先天性免疫反应中的潜在作用仍有待明确。在这里，我们证明毒蛇素通过其自由基 SAM 依赖性酶活性抑制全局蛋白质翻译。通过使用野生型和毒蛇素基因敲除小鼠的永生化小鼠巨噬细胞，我们发现，尽管诱导了其他 ISGs，但对 WT 而非毒蛇素基因敲除小鼠巨噬细胞进行 I 型 IFN 处理会导致多聚体轮廓的整体变化，即核糖体部分增加，活跃翻译的核糖体减少。与此同时，通过瞬时转染和强力霉素诱导系统对 293T 细胞的翻译调节功能进行了评估。尽管没有 IFN 的刺激，因此也没有其他 ISG 的表达，毒蛇素仍有效地抑制了翻译。此外，我们发现毒蛇素在活细胞中使用[35S]-蛋氨酸代谢脉冲标记或 O-丙炔-尿霉素（OPP）标记可使全局蛋白质翻译减少约 30%。翻译抑制需要自由基 SAM 酶活性：对参与铁硫簇和底物结合的氨基酸残基进行定点突变后，翻译抑制活性消失。我们还证明，最近发现的一种毒蛋白产物 ddhCTP 可抑制活细胞中的全局蛋白质翻译。病毒复制需要宿主翻译机制来制造病毒蛋白，我们的研究结果表明，毒蛇素的大部分抗病毒活性可能归因于翻译调控。我们的研究表明，毒蛇素能抑制昆金病毒（一种西尼罗河病毒变种）在组织培养细胞中的病毒蛋白合成和病毒复制。我们的研究提示了基于翻译调节的毒蛇素广泛抗病毒功能的部分统一机制。引用格式：徐春杰，莫德里-劳伦特-罗尔，彼得-克莱斯威尔。干扰素诱导的抗病毒蛋白毒蛇素对翻译的调控[摘要].In：第四届CRI-CIMT-EATI-AACR国际癌症免疫疗法会议论文集：将科学转化为生存；2018年9月30日-10月3日；纽约州纽约市。费城（宾夕法尼亚州）：AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr B163.
{"title":"Abstract B163: Regulation of translation by the interferon-induced antiviral protein viperin","authors":"Chun-Chieh Hsu, Maudry Laurent-Rolle, P. Cresswell","doi":"10.1158/2326-6074.CRICIMTEATIAACR18-B163","DOIUrl":"https://doi.org/10.1158/2326-6074.CRICIMTEATIAACR18-B163","url":null,"abstract":"The innate immune response involves the induction of hundreds of interferon-stimulated genes (ISGs), many of which play a role in cancer immunosurveillance and antiviral immunity. Therefore, understanding how ISGs function and coordinate with the cellular network is of critical importance. We focus on one such ISG, viperin (virus inhibitory protein, endoplasmic reticulum-associated, interferon-inducible). Viperin has been reported to inhibit a broad spectrum of DNA and RNA viruses, including many flaviviruses, human cytomegalovirus (HCMV), Chikungunya virus, influenza A virus, Sindbis virus, vesicular stomatitis virus, tick-borne encephalitis virus, HIV-1 and more. It is highly conserved in evolution, from protozoans to humans. Viperin is a peripheral membrane protein, associating on the cytosolic face of the ER via its N-terminal amphipathic helix. The cytosolic domain contains a radical SAM domain with a [4Fe-4S] cluster coordinated by three cysteine residues in the active site. Although the enzymatic function and substrate of viperin is still under debate, a recent study reveals that viperin catalyzes the conversion of cytidine triphosphate to 3′-deoxy-3′,4′-didehydro-CTP (ddhCTP) via its radical SAM activity. However, a general mechanism for viperin action and its potential roles within the innate immune response remain to be defined. Here, we demonstrate that viperin inhibits global protein translation via its radical SAM-dependent enzymatic activity. Using immortalized mouse macrophages from wild-type and viperin knockout mice we show that, although other ISGs are induced, type I IFN treatment of WT but not viperin KO mouse microphages leads to a global change in polysome profile with an increase in the ribosome fraction and a decrease in actively translating ribosomes. In parallel, the translational regulation function was evaluated in 293T-cells with transient transfection and doxycycline-inducible system. Despite no IFN stimulation and therefore no other ISG expression, viperin effectively represses translation. Furthermore, we find that viperin reduces global protein translation by ~30% in live cells using [35S]-methionine metabolic pulse-labeling or O-propargyl-puromycin (OPP) labelling. The radical SAM enzymatic activity is required for translational inhibition: site-directed mutagenesis of amino residues involved in iron-sulfur cluster and substrate binding abolishes the translational inhibition activity. We also demonstrate that ddhCTP, a recently identified vipern product, inhibits global protein translation in live cells. Viral replication requires the host translation machinery to make viral proteins, and our results suggest that much of its antiviral activity may be attributable to translational regulation. We show that viperin inhibits viral protein synthesis and viral replication of the Kunjin virus, a West Nile Virus variant, in tissue culture cells. Our study suggests a partially unifying mechanism for the broad antiviral fu","PeriodicalId":120683,"journal":{"name":"Other Topics","volume":"70 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2019-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"126798677","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 : 2019-02-01DOI: 10.1158/2326-6074.CRICIMTEATIAACR18-B170
Julie Leca
Acute myeloid leukemia (AML) and angioimmunoblastic T-cell lymphoma (AITL), are hematologic diseases requiring novel approaches to patient selection and therapy. In AITL, neoplastic cells express many markers of T follicular helper (TFH) cells, while in AML, myeloid stem and progenitor cells are affected. Tumor cells of AML and AITL patients frequently bear mutations affecting genes involved in epigenetic regulation, including isocitrate dehydrogenase (IDH) and ten-eleven translocation-2 (TET2). IDH mutations drive production of the rare metabolite D-2-hydroxyglutarate (2HG), which competitively inhibits α-ketoglutarate (α-KG)-dependent dioxygenases such as the TET proteins (affecting DNA methylation) and Jumonji histone demethylases (altering histone methylation). IDH mutations occur in 30% of AML and AITL cases but the spectrum of these mutations differs. In AML, IDH1R132 (41%), IDH2R140 (44%) and IDH2R172 (15%) dominate, and IDH and TET2 mutations are mutually exclusive. In AITL, IDH2R140 (2%) and IDH2R172 (98%) dominate (no IDH1 mutations), with co-mutation of TET2 in 82.1% of cases. In AML, loss of these dioxygenase activities causes epigenetic alterations to DNA and histones, leads to abnormal gene transcription that affects hematopoietic cell differentiation, and drives myeloid disease. However, in AITL disease, the impact of the IDH2 mutation is completely unknown. My goal is to dissect the effects of IDH2 and TET2 mutations in hematologic diseases to understand how IDH2 and TET2 mutations collaborate to drive malignancy in AITL, and why they cooperate differently in myeloid and lymphoid diseases. I focus on DDR signaling and epigenetics analysis to found novel therapeutic vulnerabilities arise from the altered epigenetic regulation and DDR linked to IDH2 and TET2 mutations. I used a CD4-Cre mouse model to introduce the IDH2R172K and TET2 mutations in the T-cell compartment. This would allow me to study the collaboration between IDH2 and TET2 mutations in the context of T-cells. Mice bearing both IDH2 and TET2 mutations show decreased survival, with a median survival of approximately 8 months accompanied by splenomegaly and lymphadenopathy. This is significantly different than CD4+ control mice or single mutant mice. Double mutant mice (DM) present a disruption of spleen and lymph node architecture. To understand this phenotype, I performed comprehensive flow cytometry staining and, surprisingly, I found that the increased spleen size is not due to T-cell infiltration, but is due to increased erythropoiesis and expansion of immature erythropoietic cells (CD71+, cKit+). We can explain this result by the fact that CD4-Cre is also expressed in some progenitor cells leading to stress induced erythropoiesis. However, if we focus on the T-cell population, we see that DM mice present a T-cell phenotype including a decrease of CD4+ naive cells and an increase of CD4+ effector memory cells as early as 5 months. So, there is an imbalance in T-cell
