Pilot Project 2

试点项目2

基本信息

批准号：
9762871
负责人：
Gayathri Devi
金额：
$ 2.38万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至 2021-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9762871
关键词：
Adult African American Age Apoptosis Archives Automobile Driving Biological Biological Assay Breast Cancer Cell Breast Cancer Model Breast Cancer Patient Breast Cancer cell line CRISPR/Cas technology Cancer Biology Cancer Control Cancer Patient Cancer cell line Cell Line Cell Proliferation Cell model Cells Cellular biology Cessation of life Clinic Collaborations Data Development Disadvantaged Disease Down-Regulation Drug resistance Embryonic Development Erinaceidae Exhibits FRAP1 gene GLI gene Gene Amplification Gene Expression Gene Expression Profile Gene Expression Profiling Genes Genetic Genetic Transcription Goals Growth Human Immunohistochemistry Incidence Income Joints Knock-out Lead Link Malignant Neoplasms Mediating Minority Mission Modeling Molecular Outcome Pathway interactions Patients Pharmacology Phenotype Pilot Projects Population-Based Registry Postdoctoral Fellow Prevalence Public Health Quantitative Reverse Transcriptase PCR Race Relapse Reporting Research Reverse Transcriptase Polymerase Chain Reaction Role Sampling Signal Transduction Testing Therapeutic Time Tissue Sample Translating Translational Research Underrepresented Minority United States National Institutes of Health Validation Woman Work Xenograft procedure advanced breast cancer anticancer research cancer health disparity cancer therapy cell growth cell motility clinical practice differential expression genetic signature genetically modified cells health care availability inflammatory breast cancer inhibitor/antagonist innovation insight knock-down malignant breast neoplasm migration mortality novel novel strategies novel therapeutic intervention novel therapeutics programs repository response screening small hairpin RNA therapeutic target transcription factor transcriptome sequencing tumor tumorigenesis

Adult African American Age Apoptosis Archives Automobile Driving Biological Biological Assay Breast Cancer Cell Breast Cancer Model Breast Cancer Patient Breast Cancer cell line CRISPR/Cas technology Cancer Biology Cancer Control Cancer Patient Cancer cell line Cell Line Cell Proliferation Cell model Cells Cellular biology Cessation of life Clinic Collaborations Data Development Disadvantaged Disease Down-Regulation Drug resistance Embryonic Development Erinaceidae Exhibits FRAP1 gene GLI gene Gene Amplification Gene Expression Gene Expression Profile Gene Expression Profiling Genes Genetic Genetic Transcription Goals Growth Human Immunohistochemistry Incidence Income Joints Knock-out Lead Link Malignant Neoplasms Mediating Minority Mission Modeling Molecular Outcome Pathway interactions Patients Pharmacology Phenotype Pilot Projects Population-Based Registry Postdoctoral Fellow Prevalence Public Health Quantitative Reverse Transcriptase PCR Race Relapse Reporting Research Reverse Transcriptase Polymerase Chain Reaction Role Sampling Signal Transduction Testing Therapeutic Time Tissue Sample Translating Translational Research Underrepresented Minority United States National Institutes of Health Validation Woman Work Xenograft procedure advanced breast cancer anticancer research cancer health disparity cancer therapy cell growth cell motility clinical practice differential expression genetic signature genetically modified cells health care availability inflammatory breast cancer inhibitor/antagonist innovation insight knock-down malignant breast neoplasm migration mortality novel novel strategies novel therapeutic intervention novel therapeutics programs repository response screening small hairpin RNA therapeutic target transcription factor transcriptome sequencing tumor tumorigenesis

