This project features the integration of advanced photonic technology and microfluidics to attack a major unmet challenge in cell biomechanics. The cellular microenvironment critically regulates cellular function by providing a complex mixture of biochemical and biophysical stimuli. Among the components of the cell-microenvironment interaction, the role of biomechanical factors is recognized to be crucial. In recent years, tremendous progress has been achieved in developing single-cell tools for mechanical stimulation and force response. One area of needed improvement is the non-invasive measurement of intracellular elasticity. Elasticity mediates the transmission of forces inside the cell and the deformation experienced by cell regions under an applied force. However, current technology for cell/ECM elasticity measurements is limited to point-sample analysis or requires contact. These are important limitations since cells are heterogeneous, alter their properties upon mechanical perturbation, and need to be studied in 3D microenvironments. This project will develop an all-optical approach to this unmet need. Brillouin cellular microscopy can map the intracellular elasticity at high resolution, non-perturbatively, without contact in 3D cultures. Brillouin information on cell elasticity will be co-located with fluorescent-based detection of cytoskeletal components and intracellular mechanotransduction. Integration with microfluidic platforms will enable tight control of microenvironment conditions. After instrument validation, I will focus on breast cancer cell migration. Based on preliminary data and literature evidence, I formulated and will test the hypothesis that intracellular elasticity mediates migratio, namely that optimal cell modulus and elasticity polarization are mechanical requirements of the migration machinery and can be used to explain the enhanced motility exhibited by metastatic cells compared to their non-cancerous counterparts. Beyond the cell migration studies, the novel instrumental platform developed and validated during this award, will be broadly applicable as it provides unique quantitative metrics to relate cell- microenvironment mechanical interaction to cell behavior. This K25 award would enable the candidate's transition to research in cellular biomechanics and mechanobiology by complementing his demonstrated expertise in optical technology development with the necessary training in cell biology, microfluidics and ethical research conduct through formal coursework, interaction with mentors and hands-on laboratory training.

Public Health Relevance

This project will develop and validate Brillouin cellular microscopy as a novel optical technology that will uniquely enable non-perturbative and high-resolution measurements of cell-matrix biomechanics in three- dimensional cultures on microfluidic platforms. Integration with fluorescent-based detection of cytoskeletal components and intracellular force transduction will enable studying the role of biomechanical factors in the regulation of cell migration in healthy vs metastatic breast cancer cells. The K25 award will facilitate the candidate's transition from physical sciences and optical technology development to biological research by providing the necessary training in cell biology, biomechanics, microfluidics and ethical research conduct.

Agency
National Institute of Health (NIH)
Type
Mentored Quantitative Research Career Development Award (K25)
Project #
5K25EB015885-02
Application #
8651437
Study Section
Special Emphasis Panel (ZEB1)
Program Officer
Erim, Zeynep
Project Start
Project End
Budget Start
Budget End
Support Year
2
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Massachusetts General Hospital
Department
Type
DUNS #
City
Boston
State
MA
Country
United States
Zip Code
02199
Scarcelli, Giuliano; Besner, Sebastien; Pineda, Roberto et al. (2014) Biomechanical characterization of keratoconus corneas ex vivo with Brillouin microscopy. Invest Ophthalmol Vis Sci 55:4490-5
Kling, Sabine; Akca, Imran B; Chang, Ernest W et al. (2014) Numerical model of optical coherence tomographic vibrography imaging to estimate corneal biomechanical properties. J R Soc Interface 11:20140920