{"title":"Abstract B170: Therapeutic implications of altered epigenetics and DNA damage responses in IDH2-mutated hematologic diseases","authors":"Julie Leca","doi":"10.1158/2326-6074.CRICIMTEATIAACR18-B170","DOIUrl":"https://doi.org/10.1158/2326-6074.CRICIMTEATIAACR18-B170","url":null,"abstract":"Acute myeloid leukemia (AML) and angioimmunoblastic T-cell lymphoma (AITL), are hematologic diseases requiring novel approaches to patient selection and therapy. In AITL, neoplastic cells express many markers of T follicular helper (TFH) cells, while in AML, myeloid stem and progenitor cells are affected. Tumor cells of AML and AITL patients frequently bear mutations affecting genes involved in epigenetic regulation, including isocitrate dehydrogenase (IDH) and ten-eleven translocation-2 (TET2). IDH mutations drive production of the rare metabolite D-2-hydroxyglutarate (2HG), which competitively inhibits α-ketoglutarate (α-KG)-dependent dioxygenases such as the TET proteins (affecting DNA methylation) and Jumonji histone demethylases (altering histone methylation). IDH mutations occur in 30% of AML and AITL cases but the spectrum of these mutations differs. In AML, IDH1R132 (41%), IDH2R140 (44%) and IDH2R172 (15%) dominate, and IDH and TET2 mutations are mutually exclusive. In AITL, IDH2R140 (2%) and IDH2R172 (98%) dominate (no IDH1 mutations), with co-mutation of TET2 in 82.1% of cases. In AML, loss of these dioxygenase activities causes epigenetic alterations to DNA and histones, leads to abnormal gene transcription that affects hematopoietic cell differentiation, and drives myeloid disease. However, in AITL disease, the impact of the IDH2 mutation is completely unknown. My goal is to dissect the effects of IDH2 and TET2 mutations in hematologic diseases to understand how IDH2 and TET2 mutations collaborate to drive malignancy in AITL, and why they cooperate differently in myeloid and lymphoid diseases. I focus on DDR signaling and epigenetics analysis to found novel therapeutic vulnerabilities arise from the altered epigenetic regulation and DDR linked to IDH2 and TET2 mutations. I used a CD4-Cre mouse model to introduce the IDH2R172K and TET2 mutations in the T-cell compartment. This would allow me to study the collaboration between IDH2 and TET2 mutations in the context of T-cells. Mice bearing both IDH2 and TET2 mutations show decreased survival, with a median survival of approximately 8 months accompanied by splenomegaly and lymphadenopathy. This is significantly different than CD4+ control mice or single mutant mice. Double mutant mice (DM) present a disruption of spleen and lymph node architecture. To understand this phenotype, I performed comprehensive flow cytometry staining and, surprisingly, I found that the increased spleen size is not due to T-cell infiltration, but is due to increased erythropoiesis and expansion of immature erythropoietic cells (CD71+, cKit+). We can explain this result by the fact that CD4-Cre is also expressed in some progenitor cells leading to stress induced erythropoiesis. However, if we focus on the T-cell population, we see that DM mice present a T-cell phenotype including a decrease of CD4+ naive cells and an increase of CD4+ effector memory cells as early as 5 months. So, there is an imbalance in T-cell ","PeriodicalId":120683,"journal":{"name":"Other Topics","volume":"73 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2019-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"126382019","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 : 2019-02-01DOI: 10.1158/2326-6074.CRICIMTEATIAACR18-B147
K. Blažková, Petr Šimon, Tomáš Knedlík, P. Dvořáková, A. Březinová, L. Kostka, V. Šubr, J. Konvalinka, P. Šácha
The renaissance of phenotypic screening leads to the discovery of biologically active small molecules resulting in a high need to quickly and efficiently identify their protein target. This is most commonly done by a preparation of an affinity probe for the target pull-down, which requires time-consuming structure-activity relationship studies to identify proper position for an attachment of an affinity anchor with necessary linker. We set out to circumvent this step altogether and prepare stochastic conjugates that would enable fast target identification without prior structural information or testing of suitable linker attachment position. We suggest that they could be used also for the identification of unknown targets of phenotypically active compounds as well as off-targets of established active compounds. To test the new approach, we selected several model protein targets – glutamate carboxypeptidase II (GCPII, also known as prostate specific membrane antigen, PSMA) expressed in prostate cancer, fibroblast activation protein (FAP) in tumor stroma, and aspartic protease family, specifically pepsin, cathepsin D and HIV protease. For each, we stochastically modified their corresponding known biologically active small-molecule inhibitors using a linker with a photoactivatable diazirine group. Resulting reaction mixture contained various inhibitor isomers, whose biologic activity was not compromised by the position of the linker attachment as well as those where it was. Afterwards, the reaction mixtures were conjugated to N-(2-hydroxypropyl)methacrylamide copolymers (called iBodies) carrying biotin and a fluorophore, which were used as fully synthetic antibody mimetics. In all cases, stochastic polymer conjugates were able to pull-down the corresponding target protein from cell lysates. For the GCPII conjugate, we then analyzed the pulled-down proteins by mass spectrometry, which clearly identified GCPII as the target protein of the conjugate, even with no prior knowledge needed. For GCPII and FAP, which are cell surface receptors, those same conjugates were able to visualize the proteins on cells both in confocal microscopy and flow cytometry. On a model case of GCPII we decided to identify the position of the “productive” linker attachment so that it would allow rational synthesis for future use. Using HPLC, we separated the reaction mixture into several fractions, each enriched in a specific linker attachment position. As expected, the fractions differed in binding properties compared to the whole mixture. The position of the linker attachment was determined by mass spectrometry for selected active and inactive fractions. The identified linker positions and activity of the compounds corresponded to the known mode of binding of the small molecule into the active site of GCPII. Polymer conjugates prepared from the most favorable fraction had improved binding properties. This “improved” conjugate allowed more efficient isolation and subsequent i
