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: 10172473
• 负责人: LEONIDAS G. KONIARIS
• 金额: $ 26.66万
• 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/10172473
• 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; response; sample collection; single cell analysis; single-cell RNA sequencing; success; transcriptome; transcriptome sequencing; 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.

项目总结：核心B 本项目的目的是提高对胰腺导管腺癌（PDAC）的认识， 恶病质发病率最高的癌症项目1关注PDAC诱导的循环因子，包括 IL-6及其可溶性受体（sIL-6 R），通过激活STAT 3和NF-κ B导致靶组织消耗 在脂肪细胞和肌纤维中。项目2研究IL-6和NF-kB在骨骼肌微环境中的作用， PDAC恶病质，以及肿瘤微环境中的NF-kB及其对巨噬细胞的影响。项目3 研究了IL-6、STAT 3和NF-kB在肿瘤细胞、成纤维细胞和巨噬细胞中的探针相互作用。 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 为病人创造的。核心B将为项目提供前所未有的潜力，以询问PDAC 通过提供1）人类临床标本来评估其翻译潜力， 发现，和2）在单细胞水平上使用单细胞和单克隆抗体进行转录组分析的能力。 PDAC患者的匹配样本中的核RNAseq，以及3）专用于高级序列支持 分析.通过这样做，该核心解决了历史上限制PDAC和癌症研究的障碍 恶病质很少有研究报道了外周组织中的基因表达或组织学终点， 人类癌症恶病质，甚至更少的PDAC恶病质，没有人试图匹配分析， 组织中同样，这些在小鼠PDAC宏环境中的单细胞分辨率的开创性研究， 模型和人类，由这些技术和分析能力最前沿的研究人员进行 将使这些项目能够取得突破性的发现，同时为全球气候变化提供新颖的高分辨率数据。 领域具体目标是：1）收集、储存和分发临床注释的PDAC恶病质 生物标本将从印第安纳州的PDAC手术患者中收集临床数据和生物标本 西蒙大学综合癌症中心（IUSCCC）和来自丹麦PACTO试验的患者， 集中合规、处理和分发。2)进行单核（sn）RNAseq研究， 所有项目的人类PDAC恶病质宏观环境。来自IUSM一般的肌肉和脂肪样本 患有良性疾病的手术患者以及来自IUSCCC PDAC手术的肌肉、脂肪和肿瘤样本 将处理具有和不具有中度恶病质的患者以制备单个核制剂。批处理 将对样本进行snRNAseq，以测试项目1-3中每个项目的特定假设， 程序. 3)为所有项目提供单细胞（sc）和snRNAseq研究的高级分析。核心B 将为Aim 2中的项目1提供来自人类标本的snRNAseq数据的专用高级分析- 3，以及来自项目1-3中的每一个的小鼠的sn或scRNAseq数据。我们将共同确定 PDAC中的靶向弱点以促进肿瘤杀伤，同时保留宿主组织以保留功能， 改善生活质量和对治疗的反应，并最终提高PDAC患者的生存率。