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: 10634587
• 负责人: LEONIDAS G. KONIARIS
• 金额: $ 31.52万
• 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/10634587
• 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; Dedications; Development; Diagnosis; Disease; Ensure; Exhibits; Fibroblasts; Gene Expression; Gene Expression Profiling; Goals; Histologic; Human; Immune system; Indiana; Individual; Infrastructure; Interleukin-6; Knowledge; Macrophage; 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; Specimen; Technology; Testing; Tissue Sample; Tissues; Treatment-related toxicity; Tumor Biology; Tumor Promotion; Universities; Wasting Syndrome; Work; cancer cachexia; clinical translation; effective therapy; human data; human tissue; improved; mortality; mouse model; neoplastic cell; novel; pancreatic cancer patients; preservation; programs; receptor; sample collection; single cell analysis; single nucleus RNA-sequencing; single-cell RNA sequencing; success; transcriptome sequencing; transcriptomic profiling; 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 该计划的目标是增进对胰腺导管腺癌 (PDAC) 的了解，这是一种难治性癌症 恶病质发生率最高的癌症之一。项目 1 重点关注 PDAC 诱导的循环因素，包括 IL-6 及其可溶性受体 (sIL6R)，通过激活 STAT3 和 NF-kB 导致靶组织消耗 存在于脂肪细胞和肌纤维中。项目 2 研究骨骼肌微环境中的 IL-6 和 NF-kB 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的 研究探讨了肿瘤细胞、成纤维细胞和巨噬细胞之间 IL-6、STAT3 和 NF-kB 的相互作用 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问题 它为患者创造。核心 B 将为项目提供前所未有的质疑 PDAC 的潜力 宏观环境，通过提供 1) 人类临床标本来评估其转化潜力 研究结果，以及 2) 使用单细胞和单细胞在单细胞水平上进行转录组分析的能力 PDAC 患者匹配样本中的细胞核 RNAseq，以及 3) 对高级测序的专门支持 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 恶病质。很少有研究报告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，并且没有人尝试将分析与 组织。此外，这些在小鼠 PDAC 宏观环境中以单细胞分辨率进行的开创性研究 模型和人类，由处于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 场地。具体目标是 1) 收集、存储和分发临床注释的 PDAC 恶病质 生物样本。将从印第安纳州的 PDAC 手术患者身上收集临床数据和生物样本 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 General 的肌肉和脂肪样本 患有良性病症的手术患者以及来自 IUSCCC PDAC 手术的肌肉、脂肪和肿瘤样本 患有或不患有中度恶病质的患者将被处理以制备单核制剂。批量 样本将接受 snRNAseq 以测试项目 1-3 中每个项目的具体假设以及总体 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 将为项目 1 的目标 2 中的人类样本的 snRNAseq 数据提供专门的高级分析 3，以及来自项目 1-3 中每个项目的小鼠的 sn 或 scRNAseq 数据。我们将共同确定 PDAC 中的可靶向脆弱性可促进肿瘤杀伤，同时保护宿主组织以保持功能， 改善生活质量和治疗反应，最终提高 PDAC 患者的生存率。