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Mechanisms and optimization of endosomal escape for cell delivery applications

细胞递送应用的内体逃逸机制和优化

基本信息

批准号：
9276732
负责人：
Jean-Philippe Pellois
金额：
$ 27.76万
依托单位：
TEXAS A&M AGRILIFE RESEARCH
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-06-01 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9276732
关键词：
Affect Arginine Biochemical Biological Biological Assay Cells Chemicals Complement Data Development Dimerization Disease Drug Delivery Systems Endocytosis Endosomes Extravasation Foundations Future Future Generations Genes Human Development Hydrophobicity Knowledge Label Lead Learning Libraries Liposomes Masks Mediating Medical Research Membrane Methodology Modeling Molecular Nucleic Acids Organelles Oxidative Stress Patients Penetration Peptides Problem Solving Process Property Proteins RNA RNA Interference Reagent Refractory Research Research Personnel Small Interfering RNA Structure Structure-Activity Relationship System Therapeutic Tumor Suppressor Proteins Variant analog base biophysical analysis cancer cell chemical property cytotoxicity design dimer disulfide bond experimental study human disease improved in vivo knock-down late endosome macromolecule prototype public health relevance repaired therapeutic development therapy development tool trafficking

项目摘要

DESCRIPTION (provided by applicant): Agents that can deliver cell-impermeable biologics inside live cells have the potential to greatly improve the treatment of human diseases and benefit medical research in general. The delivery of peptides, proteins, or siRNAs into cells can for instance be used to reintroduce tumor-suppressors into cancer cells or to knock down disease-causing genes by RNA interference. Yet, methodologies that deliver macromolecules into cells are inefficient and this bottleneck has greatly limited the development of protein or RNA-based therapies. Over the past decade, cell-penetrating peptides (CPPs) have generated a lot of enthusiasm because of their ability to carry macromolecular cargos into cells. A major obstacle to the use of CPPs is that, while they are able to enter cells by endocytosis, CPPs and cargo are retained in endosomes, greatly limiting their usefulness. We have recently uncovered the ability of a specific CPP derivative, a disulfide-bonded dimer of fluorescently labeled TAT (dfTAT), to escape endosomes with astonishingly high efficiency. Consequently, we have further established that dfTAT can deliver proteins into live cells with great ease. Remarkably, the endosomal escape mediated by this agent is not cytotoxic. dfTAT is therefore an extremely promising delivery agents that holds the secret to effective and safe cellular penetration. The objectives of this proposal are to establish the mechanisms of dfTAT-mediated endosomal escape and to identify molecular and cellular features required for this activity. We will identify the structural and molecular determinants of dfTAT endosomal escape by establishing critically needed structure- activity relationships. In addition, we will identify the triggers of endosomal leakage and establish the cellular factors that contribute to this process. The rationale for the proposed research is that the mechanistic knowledge gained will permit the design of improved endosomolytic reagents that can be optimally incorporated into therapeutically relevant drug delivery systems. This should in turn greatly facilitate the development of protein and RNA-based therapeutics and benefit researchers as well as patients.