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Multiplex single-cell analysis of intracellular protein signaling dynamics

细胞内蛋白质信号传导动力学的多重单细胞分析

基本信息

批准号：
10261852
负责人：
Min Xue
金额：
$ 6.45万
依托单位：
UNIVERSITY OF CALIFORNIA RIVERSIDE
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-07 至 2022-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10261852
关键词：
Adopted Affect Affinity BRAF gene Bar Codes Biological Biological Assay Biological Process Biomedical Research Cell Line Cells Characteristics Clinical Complex Data Set Detection Development Disease Progression Disease Resistance Eligibility Determination Epitopes Event Exercise Exhibits Fluorescent Probes Genetic Goals Heterogeneity Image Imaging Techniques Immobilization Immune response Institution Intracellular Signaling Proteins Libraries MAPK8 gene MCF7 cell MEKs Mass Spectrum Analysis Measurement Metabolic Metabolism Methods Microscope Modification Monitor Nature Oncology Peptide Library Peptides Pharmaceutical Preparations Phosphorylation Process Protein Analysis Protein Dynamics Proteins Proteomics Proto-Oncogene Proteins c-akt Protocols documentation Research Personnel Resistance development Resolution Sampling Scanning Signal Transduction Signaling Protein Speed Statistical Data Interpretation Surface System Technology Therapeutic Time Treatment Efficacy Uncertainty Work analytical method base biological systems cell type design experience fluorophore improved inhibitor/antagonist insight non-genetic protein expression prototype screening single cell analysis single cell proteins single cell technology success therapeutic evaluation tumor progression

项目摘要

ABSTRACT Recent developments on single-cell analytical methods have provided a clarifying view on cellular heterogeneity and its associated mechanistic and therapeutic implications. Nevertheless, single-cell studies on intracellular protein signaling activities can be confounded by the dynamic nature of signaling events. In addition, there is a pressing need of developing non-genetic analytical methods for clinical samples. These requisites call for a single-cell approach that can interrogate intracellular protein signaling dynamics while being compatible with other downstream analytical methods. A recently developed prototype method has enabled dynamic probing of intracellular protein signaling activities at single-cell resolution, without genetic modifications. The technology was based on intracellularly delivered epitope-targeting peptide probes, a microwell-based single-cell chip and high-speed imaging techniques. However, many obstacles exist that challenge the generalizability of this approach and limit its biomedical application. In this proposal, the aim is to significantly advance this proof-of- concept technology. Five specific goals are included: develop an automated screening protocol for improved screening efficiencies, implement a unique bicyclic peptide library for higher biding affinities, employ cell permeabilizers as a more generalizable delivery method, achieve simultaneous analysis of multiple signaling proteins, and integrate this technology with other single-cell detection methods. This integrated multiplex pipeline will provide an enabling technology that promises a more in-depth understanding of the interplay among protein signaling activities, protein expression levels and metabolism in biological processes.

摘要