表型筛选的复兴导致生物活性小分子的发现，导致对快速有效地识别其蛋白质靶标的高需求。这通常是通过为目标下拉准备亲和探针来完成的，这需要耗时的结构-活性关系研究，以确定具有必要连接器的亲和锚的连接的适当位置。我们开始完全绕过这一步骤，并准备随机共轭物，这将使快速目标识别不需要事先的结构信息或测试合适的连接物附着位置。我们建议它们也可以用于鉴定表型活性化合物的未知靶点以及已建立的活性化合物的非靶点。为了验证这种新方法，我们选择了几种模型蛋白靶点——在前列腺癌中表达的谷氨酸羧肽酶II (GCPII，也称为前列腺特异性膜抗原，PSMA)、肿瘤基质中的成纤维细胞激活蛋白(FAP)和天门氨酸蛋白酶家族，特别是胃蛋白酶、组织蛋白酶D和HIV蛋白酶。对于每一种，我们随机修改其相应的已知生物活性小分子抑制剂，使用具有光活化重氮嘧啶基团的连接剂。由此产生的反应混合物含有各种抑制剂异构体，其生物活性不受连接物附着位置的影响，也不受连接物附着位置的影响。然后，将反应混合物偶联成携带生物素和荧光团的N-(2-羟丙基)甲基丙烯酰胺共聚物(称为iBodies)，用作完全合成的抗体模拟物。在所有情况下，随机聚合物偶联物都能够从细胞裂解物中拉下相应的靶蛋白。对于GCPII偶联物，我们随后通过质谱分析了拉下蛋白，即使不需要事先了解，也可以清楚地确定GCPII是偶联物的靶蛋白。对于细胞表面受体GCPII和FAP，这些相同的偶联物能够在共聚焦显微镜和流式细胞术中显示细胞上的蛋白质。在GCPII的模型案例中，我们决定确定“生产性”链接附件的位置，以便它可以合理地合成以供将来使用。使用高效液相色谱法，我们将反应混合物分成几个部分，每个部分在特定的连接物附着位置富集。正如预期的那样，与整个混合物相比，这些分数的结合特性不同。对选定的活性和非活性组分，采用质谱法确定连接物附着的位置。所鉴定的化合物的连接位置和活性与已知的小分子结合到GCPII活性位点的模式相对应。由最优馏分制备的聚合物偶联物具有较好的结合性能。这种“改进的”共轭物允许更有效的分离和随后的质谱鉴定GCPII。这些结果表明，生物活性小分子的随机修饰结合iBodies的亲和效应，使我们避免了冗长的结构-活性关系研究，并且可以使用单个偶联物识别蛋白质靶点并将其可视化。该技术可能有助于快速、简便地鉴定具有表型活性的小分子蛋白靶点。本研究由第5号基金资助。GA16-02938S来自捷克共和国资助机构，InterBioMed项目LO 1302来自捷克共和国教育部，查尔斯大学项目GA UK No. 1510-243-250045。引文格式:Kristyna Blazkova, peter Simon, Tomas Knedlik, Petra Dvořakova, Anna Březinova, Libor Kostka, Vladimir Subr, Jan Konvalinka, Pavel Sacha。从具有生物活性的小分子到已鉴定的蛋白质靶标:一种结合随机光电化和合成抗体模拟物的新方法[摘要]。第四届CRI-CIMT-EATI-AACR国际癌症免疫治疗会议:将科学转化为生存;2018年9月30日至10月3日;纽约，纽约。费城(PA): AACR;癌症免疫学杂志2019;7(2增刊):摘要nr B147。
{"title":"Abstract B147: From bioactive small molecule to an identified protein target: A new method combining stochastic photomodification with a synthetic antibody mimetic","authors":"K. Blažková, Petr Šimon, Tomáš Knedlík, P. Dvořáková, A. Březinová, L. Kostka, V. Šubr, J. Konvalinka, P. Šácha","doi":"10.1158/2326-6074.CRICIMTEATIAACR18-B147","DOIUrl":"https://doi.org/10.1158/2326-6074.CRICIMTEATIAACR18-B147","url":null,"abstract":"The renaissance of phenotypic screening leads to the discovery of biologically active small molecules resulting in a high need to quickly and efficiently identify their protein target. This is most commonly done by a preparation of an affinity probe for the target pull-down, which requires time-consuming structure-activity relationship studies to identify proper position for an attachment of an affinity anchor with necessary linker. We set out to circumvent this step altogether and prepare stochastic conjugates that would enable fast target identification without prior structural information or testing of suitable linker attachment position. We suggest that they could be used also for the identification of unknown targets of phenotypically active compounds as well as off-targets of established active compounds. To test the new approach, we selected several model protein targets – glutamate carboxypeptidase II (GCPII, also known as prostate specific membrane antigen, PSMA) expressed in prostate cancer, fibroblast activation protein (FAP) in tumor stroma, and aspartic protease family, specifically pepsin, cathepsin D and HIV protease. For each, we stochastically modified their corresponding known biologically active small-molecule inhibitors using a linker with a photoactivatable diazirine group. Resulting reaction mixture contained various inhibitor isomers, whose biologic activity was not compromised by the position of the linker attachment as well as those where it was. Afterwards, the reaction mixtures were conjugated to N-(2-hydroxypropyl)methacrylamide copolymers (called iBodies) carrying biotin and a fluorophore, which were used as fully synthetic antibody mimetics. In all cases, stochastic polymer conjugates were able to pull-down the corresponding target protein from cell lysates. For the GCPII conjugate, we then analyzed the pulled-down proteins by mass spectrometry, which clearly identified GCPII as the target protein of the conjugate, even with no prior knowledge needed. For GCPII and FAP, which are cell surface receptors, those same conjugates were able to visualize the proteins on cells both in confocal microscopy and flow cytometry. On a model case of GCPII we decided to identify the position of the “productive” linker attachment so that it would allow rational synthesis for future use. Using HPLC, we separated the reaction mixture into several fractions, each enriched in a specific linker attachment position. As expected, the fractions differed in binding properties compared to the whole mixture. The position of the linker attachment was determined by mass spectrometry for selected active and inactive fractions. The identified linker positions and activity of the compounds corresponded to the known mode of binding of the small molecule into the active site of GCPII. Polymer conjugates prepared from the most favorable fraction had improved binding properties. This “improved” conjugate allowed more efficient isolation and subsequent i","PeriodicalId":120683,"journal":{"name":"Other Topics","volume":"39 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2019-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"132458390","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 : 2019-02-01DOI: 10.1158/2326-6074.CRICIMTEATIAACR18-B179
K. Pestal, G. Barton
When normal, healthy cells die by apoptosis, they must be cleared to ensure tissue homeostasis. Failure to clear apoptotic cells (ACs) efficiently enables the release of inflammatory stimuli, such as nucleic acids, leading to tissue injury and, ultimately, autoimmunity. Clearance of ACs is largely performed by tissue-resident macrophages. Despite expressing many innate immune receptors, these cells do not respond to the potentially stimulatory ligands within ACs. Recently, the Barton Lab has identified several populations of AC-engulfing macrophages with many common features. These features include the expression of AC-recognition receptors and reduced TLR responsiveness to nucleic acids. The transcription factors KLF2 and KLF4 appear to contribute to this expression program both in vitro and in vivo in distinct AC-clearing macrophage populations. Macrophages that are removed from their environments lose this program and the program can be adopted by transferring TLR responsive macrophages into environments where endogenous silent residents are maintained. This coordinated silent immune program allows tissue-resident macrophages to maintain homeostasis while efficiently clearing ACs. Citation Format: Kathleen Pestal, Gregory M. Barton. The regulation of silent apoptotic cell-clearance in tissue-resident macrophages [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr B179.
