Microfluidic-Enabled Intracellular Delivery of Membrane Impermeable Inhibitors to Study Target Engagement in Human Primary Cells
| Autor: | Brian Juba, Betsy Pierce, Steve Kortum, Armon Sharei, Jing Li, Erik C. Hett, Jean-Baptiste Telliez, Bu Wang, Mark E. Bunnage, Lee R. Roberts, Strohbach Joseph Walter, Michael Pacheco, Atli Thorarensen, Lyn H. Jones, Michael L. Vazquez, Jonathan B. Gilbert |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Membrane permeability 02 engineering and technology Biology Biochemistry Cell membrane Structure-Activity Relationship 03 medical and health sciences Lab-On-A-Chip Devices medicine Humans Janus Kinase Inhibitors Structure–activity relationship Cells Cultured Janus Kinases Molecular Structure Electroporation Cell Membrane General Medicine Biochemical Activity 021001 nanoscience & nanotechnology Small molecule Cell biology 030104 developmental biology medicine.anatomical_structure Leukocytes Mononuclear Molecular Medicine 0210 nano-technology Janus kinase Intracellular |
| Zdroj: | ACS Chemical Biology. 12:2970-2974 |
| ISSN: | 1554-8937 1554-8929 |
| Popis: | Biochemical screening is a major source of lead generation for novel targets. However, during the process of small molecule lead optimization, compounds with excellent biochemical activity may show poor cellular potency, making structure-activity relationships difficult to decipher. This may be due to low membrane permeability of the molecule, resulting in insufficient intracellular drug concentration. The Cell Squeeze platform increases permeability regardless of compound structure by mechanically disrupting the membrane, which can overcome permeability limitations and bridge the gap between biochemical and cellular studies. In this study, we show that poorly permeable Janus kinase (JAK) inhibitors are delivered into primary cells using Cell Squeeze, inhibiting up to 90% of the JAK pathway, while incubation of JAK inhibitors with or without electroporation had no significant effect. We believe this robust intracellular delivery approach could enable more effective lead optimization and deepen our understanding of target engagement by small molecules and functional probes. |
| Databáze: | OpenAIRE |
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