Core B – Human Biospecimen and Advanced Sequencing Core

核心 B — 人类生物样本和高级测序核心

• 批准号: 10441216
• 负责人: LEONIDAS G. KONIARIS
• 金额: $ 31.64万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10441216
• 关键词: Address; Adipocytes; Adipose tissue; Automobile Driving; Benign; Biometry; Blood; Body Weight decreased; Cachexia; Cell Nucleus; Cell Separation; Cells; Clinical; Clinical Data; Comprehensive Cancer Center; Consent; DNA Sequencing Facility; Data; Development; Diagnosis; Disease; Ensure; Exhibits; Fibroblasts; Gene Expression; Gene Expression Profiling; Goals; Histologic; Human; Immune system; Indiana; Individual; Infrastructure; Interleukin-6; Knowledge; Malignant Neoplasms; Malignant neoplasm of pancreas; Metabolic; Molecular; Morbidity - disease rate; Mus; Muscle; Muscular Atrophy; NF-kappa B; Operative Surgical Procedures; Pancreatic Ductal Adenocarcinoma; Pathology; Patients; Peripheral; Phenotype; Preparation; Process; Protocols documentation; Quality of life; Reporting; Research; Research Personnel; Resolution; Resources; Role; STAT3 gene; Sampling; Sequence Analysis; Signal Transduction; Skeletal Muscle; Small Nuclear RNA; Specimen; Technology; Testing; Tissue Sample; Tissues; Treatment-related toxicity; Tumor Biology; Universities; Wasting Syndrome; Work; cancer cachexia; clinical translation; effective therapy; human data; human tissue; improved; macrophage; mortality; mouse model; neoplastic cell; novel; pancreatic cancer patients; preservation; programs; receptor; sample collection; single cell analysis; single-cell RNA sequencing; success; transcriptome; transcriptome sequencing; translational potential; treatment response; tumor; tumor microenvironment; wasting

PROJECT SUMMARY: CORE B The goal of this Program is to advance knowledge of pancreatic ductal adenocarcinoma (PDAC), a recalcitrant cancer with one of the highest rates of cachexia. Project 1 focuses on PDAC-induced circulating factors, including IL-6 and its soluble receptor (sIL6R), leading to wasting of the target tissues via activation of STAT3 and NF-kB in adipocytes and myofibers. Project 2 investigates IL-6 and NF-kB in the skeletal muscle microenvironment in PDAC cachexia, as well as NF-kB in the tumor microenvironment and its effects on macrophages. Project 3’s studies probe interactions of IL-6, STAT3, and NF-kB among tumor cells, fibroblasts, and macrophages in the tumor microenvironment. Together these studies will tackle both PDAC tumor biology and the metabolic havoc it creates for the patient. Core B will provide the Projects with unprecedented potential to interrogate the PDAC macroenvironment by providing 1) human clinical specimens to evaluate the translational potential of their findings, and 2) the capacity to carry out transcriptome profiling at the single cell level using single cell and single nucleus RNAseq in matched samples from patients with PDAC, and 3) dedicated support for advanced sequence analysis. In doing so, this Core addresses barriers that have historically limited research in PDAC and cancer cachexia. Very few studies have reported gene expression or histological endpoints in peripheral tissues in human cancer cachexia and even fewer in PDAC cachexia, and none have attempted to match analyses across tissues. As well, these pioneering studies at single cell resolution in the PDAC macroenvironment of mouse models and humans, carried out by investigators at the forefront of these technologies and analytical capabilities will enable the Projects to make groundbreaking discoveries while contributing novel, high resolution data to the field. The specific aims are to 1) Collect, store and distribute clinically annotated PDAC cachexia biospecimens. Clinical data and biospecimens will be collected from PDAC surgical patients at Indiana University Simon Comprehensive Cancer Center (IUSCCC) and from patients from the Danish PACTO Trial for centralized compliance, processing, and distribution. 2) Carry out a single nucleus (sn) RNAseq study of the human PDAC cachexia macroenvironment for all Projects. Muscle and adipose samples from IUSM general surgery patients with benign conditions and muscle, adipose and tumor samples from IUSCCC PDAC surgery patients with and without moderate cachexia will be processed to make single nuclei preparations. Batched samples will be subjected to snRNAseq to test specific hypotheses for each of Projects 1-3 and the overall program. 3) Provide advanced analysis of single cell (sc) and snRNAseq studies for all Projects. Core B will provide dedicated advanced analysis of the snRNAseq data from human specimens in Aim 2 for Projects 1- 3, and of sn or scRNAseq data from mice emanating from each of the Projects 1-3. Together we will identify targetable vulnerabilities in PDAC to promote tumor killing while sparing host tissues to preserve function, improve quality of life and response to therapy, and ultimately increase survival for patients with PDAC.

项目摘要：核心B 该程序的目的是提高胰腺导管腺癌（PDAC）的知识 癌症是恶病质率最高的癌症之一。项目1的重点是PDAC引起的循环因素，包括 IL-6及其可溶受体（SIL6R），导致通过Stat3和NF-KB的激活浪费目标时间 在脂肪细胞和肌纤维中。项目2在骨骼肌微环境中研究IL-6和NF-KB 肿瘤微环境及其对巨噬细胞的影响，PDAC Cachexia以及NF-KB。项目3 研究肿瘤细胞，成纤维细胞和巨噬细胞中IL-6，STAT3和NF-KB的探针相互作用 肿瘤微环境。这些研究将共同​​解决PDAC肿瘤生物学和代谢破坏 它为患者创建。核心B将为项目提供质疑PDAC的前所未有的潜力 通过提供1）人类临床标本来评估其翻译潜力 发现，以及2）使用单个单元和单个单元格在单细胞水平上进行转录组分析的能力 来自PDAC患者匹配的样品和3）专用于晚期序列的核心核酸核酸核。 分析。在此过程中，该核心解决了历史上限制PDAC和癌症研究的障碍 卡希克西亚。很少有研究报告了外周组织中基因表达或组织学终点 人类癌症恶病质，在PDAC病原质中甚至更少，没有人试图匹配各种分析 组织。同样，这些在小鼠PDAC宏观环境中单细胞分辨率的开创性研究 模型和人类，由研究人员在这些技术和分析能力的最前沿进行 将使这些项目能够在为小说，高分辨率数据提供针对的同时进行开创性的发现 场地。具体目的是1）收集，存储和分发临床注释的PDAC Cachexia 生物测量。将从印第安纳州的PDAC手术患者中收集临床数据和生物测量 西蒙大学综合癌症中心（IUSCCC）和来自丹麦帕克托试验的患者 集中式合规，处理和分配。 2）进行单个核（SN）RNASEQ研究 所有项目的人类PDAC Cachexia宏观环境。 IUSM一般的肌肉和脂肪样品 具有良性条件和肌肉，脂肪和肿瘤样本的手术患者来自IUSCCC PDAC手术 有或没有中度恶病质的患者将进行处理以进行单核制剂。批处理 样本将接受SNRNASEQ，以测试每个项目1-3的特定假设和整体 程序。 3）为所有项目提供对单细胞（SC）和SNRNASEQ研究的高级分析。核心b 将在AIM 2中对SNRNASEQ数据的SNRNASEQ数据进行专门的高级分析。 3，以及来自每个项目1-3的小鼠的SN或SCRNASEQ数据。我们一起确定 PDAC中有针对性的脆弱性促进肿瘤杀死，同时保留宿主组织以保持功能， 提高生活质量和对治疗的反应，并最终增加PDAC患者的生存率。