当正常、健康的细胞因凋亡而死亡时，它们必须被清除以确保组织的稳态。如果不能有效清除凋亡细胞(ACs)，就会释放炎症刺激，如核酸，导致组织损伤，最终导致自身免疫。ACs的清除主要是由组织内巨噬细胞完成的。尽管表达了许多先天免疫受体，但这些细胞对ACs内潜在的刺激配体没有反应。最近，Barton实验室已经确定了几种具有许多共同特征的ac吞噬巨噬细胞。这些特征包括交流识别受体的表达和TLR对核酸的反应性降低。转录因子KLF2和KLF4似乎在体外和体内不同的ac清除巨噬细胞群体中都参与了这种表达程序。离开环境的巨噬细胞失去了这一程序，可以通过将TLR反应性巨噬细胞转移到维持内源性沉默居民的环境中来采用这一程序。这种协调的沉默免疫程序允许组织内巨噬细胞在有效清除ACs的同时保持体内平衡。引用格式:Kathleen Pestal, Gregory M. Barton。组织内巨噬细胞沉默凋亡细胞清除的调控[摘要]。第四届CRI-CIMT-EATI-AACR国际癌症免疫治疗会议:将科学转化为生存;2018年9月30日至10月3日;纽约，纽约。费城(PA): AACR;癌症免疫学杂志，2019;7(2增刊):摘要nr B179。
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Pub Date : 2019-02-01DOI: 10.1158/2326-6074.CRICIMTEATIAACR18-B144
B. Azhken
In this case, the process of development of the Barrett9s esophagus and the development of adenocarcinoma as a complication of this disease with involvement of lymphoid tissue with the formation of follicles of different types was studied. Barrett9s esophagus (BE) is a precursor of adenocarcinoma of the esophagus, a disease with a growing burden in the Western world. The incidence of BE increased dramatically during the late 20th century and increased morbidity of this disease. Prevalence is between 0.5 and 2.0%. There are risk factors for BE, including obesity and tobacco smoking, but gastroesophageal reflux disease (GERD) is the strongest risk factor. Adenocarcinoma of the esophagus (ACE) is the most common form of esophageal cancer. However, the data from United Kingdom and the Netherlands show that the incidence of BE increased even after monitoring the increase in endoscopy indices. These estimates indicate an increase in the incidence of BE in approximately 65% between 1997 and 2002 and 159% between 1993 and 2005. It is disturbing that the greatest proportionate increase in the diagnosis of BE was in people younger than 60 years, which is consistent with other works from Europe. In analyzing the literature relating to the intrinsic glands of the esophagus and Barrett esophagus, concerned the esophagus9s own glands and the presence of a large number of endocrine cells in them. N.I. Kolycheva, R.R. Bektayeva studied at ultramicroscopic level. When studying the work on the lymphoid formations of the esophagus, we can cite, concerning the lymphoid component of the esophagus. At present, the source of lymphocyte infiltration into its proper mucous plate, types of lymphoid formations, has not been studied. To study the lymphoid noncapsular formations of the mucous membrane of Barrett9s esophagus, biopsy specimens of Barrett9s esophagus were taken. Early only the epithelium of the esophagus was studied, but there was no evidence of a propria of the mucous membrane and lymphoid tissue. Citation Format: Bakhtiyar Talgatuly Azhken. Morphologic consequences for noncapsular lymphoid tissue in the case of malignancy of GERD (Barrett9s esophagus) [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr B144.
{"title":"Abstract B144: Morphologic consequences for noncapsular lymphoid tissue in the case of malignancy of GERD (Barrett's esophagus)","authors":"B. Azhken","doi":"10.1158/2326-6074.CRICIMTEATIAACR18-B144","DOIUrl":"https://doi.org/10.1158/2326-6074.CRICIMTEATIAACR18-B144","url":null,"abstract":"In this case, the process of development of the Barrett9s esophagus and the development of adenocarcinoma as a complication of this disease with involvement of lymphoid tissue with the formation of follicles of different types was studied. Barrett9s esophagus (BE) is a precursor of adenocarcinoma of the esophagus, a disease with a growing burden in the Western world. The incidence of BE increased dramatically during the late 20th century and increased morbidity of this disease. Prevalence is between 0.5 and 2.0%. There are risk factors for BE, including obesity and tobacco smoking, but gastroesophageal reflux disease (GERD) is the strongest risk factor. Adenocarcinoma of the esophagus (ACE) is the most common form of esophageal cancer. However, the data from United Kingdom and the Netherlands show that the incidence of BE increased even after monitoring the increase in endoscopy indices. These estimates indicate an increase in the incidence of BE in approximately 65% between 1997 and 2002 and 159% between 1993 and 2005. It is disturbing that the greatest proportionate increase in the diagnosis of BE was in people younger than 60 years, which is consistent with other works from Europe. In analyzing the literature relating to the intrinsic glands of the esophagus and Barrett esophagus, concerned the esophagus9s own glands and the presence of a large number of endocrine cells in them. N.I. Kolycheva, R.R. Bektayeva studied at ultramicroscopic level. When studying the work on the lymphoid formations of the esophagus, we can cite, concerning the lymphoid component of the esophagus. At present, the source of lymphocyte infiltration into its proper mucous plate, types of lymphoid formations, has not been studied. To study the lymphoid noncapsular formations of the mucous membrane of Barrett9s esophagus, biopsy specimens of Barrett9s esophagus were taken. Early only the epithelium of the esophagus was studied, but there was no evidence of a propria of the mucous membrane and lymphoid tissue. Citation Format: Bakhtiyar Talgatuly Azhken. Morphologic consequences for noncapsular lymphoid tissue in the case of malignancy of GERD (Barrett9s esophagus) [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr B144.","PeriodicalId":120683,"journal":{"name":"Other Topics","volume":"18 12","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2019-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"121003413","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 : 2019-02-01DOI: 10.1158/2326-6074.CRICIMTEATIAACR18-B164
Shi-Wei Hunag, J. Shieh, Sin-Ting Wang, D. Cho, S. Chiu
