High-throughput micro-RNA profiling of single cells and its application in leukemia

单细胞高通量micro-RNA分析及其在白血病中的应用

• 批准号: 10424916
• 负责人: Suman Bose
• 金额: $ 8.36万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-15 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10424916
• 关键词: Address; Bar Codes; Biochemical; Biological; Biological Markers; Biological Process; Biology; Biomedical Engineering; Cell Line; Cell physiology; Cells; Clinical; Cytogenetics; Data; Detection; Development; Device Designs; Devices; Diabetes Mellitus; Diagnosis; Diagnostic; Disease; Disease Pathway; Disease Progression; Drug resistance; Early Diagnosis; Emulsions; Engineering; Ensure; Environment; Family; Gene Expression; Genes; Genetic Transcription; Goals; Heterogeneity; Immune System Diseases; Individual; Institutes; Leukemic Cell; Ligase; Ligation; Light; Malignant Neoplasms; Maps; Measurement; Medical; Medicine; Mentors; Messenger RNA; MicroRNAs; Microfluidics; Modality; Molecular Biology; Nucleic Acids; Oncology; Pathogenesis; Patients; Pattern; Phase; Population; Positioning Attribute; Process; Proteomics; RNA; Reaction; Regulator Genes; Relapse; Research; Resistance; Rest; Sampling; Science; Scientist; Sensitivity and Specificity; Signal Pathway; Structure; Techniques; Technology; Testing; Training; Training Programs; Transcript; Variant; Work; base; career; cell type; detection sensitivity; disease diagnosis; disease mechanisms study; disease phenotype; leukemia; magnetic beads; meetings; miRNA expression profiling; novel therapeutics; outreach; patient population; prognostic value; single-cell RNA sequencing; student mentoring; tool; transcription factor

Project Summary Single-cell RNA sequencing (scRNA-seq) holds the promise to revolutionize the study of biology by allowing measurement of cellular processes within individual cells with unprecedented precision and breadth. Micro-RNAs (miRNA) are a class of small regulatory RNAs that are involved in a range of biological processes such as organismal development, cellular differentiation, and major signaling pathways. miRNA signatures correlate strongly with disease phenotype and can serve as reliable biomarkers. This proposal will develop a microfluidic platform that can allow isolation and sequencing of miRNA from individual single-cells in a highly-parallel and unbiased manner. In the K99 phase, I will sequentially develop each segment of the device and optimize the reactions and processes associated with it. The device will be tested and validated using control cell lines to determine the sensitivity of detection, and establish the quality of single cell data. Finally, I will use the platform to profile miRNA expressions in leukemia cell lines and those obtained from patients with the goal to identify expression patterns that will have diagnostic or prognostic value. The R00 phase of the project will focus on applying this platform in studying disease pathogeneses in leukemia. Findings from the study will shine light on new pathways for disease progression and drug resistance, which can be used for developing new therapeutic modalities. I will also develop the platform as an ultra-sensitive diagnostic tool for detecting leukemia in patients. I also propose an extensive training program that will help me realize my career goals and support my transition to an independent research position. The research environment at Koch Institute is unparalleled and offers the unique opportunity to interact and collaborate with eminent biologists and engineers. I have assembled an exceptional team: Prof. Dan Anderson and Prof. Robert Langer, experts in bioengineering and high-throughput techniques, will mentor me. I will work very closely with Prof. Phillip Sharp, RNA Biologist, who will help me address the molecular biology component of the project. Dr. Amir Faithi will provide clinical samples and direction to increase the medical impact of the project. My training will also involve mentoring students, presenting my work in single-cell meetings, and science outreach.

项目概要 单细胞 RNA 测序 (scRNA-seq) 有望彻底改变生物学研究 通过允许以前所未有的方式测量单个细胞内的细胞过程 精度和广度。 Micro-RNA (miRNA) 是一类小调控 RNA， 参与一系列生物过程，如有机体发育、细胞 分化和主要信号通路。 miRNA 特征与疾病密切相关 表型并可以作为可靠的生物标志物。 该提案将开发一个微流体平台，可以允许分离和测序 以高度平行且公正的方式从单个单细胞中获取 miRNA。在K99阶段，我 将依次开发装置的各个部分并优化反应和工艺 与之相关。该设备将使用对照细胞系进行测试和验证，以确定 检测的灵敏度，并确定单细胞数据的质量。最后我会使用这个平台 分析白血病细胞系和从具有目标的患者身上获得的 miRNA 表达 识别具有诊断或预后价值的表达模式。 该项目的R00阶段将重点将该平台应用于疾病研究 白血病的发病机制。研究结果将揭示疾病的新途径 进展和耐药性，可用于开发新的治疗方式。我 还将开发该平台作为超灵敏诊断工具，用于检测白血病 患者。 我还提出了一个广泛的培训计划，这将帮助我实现我的职业目标和支持 我向独立研究职位的转变。科赫研究所的研究环境是 无与伦比，提供与杰出生物学家互动和合作的独特机会 和工程师。我组建了一支出色的团队：丹·安德森教授和罗伯特教授 兰格是生物工程和高通量技术方面的专家，他将指导我。我会很努力工作 与 RNA 生物学家 Phillip Sharp 教授密切合作，他将帮助我解决分子生物学问题 项目的组成部分。 Amir Faithi 博士将提供临床样本和指导以增加 该项目的医疗影响。我的培训还将包括指导学生、展示我的 从事单细胞会议和科学推广工作。

项目成果

Chemical Tuning of Fibers Drawn from Extensible Hyaluronic Acid Networks.

• DOI: 10.1021/jacs.0c09691
• 发表时间: 2020-11-18
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Chu, Crystal K.;Joseph, Alby J.;Limjoco, Matthew D.;Yang, Jiawei;Bose, Suman;Thapa, Lavanya S.;Langer, Robert;Anderson, Daniel G.
• 通讯作者: Anderson, Daniel G.