Biomarkers Core

生物标志物核心

• 批准号: 9763462
• 负责人: DAVID S LOOSE
• 金额: $ 4.54万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9763462
• 关键词: Aftercare; Antibodies; Atypical hyperplasia; Biological Assay; Biological Markers; Biopsy; Biopsy Specimen; Blood; CAV1 gene; CTNNB1 gene; Carcinoma; Cell Line; Cells; Clinical; Clinical Trials; Combined Modality Therapy; Complex; DNA; Development; Diagnosis; Disease Resistance; Drug Combinations; Endometrial Carcinoma; Enrollment; Epigenetic Process; Estrogens; Exons; Expression Profiling; Formalin; Gene Chips; Gene Expression; Genes; Gland; Gynecologic; Gynecology; Hormonal; Immunosuppressive Agents; Individual; Liposomes; Malignant Neoplasms; Maps; Mesenchymal; Messenger RNA; MicroRNAs; Microarray Analysis; Microdissection; Molecular; Molecular Profiling; Molecular Target; Mutation; Neurons; Nucleic Acids; Obesity; Paraffin Embedding; Pathway interactions; Patients; Pharmaceutical Preparations; Pharmacology; Pharmacotherapy; Phase; Phosphoproteins; Prevention strategy; Progestins; Protein Array; Protein Array Analysis; Proteins; Proteomics; Protocols documentation; Quantitative Reverse Transcriptase PCR; RNA; Research; Research Personnel; Resistance; Resources; Rodent; SDZ RAD; Sampling; Standardization; Stromal Cells; System; Techniques; Testing; Time; Tissue Embedding; Tissues; Transcript; Treatment Efficacy; Uterine Cancer; Uterus; Validation; base; candidate marker; career; combinatorial; design; differential expression; dimer; experimental study; follow-up; genome-wide; imager; in vitro Model; inhibitor/antagonist; innovation; interest; multiple omics; nano-string; new therapeutic target; next generation sequencing; novel; novel marker; novel strategies; programs; response biomarker; screening; targeted treatment; transcriptome; transcriptome sequencing; tumor; tumor progression

Biomarkers Core SUMMARY/ABSTRACT The Biomarkers Core provides a centralized resource for rapid, high throughput quantification of transcripts, proteins, and phosphoproteins. Transcripts will be quantified using quantitative reverse transcriptase PCR (RT-qPCR). In addition, the Biomarkers Core will provide genome-wide microarray analysis using an Illumina Beadstation. Next-generation sequencing is available with Illumina sequencers. Quantification of protein levels in blood or tissue lysates will be done using a MesoScale Imager or by reverse-phase protein array (RPPA). NanoString multi-omics will be available for simultaneous RNA/DNA/protein studies. The Biomarker Core will provide: 1) quantitative mRNA levels for known genes that are involved in proliferation and implicated in cancer progression; 2) identification of unique genes and expression profiles in cells or tissues after a molecular or pharmacologic manipulation; 3) validation of the expression of genes that are initially identified in screening by microarrays or RNA-seq; 4) quantification of protein levels both as an independent validation technique and to determine the relationship between transcript levels and protein levels; 5) assessment of changes in protein/phosphoprotein levels using a comprehensive panel of validated antibodies by RPPA. Specific Aims 1-4 represent interactions between SPORE Projects and the Biomarkers Core. Specific Aim 1 Novel Targeted Strategies for Prevention and Conservative Management of Complex Atypical Hyperplasia and Grade 1 Endometrioid Endometrial Cancer. This project will use RNA-seq following microdissection of glands and stromal cells to identify transcripts in CAH/grade 1 endometrioid endometrial cancer that correspond to progestin-responsive versus progestin-resistant disease, as well as markers of response to everolimus. QPCR and proteomics (NanoString multi-omics or RPPA) will be used for validation. Specific Aim 2. CTNNB1 Mutation and Wnt Pathway Activation Define Clinically Aggressive Endometrioid Endometrial Carcinoma. This project will create at least 7 cell lines containing specific mutations and use transcript and miRNA profiles of the stable lines to identify target genes and pathways. QPCR and proteomic analysis will be used to validate candidate transcripts identified in the screening studies. Specific Aim 3 EphA2 Targeting in Uterine Carcinoma. These studies will evaluate the function of EphA2 in cancer and conduct a clinical trial of EPHARNA, a novel approach to deliver siEphA2 systemically using a neutral liposome nanovehicle. Pre- and post-treatment biopsies from patients enrolled in the trial will be analyzed by RPPA and QPCR. Specific Aim 4 A Framework for Identification of Novel Targeted Therapy Combinations in Endometrial Cancer. Aims 1 and 2 of this proposal will use RPPA analysis (~500 samples) to assess treatment efficacy in clinical trials of PARP and PI3K inhibitors (Aim 1) and to map downstream molecular changes for subsequent design of rational drug combinations (Aim 2). RNA-Seq or NanoString multi-omics can also be used to analyze biopsy specimens.

生物标志物核心总结/摘要 生物标志物核心提供了一个集中的资源，用于快速，高通量定量转录本， 蛋白质和磷蛋白。将使用定量逆转录酶PCR对转录物进行定量 （RT-qPCR）。此外，生物标志物核心将提供全基因组微阵列分析， 珠站。下一代测序可通过Illumina测序仪进行。定量蛋白质 将使用MesoScale成像仪或通过反相蛋白质阵列进行血液或组织裂解物中的水平测定 （RPPA）。NanoString多组学将可用于同时进行RNA/DNA/蛋白质研究。生物标志物 核心将提供：1）参与增殖和涉及增殖的已知基因的定量mRNA水平。 在癌症进展中的作用; 2）在癌症治疗后细胞或组织中鉴定独特基因和表达谱， 分子或药理学操作; 3）验证最初在基因组中鉴定的基因的表达， 通过微阵列或RNA-seq进行筛选; 4）定量蛋白质水平，作为独立验证 技术，并确定转录水平和蛋白质水平之间的关系; 5）评估 通过RPPA使用一组全面的经验证的抗体检测蛋白质/磷蛋白水平的变化。 具体目标1-4代表SPORE项目和生物标志物核心之间的相互作用。具体目标1 预防和保守治疗复杂非典型增生的新靶向策略， 子宫内膜样癌1级。该项目将在腺体显微解剖后使用RNA-seq 和基质细胞，以鉴定CAH/1级子宫内膜样癌中的转录物， 孕激素反应性疾病与孕激素抵抗性疾病，以及对依维莫司反应的标志物。qPCR 和蛋白质组学（NanoString多组学或RPPA）将用于验证。具体目标2。CTNNB1 突变和Wnt通路激活定义临床侵袭性子宫内膜样子宫内膜癌这 该项目将创建至少7个含有特定突变的细胞系，并使用转录本和miRNA谱， 稳定的品系来鉴定靶基因和途径。QPCR和蛋白质组学分析将用于验证 筛选研究中确定的候选转录本。特异性目的3 EphA 2在子宫中的靶向 Carcinoma.这些研究将评估EphA 2在癌症中的功能，并进行临床试验。 EPHARNA，一种使用中性脂质体纳米载体全身递送siEphA 2的新方法。前和 将通过RPPA和QPCR分析入选试验的患者的治疗后活检。具体目标4 子宫内膜癌新型靶向治疗组合的鉴定框架。目标1和2 该提案的一部分将使用RPPA分析（约500份样本）来评估PARP临床试验中的治疗疗效 和PI 3 K抑制剂（目标1），并绘制下游分子变化，用于后续合理药物的设计 组合（目标2）。RNA-Seq或NanoString多组学也可用于分析活检标本。