Immunogenic cell death (ICD) characterized as cells undergo with ICD could elicit a specific immune response to against a subsequent challenge with the same type living cells. Imiquimod (IMQ), a synthetic Toll-like receptor 7 ligand, is a imidazoquinoline family member and contains both antitumor and antiviral activity. IMQ is currently clinical used in topical, noninvasive treatment for various skin malignances. IMQ exerts its anti-tumoral activity through stimulating TLR7/8 in dendritic cells and activate cell-mediated immune responses, and directly induces tumor cell death. Recently, we had demonstrated IMQ directly induced autophagic cell death and apoptosis to against tumor cells that independent of TLR7/8. Thus, IMQ may induce tumor undergo ICD to trigger specific antitumor immune response. We found that IMQ induced expression of ICD-associated features in various cancer cell lines. We demonstrated that vaccination of IMQ-induced ICD cell lysate provided significant antitumor immunity with increased CD4 and CD8 T lymphocytes proliferation, tumor-specific cytotoxic lysis and infiltration of CD4, CD8, CD11c and IL-17A-producing cells in tumor lesion. Pharmacologic inhibition and genetic manipulation of ICD-associated features repressed IMQ-treated cell lysate-mediated specific anti-tumor immunity. We also found that IMQ-induced ICD-associated features is mediated by ROS production and ER stress through PERK/eIF2α pathway. In addition, inhibition of glycolysis significantly enhanced IMQ-induced ICD cell lysate-mediated antitumor immunity and ICD-associated features. Taken together, these results provided solid evidence to support that IMQ is an authentic ICD inducer and independent of TLR7/8 expression, and targeting aerobic glycolysis could be a desirable strategy to enhance IMQ-induced ICD and antitumor immunity. Citation Format: Shi-Wei Hunag, Jeng-Jer Shieh, Sin-Ting Wang, Der-Yang Cho, Shao-Chih Chiu. Inhibiting glycolysis enhances imiquimod-induced immunogenic cell death and the antitumor immune response [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr B164.
{"title":"Abstract B164: Inhibiting glycolysis enhances imiquimod-induced immunogenic cell death and the antitumor immune response","authors":"Shi-Wei Hunag, J. Shieh, Sin-Ting Wang, D. Cho, S. Chiu","doi":"10.1158/2326-6074.CRICIMTEATIAACR18-B164","DOIUrl":"https://doi.org/10.1158/2326-6074.CRICIMTEATIAACR18-B164","url":null,"abstract":"Immunogenic cell death (ICD) characterized as cells undergo with ICD could elicit a specific immune response to against a subsequent challenge with the same type living cells. Imiquimod (IMQ), a synthetic Toll-like receptor 7 ligand, is a imidazoquinoline family member and contains both antitumor and antiviral activity. IMQ is currently clinical used in topical, noninvasive treatment for various skin malignances. IMQ exerts its anti-tumoral activity through stimulating TLR7/8 in dendritic cells and activate cell-mediated immune responses, and directly induces tumor cell death. Recently, we had demonstrated IMQ directly induced autophagic cell death and apoptosis to against tumor cells that independent of TLR7/8. Thus, IMQ may induce tumor undergo ICD to trigger specific antitumor immune response. We found that IMQ induced expression of ICD-associated features in various cancer cell lines. We demonstrated that vaccination of IMQ-induced ICD cell lysate provided significant antitumor immunity with increased CD4 and CD8 T lymphocytes proliferation, tumor-specific cytotoxic lysis and infiltration of CD4, CD8, CD11c and IL-17A-producing cells in tumor lesion. Pharmacologic inhibition and genetic manipulation of ICD-associated features repressed IMQ-treated cell lysate-mediated specific anti-tumor immunity. We also found that IMQ-induced ICD-associated features is mediated by ROS production and ER stress through PERK/eIF2α pathway. In addition, inhibition of glycolysis significantly enhanced IMQ-induced ICD cell lysate-mediated antitumor immunity and ICD-associated features. Taken together, these results provided solid evidence to support that IMQ is an authentic ICD inducer and independent of TLR7/8 expression, and targeting aerobic glycolysis could be a desirable strategy to enhance IMQ-induced ICD and antitumor immunity. Citation Format: Shi-Wei Hunag, Jeng-Jer Shieh, Sin-Ting Wang, Der-Yang Cho, Shao-Chih Chiu. Inhibiting glycolysis enhances imiquimod-induced immunogenic cell death and the antitumor immune response [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr B164.","PeriodicalId":120683,"journal":{"name":"Other Topics","volume":"98 4 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2019-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"116403357","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 : 2019-02-01DOI: 10.1158/2326-6074.CRICIMTEATIAACR18-B198
Hao Xu, D. Littman, W. Huang
RORγt plays vital roles in development and differentiation of multiple immune cell types with pleiotropic functions, including thymocytes, LTi cells, ILC3, γδ T-cells and Th17 cells. Moreover, RORγ, the isoform of RORγt, was reported to promote cancer cell survival in castration-resistant prostate cancer. RORγ is expressed in tissues like muscle, liver and brain, but its function remains unclear. To address the mechanisms by which RORγt/RORγ contributes to diverse cell fates and differentiation programs, we are exploring RORγt post-translational modifications, RORγt/RORγ-associated protein complexes and downstream signals. To identify RORγt/RORγ-associated protein complexes, we generated knock-in mice expressing Strep-tagged RORγt/RORγ, which had IL-17A inducing activity similar to that of non-tagged RORγt in Th17 cells polarized in vitro and allowed for highly specific isolation of protein complexes. Strep-tagged RORγt proteins will be precipitated from thymocytes and polarized pathogenic or nonpathogenic Th17 cells, followed by mass-spectrometry to identify RORγt-associated protein partners and analysis of their functions in RORγt expressing cells. Moreover, due to its nonoverlapping expression patterns with RORγt and obscure functions, RORγ is very interesting to investigate in terms of associated protein complexes and roles in other tissues. RORγt mutations were employed to dissect its roles in cell fate determination and differentiation. We identified a RORγt mutant by random mutagenesis, designated as MUT, which dramatically lost transcriptional activity when expressed in 293T-cells and in vitro polarized Rorc-KO Th17 cells. Mice harboring MUT mutation had normal thymocyte differentiation and development of secondary lymphoid tissues. No lymphomas were detected in MUT mice, unlike in C57BL/6 RORγt-KO mice that develop T-cell lymphomas. Strikingly, T-cells and ILCs differed in proportions and numbers according to tissues. In secondary lymphoid organs, including draining lymph nodes and spleen, no differences were detected in proportions or numbers of CD4 or CD8 T-cells. In large intestine, however, the proportions and numbers of γδ T-cells and Th17 cells were significantly reduced in MUT mice, while those of Th2 cells were markedly increased. Meanwhile, hyper-proliferation of RORγt-negative T helper cells and Treg cells was detected in large intestine of MUT mice. ILCs from large intestine, including ILC3 with high expression of RORγt, were similar to wild type mice in both proportions and numbers. A different phenotype was detected in small intestine, where T-cells and ILCs were markedly reduced. Given the different requirement or expression stage of RORγt for development of T-cells and ILCs, the results probably reflect the effects of RORγt-associated functions on microenvironments. Analyses of immune cells and microbiome at different ages may help to address the mechanism. The functions of MUT RORγt-expressing immune cells will be explore
