? The emergence of effective therapies for modulating disordered signal transduction in human disease has brought about a crucial need to assay the activity of signaling proteins in patients and model organisms. To meet this need, the current grant application proposes to design, build, and test an analytical platform for the performance of single-cell kinase assays on primary patient cells. State-of-the art microfabrication technology will be employed to engineer a microfluidic system that integrates electronics and photonics for the performance of microanalytical chemical separations. The microfluidic system will be coupled to a meso-scale incubator for sample preparation. The components of the hybrid device will be designed and optimized individually for their given function, and then integrated into a multifunctional platform. This platform will be used to carry out a newly developed method for assaying the catalytic activity of specific kinases in individual cells. In the final phase of the work assays of kinases known to be essential for the development and maintenance of an exemplary disease will be demonstrated in patient samples. The activity of the tyrosine kinase Bcr-Abl and the serine/threonine kinase protein kinase B in the cells of patients with chronic myelogenous leukemia will be assessed after exposure to varying doses of the recently approved kinase inhibitor imatinib mesylate. The direct assay of kinases before and after ex vivo exposure of the patient's cells to this pharmaceutical compound will provide unique and clinically valuable data regarding the drug's efficacy and dosing for personalized tailoring of treatment options. The single-cell data will also enable subpopulations of cells within a given patient to be assessed to address cell-to-cell heterogeneity in response to therapy. A number of kinase inhibitors are now in development or clinical trials for a broad range of diseases making the development of this novel instrumentation of widespread value for biomedical research and clinical application. ? ? ?
| Tsai, Becky Pinjou; Jimenez, Judith; Lim, Sharon et al. (2014) A novel Bcr-Abl-mTOR-eIF4A axis regulates IRES-mediated translation of LEF-1. Open Biol 4:140180 |
| Phillips, K Scott; Lai, Hsuan Hong; Johnson, Emily et al. (2011) Continuous analysis of dye-loaded, single cells on a microfluidic chip. Lab Chip 11:1333-41 |
| Hellman, Amy N; Vahidi, Behrad; Kim, Hyung Joon et al. (2010) Examination of axonal injury and regeneration in micropatterned neuronal culture using pulsed laser microbeam dissection. Lab Chip 10:2083-92 |
| Wang, Yuli; Dhopeshwarkar, Rahul; Najdi, Rani et al. (2010) Microdevice to capture colon crypts for in vitro studies. Lab Chip 10:1596-603 |
| Phillips, K Scott; Kang, Kyung Mo; Licata, Louise et al. (2010) Air-stable supported membranes for single-cell cytometry on PDMS microchips. Lab Chip 10:864-70 |
| Xu, Wei; Wu, Liang L; Zhang, Yang et al. (2009) A Vapor Based Microfluidic Flow Regulator. Sens Actuators B Chem 142:355-361 |
| Borland, Laura M; Kottegoda, Sumith; Phillips, K Scott et al. (2008) Chemical analysis of single cells. Annu Rev Anal Chem (Palo Alto Calif) 1:191-227 |
| Phillips, K Scott; Kottegoda, Sumith; Kang, Kyung Mo et al. (2008) Separations in poly(dimethylsiloxane) microchips coated with supported bilayer membranes. Anal Chem 80:9756-62 |
| Quinto-Su, P A; Venugopalan, V; Ohl, C-D (2008) Generation of laser-induced cavitation bubbles with a digital hologram. Opt Express 16:18964-9 |
| Lai, Hsuan-Hong; Quinto-Su, Pedro A; Sims, Christopher E et al. (2008) Characterization and use of laser-based lysis for cell analysis on-chip. J R Soc Interface 5 Suppl 2:S113-21 |
Showing the most recent 10 out of 28 publications
