Investigator: Paul Campagnola University of Wisconsin
Campagnola laboratory is a leader in developing bioimaging techniques to study the role of extracellular matrix (ECM) in cancer as well as in connective tissue disorders. To this end, his lab uses Second Harmonic Generation (SHG) microscopy for imaging structural aspects of tissues. He further pioneered microfabrication techniques for creating biomimetic cell culture platforms that recapitulate the native 3D tissue ECM environment to study signaling pathways associated with cancer and also to provide insight into the design of tissue engineering scaffolds.
The extracellular matrix (ECM) directs cell shape, spreading, differentiation, migration, and proliferation in addition to providing structural support [1] by presenting a complex milieu of topographic, mechanical and biochemical cues to cells. For example, adhesion and migration are known to become mis-regulated during ovarian and other cancers [2]. It is recognized that the response of a patient to a particular treatment or a particular drug will be very different from that of another person due to the unique genotype and phenotype. In drug selection for disease treatment, tissue mimics created from a patient's own cells growing on ECM mimics that recreate the patient's tissue morphology may allow clinicians to select the most effective drug on a rational basis instead of today's trial-and-error approach. As an example, for ovarian cancer treatment, one may envision cancer and stromal cells extracted during biopsy may be expanded, and patterned into tissue mimics in a fully humanized environment. These tissue mimics may be automatically cultured in microfluidic devices until they become morphologically "mature" lesions. A drugs panel at different dosages can be used to challenge these tissue mimics and to identify a precise dosing scheme (Fig. 1).
The native ECM has intrinsic 3D complexity with size features over length scales of a ~100 nm in diameter to several µm in length. The fabrication of these tissue mimics, to the fullest extent possible, needs to faithfully reproduce the nano/microstructured topography from the same multiple ECM components that comprise the in vivo structure. Recent work in the Campagnola lab, a leader in developing bioimaging techniques to study the role of extracellular matrix (ECM) in pathogenesis, utilizes multiphoton microfabrication technology to create a biomimetic environment following a "blueprint" based on high resolution microscopy imaging data [3,4] (Fig. 2).
Campagnola lab has also demonstrated that cells grown in these biomimetic environments exhibit more realistic proliferation rates and migration dynamics (speed and directionality as shown in Fig. 3) as compared with less native environments. Working with ovarian cancer expert, Dr. Patankar [5], they further showed the normal and ovarian cancer cell phenotype can be modified just by varying their scaffold morphology.
LBRC will adapt Campagnola's microfabrication approach and will attempt to increase fabrication speed by at least two orders of magnitude using wide-field temporal focusing approach extending a well proven technique in our laboratory. The increased fabrication speed will be obtained from parallelization of excitation beam. The benchmark for transferring the point scanning technology to wide-field system will be 90% fidelity verified by 3D imaging. Working with Campagnola, we will use these scaffolds to establish a drug assay protocol and demonstrate differential response of ovarian cancer cells to cisplatin, oxaliplatin, arsenite, or taxol and combinations thereof under different stromal compositions and morphologies.
Dr. Campagnola (U. of Wisconsin) has pioneered microfabrication techniques for creating biomimetic cell culture platforms that recapitulate the native 3D tissue ECM environment. LBRC will help increase the fabrication speed by incorporating state-of-the-art wide-field tmporal focusing approach developed at LBRC. The developed scaffolds will be further used to study the response of ovarian cancer cells to different drugs.