{"title":"Abstract B198: Dissection of RORγt functions in lymphoid cell fate determination and differentiation","authors":"Hao Xu, D. Littman, W. Huang","doi":"10.1158/2326-6074.CRICIMTEATIAACR18-B198","DOIUrl":"https://doi.org/10.1158/2326-6074.CRICIMTEATIAACR18-B198","url":null,"abstract":"RORγt plays vital roles in development and differentiation of multiple immune cell types with pleiotropic functions, including thymocytes, LTi cells, ILC3, γδ T-cells and Th17 cells. Moreover, RORγ, the isoform of RORγt, was reported to promote cancer cell survival in castration-resistant prostate cancer. RORγ is expressed in tissues like muscle, liver and brain, but its function remains unclear. To address the mechanisms by which RORγt/RORγ contributes to diverse cell fates and differentiation programs, we are exploring RORγt post-translational modifications, RORγt/RORγ-associated protein complexes and downstream signals. To identify RORγt/RORγ-associated protein complexes, we generated knock-in mice expressing Strep-tagged RORγt/RORγ, which had IL-17A inducing activity similar to that of non-tagged RORγt in Th17 cells polarized in vitro and allowed for highly specific isolation of protein complexes. Strep-tagged RORγt proteins will be precipitated from thymocytes and polarized pathogenic or nonpathogenic Th17 cells, followed by mass-spectrometry to identify RORγt-associated protein partners and analysis of their functions in RORγt expressing cells. Moreover, due to its nonoverlapping expression patterns with RORγt and obscure functions, RORγ is very interesting to investigate in terms of associated protein complexes and roles in other tissues. RORγt mutations were employed to dissect its roles in cell fate determination and differentiation. We identified a RORγt mutant by random mutagenesis, designated as MUT, which dramatically lost transcriptional activity when expressed in 293T-cells and in vitro polarized Rorc-KO Th17 cells. Mice harboring MUT mutation had normal thymocyte differentiation and development of secondary lymphoid tissues. No lymphomas were detected in MUT mice, unlike in C57BL/6 RORγt-KO mice that develop T-cell lymphomas. Strikingly, T-cells and ILCs differed in proportions and numbers according to tissues. In secondary lymphoid organs, including draining lymph nodes and spleen, no differences were detected in proportions or numbers of CD4 or CD8 T-cells. In large intestine, however, the proportions and numbers of γδ T-cells and Th17 cells were significantly reduced in MUT mice, while those of Th2 cells were markedly increased. Meanwhile, hyper-proliferation of RORγt-negative T helper cells and Treg cells was detected in large intestine of MUT mice. ILCs from large intestine, including ILC3 with high expression of RORγt, were similar to wild type mice in both proportions and numbers. A different phenotype was detected in small intestine, where T-cells and ILCs were markedly reduced. Given the different requirement or expression stage of RORγt for development of T-cells and ILCs, the results probably reflect the effects of RORγt-associated functions on microenvironments. Analyses of immune cells and microbiome at different ages may help to address the mechanism. The functions of MUT RORγt-expressing immune cells will be explore","PeriodicalId":120683,"journal":{"name":"Other Topics","volume":"44-46 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2019-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123640849","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 : 2019-02-01DOI: 10.1158/2326-6074.CRICIMTEATIAACR18-B201
Ryan Zander, David M. Schauder, G. Xin, C. Nguyen, Xiaopeng Wu, W. Cui
Macrophages can impact the development of tumors and their responses to therapy. However, we lack a full understanding of how macrophage activation and survival are regulated and might be therapeutically modulated. To address these questions, we turned to a host-pathogen interaction that is likely to have impacted the evolution of these processes: infection by Mycobacterium tuberculosis (Mtb), the leading cause of death from infection. Once internalized by macrophages, Mtb resists killing by macrophages and replicates within them. Ultimately, Mtb infection results in death of macrophages, allowing dissemination of the pathogen to other cells. To explore how Mtb induces macrophage cell death and how macrophage cell death might impact host defense against Mtb, we performed a genome-wide CRISPR-Cas9 recessive genetic screen in RAW264.7 macrophages. We discovered that the absence of components of the type I interferon signaling pathway, including the IFN-α/β receptor (IFNAR), delays Mtb-induced cell death. It is known that Mtb infection induces macrophages to produce type I interferon. Our finding directly links type I interferon signaling pathway and Mtb-induced macrophage death. We are currently working on the mechanism of the type I IFN-induced death of bacterially-stimulated macrophages. Meanwhile, we are testing blockers of this pathway as host-directed therapy against TB and have seen striking protective effects of anti-IFNAR1 mAb in Mtb-infected mice, whether the mAb is administered before or after infection. Further exploration of the protective effect of anti-IFNAR mAb could point to an antibody-based, host-directed therapy. Acknowledgment: We thank Prof. R. Schreiber, Washington University, for facilitating access to anti-IFNAR mAb. Citation Format: Ryan Zander, David Schauder, Gang Xin, Christine Nguyen, Xiaopeng Wu, Weiguo Cui. Exploration and exploitation of macrophage death pathways: Infection by Mycobacterium tuberculosis as a model [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr B201.