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项目摘要

PROJECT ABSTRACT Inflammatory breast cancer (IBC), one of the most lethal forms of breast cancer, accounts for ~15% of all breast cancer deaths. IBC is particularly devastating in disadvantaged minority women who have both a higher incidence and poorer survival. Gene expression profiling of IBC tumors has identified the hedgehog(Hh)/GLI1 pathway as higher in patients with aggressive and relapsing IBC. We have previously demonstrated that GLI1 is highly expressed in IBC cells derived from an aggressive primary IBC tumor, and that down-regulation of GLI1 in these cells decreased cell proliferation, increased apoptosis and reduced migration. The objective of the proposed research is to identify those genes that are critical for GLI activation and function, including those that are both up- and downstream in of GLI1, to provide insight into the molecular mechanisms underlying or driving IBC. The hypothesis to be explored is that GLI1 and the genes that it regulates promote the growth and survival of IBC, and that identifying these genes may lead to the elucidation of novel targets. The long-term goal is to inform novel approaches for anti-tumor efficacy in IBC. This objective will be met by applying an innovative approach that includes utilization of high-throughput transcriptome sequencing (RNA-Seq), genetically engineered cell lines and human IBC tissue samples. This will become accomplished by the following Aims: 1) To identify gene expression differences between low and high GLI1 expressing IBC models including drug resistant and GLI1-antagonist treated IBC models. Differences in gene expression patterns will be determined by high-throughput transcriptome sequencing (RNA-Seq). 2) To confirm and validate those gene expression differences by qRT-PCR analysis including a comparison of IBC and non-IBC cell lines. To assess overlap of those differentially expressed genes with African American patient derived non-IBC cell lines. We will assess GLI1 expression and the expression of a subset of differentially expressed target genes identified in Aim 1 in archival IBC patient and control tissue samples accessed from the Duke Biospecimen Repository and Processing Core by immuno-histochemistry, and gene expression analysis. To define functional roles for these target genes, a subset will be knocked down by shRNA or knocked out using CRISPR/Cas9 and effects on IBC growth and survival will be determined. 3) Selective pharmacological inhibitors will be used to assess the role of previously identified pathways in activating GLI1 in IBC and non- IBC. We assess effects on IBC cell biology in a panel of functional and phenotypic assays. This proposed research will further the collaboration between Duke and NCCU on IBC and will support the development of an under-represented minority postdoctoral fellow performing research focused on transcriptional profiling of GLI1 activation pathways in IBC. Completion of these Aims will provide mechanistic insights into the consequences of GLI1 activation and antagonism in IBC, the downstream pathways involved and inform novel approaches for anti-tumor efficacy in IBC with the long term goal to translate new therapies for IBC into the clinic.

项目摘要炎症性乳腺癌（IBC）是最致命的乳腺癌之一，占所有乳腺癌的15％乳腺癌死亡。 IBC在不利的少数妇女中尤其是毁灭性的发病率和较差的生存。 IBC肿瘤的基因表达分析已鉴定出刺猬（HH）/GLI1 攻击性和复发性IBC患者的途径较高。我们以前已经证明了Gli1 在源自侵略性初级IBC肿瘤的IBC细胞中高度表达这些细胞中的GLI1降低了细胞增殖，凋亡增加并减少迁移。目的拟议的研究是识别那些对GLI激活和功能至关重要的基因，包括在Gli1的上游和下游，以洞悉或驾驶IBC。要探讨的假设是GLI1及其调节的基因促进了生长和 IBC的生存以及鉴定这些基因的生存可能导致阐明新靶标。长期目标是为IBC中的抗肿瘤功效提供新颖的方法。通过应用一个创新方法，包括利用高通量转录组测序（RNA-SEQ），基因设计的细胞系和人类IBC组织样品。这将通过以下目的：1）识别表达IBC模型的低GLI1和高GLI1之间的基因表达差异包括耐药和GLI1抗抗药性的IBC模型。基因表达模式的差异将通过高通量转录组测序（RNA-Seq）确定。 2）确认和验证这些通过QRT-PCR分析进行的基因表达差异，包括比较IBC和非IBC细胞系。到评估与非裔美国人患者衍生的非IBC细胞的那些差异表达基因的重叠线。我们将评估GLI1表达和差异表达靶基因的子集的表达在档案IBC患者的AIM 1中鉴定出来，并控制从Duke BioScecimen访问的组织样本通过免疫 - 归化学和基因表达分析，存储库和处理核心。定义这些靶基因的功能作用，子集将被shRNA击倒或使用 CRISPR/CAS9以及对IBC增长和生存的影响。 3）选择性药理抑制剂将用于评估先前鉴定的途径在激活IBC和非 - 非 - IBC。我们评估对功能和表型测定面板中IBC细胞生物学的影响。这提出了研究将进一步进一步在IBC上与NCCU之间的合作，并将支持开发代表性不足的少数族裔博士后同学表演研究重点是GLI1的转录分析 IBC中的激活途径。这些目标的完成将提供有关后果的机械见解 IBC中Gli1激活和拮抗作用，涉及的下游途径，并为新颖的方法提供了新的方法 IBC的抗肿瘤功效具有长期目标，将IBC的新疗法转化为诊所。