巨噬细胞可以影响肿瘤的发展及其对治疗的反应。然而，我们对巨噬细胞的激活和存活是如何被调节的以及如何被治疗性地调节缺乏充分的了解。为了解决这些问题，我们转向了可能影响这些过程演变的宿主-病原体相互作用:结核分枝杆菌(Mtb)感染，这是感染导致死亡的主要原因。一旦被巨噬细胞内化，结核分枝杆菌抵抗巨噬细胞的杀伤并在巨噬细胞内复制。最终，结核分枝杆菌感染导致巨噬细胞死亡，使病原体传播到其他细胞。为了探索结核分枝杆菌如何诱导巨噬细胞死亡以及巨噬细胞死亡如何影响宿主对结核分枝杆菌的防御，我们在RAW264.7巨噬细胞中进行了全基因组CRISPR-Cas9隐性遗传筛选。我们发现I型干扰素信号通路成分的缺失，包括IFN-α/β受体(IFNAR)，延迟了mmb诱导的细胞死亡。已知结核分枝杆菌感染诱导巨噬细胞产生I型干扰素。我们的发现将I型干扰素信号通路与mmb诱导的巨噬细胞死亡直接联系起来。我们目前正在研究I型ifn诱导细菌刺激的巨噬细胞死亡的机制。与此同时，我们正在测试这一途径的阻断剂作为针对结核病的宿主定向治疗，并在mtb感染的小鼠中发现了抗ifnar1单抗的显著保护作用，无论该单抗是在感染之前还是之后给药。进一步探索抗ifnar单抗的保护作用可能会指向一种基于抗体的宿主定向治疗。感谢:我们感谢华盛顿大学R. Schreiber教授为获得抗ifnar单抗提供了便利。引文格式:Ryan Zander, David Schauder，辛刚，Christine Nguyen，吴晓鹏，崔卫国。巨噬细胞死亡途径的探索与开发:以结核分枝杆菌感染为模型[摘要]。第四届CRI-CIMT-EATI-AACR国际癌症免疫治疗会议:将科学转化为生存;2018年9月30日至10月3日;纽约，纽约。费城(PA): AACR;癌症免疫学杂志，2019;7(2增刊):摘要nr B201。
{"title":"Abstract B201: Exploration and exploitation of macrophage death pathways: Infection by Mycobacterium tuberculosis as a model","authors":"Ryan Zander, David M. Schauder, G. Xin, C. Nguyen, Xiaopeng Wu, W. Cui","doi":"10.1158/2326-6074.CRICIMTEATIAACR18-B201","DOIUrl":"https://doi.org/10.1158/2326-6074.CRICIMTEATIAACR18-B201","url":null,"abstract":"Macrophages can impact the development of tumors and their responses to therapy. However, we lack a full understanding of how macrophage activation and survival are regulated and might be therapeutically modulated. To address these questions, we turned to a host-pathogen interaction that is likely to have impacted the evolution of these processes: infection by Mycobacterium tuberculosis (Mtb), the leading cause of death from infection. Once internalized by macrophages, Mtb resists killing by macrophages and replicates within them. Ultimately, Mtb infection results in death of macrophages, allowing dissemination of the pathogen to other cells. To explore how Mtb induces macrophage cell death and how macrophage cell death might impact host defense against Mtb, we performed a genome-wide CRISPR-Cas9 recessive genetic screen in RAW264.7 macrophages. We discovered that the absence of components of the type I interferon signaling pathway, including the IFN-α/β receptor (IFNAR), delays Mtb-induced cell death. It is known that Mtb infection induces macrophages to produce type I interferon. Our finding directly links type I interferon signaling pathway and Mtb-induced macrophage death. We are currently working on the mechanism of the type I IFN-induced death of bacterially-stimulated macrophages. Meanwhile, we are testing blockers of this pathway as host-directed therapy against TB and have seen striking protective effects of anti-IFNAR1 mAb in Mtb-infected mice, whether the mAb is administered before or after infection. Further exploration of the protective effect of anti-IFNAR mAb could point to an antibody-based, host-directed therapy. Acknowledgment: We thank Prof. R. Schreiber, Washington University, for facilitating access to anti-IFNAR mAb. Citation Format: Ryan Zander, David Schauder, Gang Xin, Christine Nguyen, Xiaopeng Wu, Weiguo Cui. Exploration and exploitation of macrophage death pathways: Infection by Mycobacterium tuberculosis as a model [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr B201.","PeriodicalId":120683,"journal":{"name":"Other Topics","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2019-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"128863777","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 : 2018-09-01DOI: 10.1158/2326-6074.TUMIMM17-A75
B. Jang, I. Kim
Background: Identifying predictive factors for the clinical outcome of patients with low grade gliomas following radiotherapy could help optimize patient treatments. Here, we investigate the predictive efficacy of both a previously identified “31 gene signature” and programmed death ligand-1 (PD-L1) expression. Methods: We identified 516 patients with low grade glioma in The Cancer Genome Atlas dataset and divided them into two clusters: radiosensitive (RS) and radioresistant (RR). Patients were also classified as PD-L1-high or PD-L1-low based on CD274 mRNA expression. Five-year survival rates were compared across patient groups, and differentially expressed genes were identified via a gene enrichment analysis. Results: Patients in the RS group had a higher survival rate than in the RR group, but only when treated with radiotherapy (63% vs. 52%, p = 0.019; univariate analysis). Patients in the RR group who were also PD-L1-high had a lower survival rate than other patients (50% vs. 60%, p = 0.010). Furthermore, a Cox multivariate analysis demonstrated that both belonging to this patient population and expressing wild-type isocitrate dehydrogenase were predictive of lower survival rates (p = 0.048). Differentially expressed genes associated with this group were found to play a role in the immune response, including the T-cell receptor signaling pathway. Conclusions: We validated the predictive value of a 31 gene signature and PD-L1 expression in a dataset of patients with low-grade glioma. Our results suggest that the patient population classified as both RR and PD-L1 high may benefit most from radiotherapy combined with anti-PD1/PD-L1 treatment. This work was supported by a grant from the Ministry of Science, ICT & Future Planning (NRF#2017R1A2B4002710 & & 2017M2A2A7A01018438) to In Ah Kim. Citation Format: Bum Sup Jang, In Ah Kim. A radiosensitivity gene signature and PD-L1 expression are predictive of the clinical outcomes of patients with lower-grade glioma in The Cancer Genome Atlas dataset [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2017 Oct 1-4; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2018;6(9 Suppl):Abstract nr A75.
{"title":"Abstract A75: A radiosensitivity gene signature and PD-L1 expression are predictive of the clinical outcomes of patients with lower-grade glioma in The Cancer Genome Atlas dataset","authors":"B. Jang, I. Kim","doi":"10.1158/2326-6074.TUMIMM17-A75","DOIUrl":"https://doi.org/10.1158/2326-6074.TUMIMM17-A75","url":null,"abstract":"Background: Identifying predictive factors for the clinical outcome of patients with low grade gliomas following radiotherapy could help optimize patient treatments. Here, we investigate the predictive efficacy of both a previously identified “31 gene signature” and programmed death ligand-1 (PD-L1) expression. Methods: We identified 516 patients with low grade glioma in The Cancer Genome Atlas dataset and divided them into two clusters: radiosensitive (RS) and radioresistant (RR). Patients were also classified as PD-L1-high or PD-L1-low based on CD274 mRNA expression. Five-year survival rates were compared across patient groups, and differentially expressed genes were identified via a gene enrichment analysis. Results: Patients in the RS group had a higher survival rate than in the RR group, but only when treated with radiotherapy (63% vs. 52%, p = 0.019; univariate analysis). Patients in the RR group who were also PD-L1-high had a lower survival rate than other patients (50% vs. 60%, p = 0.010). Furthermore, a Cox multivariate analysis demonstrated that both belonging to this patient population and expressing wild-type isocitrate dehydrogenase were predictive of lower survival rates (p = 0.048). Differentially expressed genes associated with this group were found to play a role in the immune response, including the T-cell receptor signaling pathway. Conclusions: We validated the predictive value of a 31 gene signature and PD-L1 expression in a dataset of patients with low-grade glioma. Our results suggest that the patient population classified as both RR and PD-L1 high may benefit most from radiotherapy combined with anti-PD1/PD-L1 treatment. This work was supported by a grant from the Ministry of Science, ICT & Future Planning (NRF#2017R1A2B4002710 & & 2017M2A2A7A01018438) to In Ah Kim. Citation Format: Bum Sup Jang, In Ah Kim. A radiosensitivity gene signature and PD-L1 expression are predictive of the clinical outcomes of patients with lower-grade glioma in The Cancer Genome Atlas dataset [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2017 Oct 1-4; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2018;6(9 Suppl):Abstract nr A75.","PeriodicalId":120683,"journal":{"name":"Other Topics","volume":"29 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2018-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"121973964","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 : 2018-09-01DOI: 10.1158/1557-3265.AACRIASLC18-A10
Miloud Taki Eddine Aichour, Z. Zaidi
Background: Lung cancer is the leading cause of cancer death among men and the second leading cause of cancer death among women worldwide. Rates are highest in countries where smoking uptake began earliest, such as those in North America and Europe. Although rates are now decreasing in most of these countries, especially in men, they are increasing in countries where smoking uptake occurred later. Variation between countries may reflect different prevalence of risk factors. Lung cancer is the leading cause of cancer death among both men and women in the United States. In the United States, smoking rates among women peaked after those among men. The main objective is to estimate age-standardized mortality rates by U.S. county from tracheal, bronchus, and lung cancer. Methods: The finding of the Global Burden of Disease (GBD) 2015 methodology with death records from the National Center for Health Statistics (NCHS) and population counts from the Census Bureau, the NCHS, and the Human Mortality Database from 1980 to 2014 were used. Results: A total of 5,656,423 deaths from tracheal, bronchus, and lung (TBL) cancer were recorded. There were large differences in the mortality rate among counties throughout this period. TBL cancer mortality declined by 21.0% (95% UI, 17.9%-24.0%) between 1980 and 2014, from 68.6 (95% UI, 66.8-70.3) deaths per 100,000 population to 54.2 (95% UI, 52.7-55.6). The West and Northeast experienced declines in the mortality rate, as did Florida, while increases were observed in the South, Appalachian region. The largest increase from 1980 to 2014 was observed in Owsley County, Kentucky (99.7%; 95% UI, 73.7%-130.8%), while the greatest decline was observed in Aleutians East Borough and Aleutians West Census Area, Alaska (63.6%; 95% UI, 50.3%-73.5%). High mortality rates in 2014 were clustered in West Virginia. Because national rates peaked in 1988, women in 2,215 counties experienced a statistically significant increase in the mortality rate, while this was true for men in only 11 counties. The highest national mortality rate for men was present in 1980, while the peak in mortality rate for women was in 2001. The largest percentage increase (168.3%; 95% UI, 136.4%207.8) from 1980 to the peak in 2001 for women was observed in Marlboro County, South Carolina (mortality rate of 67.1 deaths per 100 000). Mortality rates varied from 10.6 (95% UI, 8.6-12.8) in Summit County, Colorado, to 334.9 (95% UI, 300.5-375.2) in Union County, Florida, for males and 10.9 (95% UI, 8.3-13.8) in Summit County, Colorado, to 121 (95% UI, 101.6-142.0) in Owsley County, Kentucky, for females. Low rates were observed along the US border with Mexico and in Utah, Colorado, and parts of Arizona, New Mexico, and Idaho. Conclusion: From 1980 to 2014 there has been a steady decline in the cancer death rate as a result of fewer Americans smoking and advances in cancer prevention, early detection, and treatment. Local efforts to reduce smoking in poor and rural
{"title":"Abstract A10: Trends and patterns of disparities in tracheal, bronchus, and lung cancer mortality among U.S. counties, 1980-2014","authors":"Miloud Taki Eddine Aichour, Z. Zaidi","doi":"10.1158/1557-3265.AACRIASLC18-A10","DOIUrl":"https://doi.org/10.1158/1557-3265.AACRIASLC18-A10","url":null,"abstract":"Background: Lung cancer is the leading cause of cancer death among men and the second leading cause of cancer death among women worldwide. Rates are highest in countries where smoking uptake began earliest, such as those in North America and Europe. Although rates are now decreasing in most of these countries, especially in men, they are increasing in countries where smoking uptake occurred later. Variation between countries may reflect different prevalence of risk factors. Lung cancer is the leading cause of cancer death among both men and women in the United States. In the United States, smoking rates among women peaked after those among men. The main objective is to estimate age-standardized mortality rates by U.S. county from tracheal, bronchus, and lung cancer. Methods: The finding of the Global Burden of Disease (GBD) 2015 methodology with death records from the National Center for Health Statistics (NCHS) and population counts from the Census Bureau, the NCHS, and the Human Mortality Database from 1980 to 2014 were used. Results: A total of 5,656,423 deaths from tracheal, bronchus, and lung (TBL) cancer were recorded. There were large differences in the mortality rate among counties throughout this period. TBL cancer mortality declined by 21.0% (95% UI, 17.9%-24.0%) between 1980 and 2014, from 68.6 (95% UI, 66.8-70.3) deaths per 100,000 population to 54.2 (95% UI, 52.7-55.6). The West and Northeast experienced declines in the mortality rate, as did Florida, while increases were observed in the South, Appalachian region. The largest increase from 1980 to 2014 was observed in Owsley County, Kentucky (99.7%; 95% UI, 73.7%-130.8%), while the greatest decline was observed in Aleutians East Borough and Aleutians West Census Area, Alaska (63.6%; 95% UI, 50.3%-73.5%). High mortality rates in 2014 were clustered in West Virginia. Because national rates peaked in 1988, women in 2,215 counties experienced a statistically significant increase in the mortality rate, while this was true for men in only 11 counties. The highest national mortality rate for men was present in 1980, while the peak in mortality rate for women was in 2001. The largest percentage increase (168.3%; 95% UI, 136.4%207.8) from 1980 to the peak in 2001 for women was observed in Marlboro County, South Carolina (mortality rate of 67.1 deaths per 100 000). Mortality rates varied from 10.6 (95% UI, 8.6-12.8) in Summit County, Colorado, to 334.9 (95% UI, 300.5-375.2) in Union County, Florida, for males and 10.9 (95% UI, 8.3-13.8) in Summit County, Colorado, to 121 (95% UI, 101.6-142.0) in Owsley County, Kentucky, for females. Low rates were observed along the US border with Mexico and in Utah, Colorado, and parts of Arizona, New Mexico, and Idaho. Conclusion: From 1980 to 2014 there has been a steady decline in the cancer death rate as a result of fewer Americans smoking and advances in cancer prevention, early detection, and treatment. Local efforts to reduce smoking in poor and rural ","PeriodicalId":120683,"journal":{"name":"Other Topics","volume":"37 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2018-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"133440785","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}
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