INTEGRATED MICRO SCALE TRANSCRIPTIONAL PROFILING OF CELL COMMUNICATION NETWORKS


Year of Award:
2011
Award Type:
R33
Project Number:
CA160344
RFA Number:
RFA-CA-10-004
Technology Track:
Molecular & Cellular Analysis Technologies
PI/Project Leader:
BEEBE, DAVID J
Other PI or Project Leader:
ALARID, ELAINE T
Institution:
UNIVERSITY OF WISCONSIN-MADISON
In cancer, differences in risk factors and genetics generate diverse clinical behaviors (disease progression, response to therapy) at the patient level. Yet, despite critical implications for therapeutic outcome, few tools are available to study patient samples at the cellular level in a rigorous way with sufficient replicates to enable clinically meaningful assessments. This challenge is particularly acute in the context of cancer metastasis, where micro-tumors establish themselves in ectopic environments which are significantly different in cellular make-up than primary tumor. But few tools allow one to probe intercell network communication (e.g. paracrine signals between different cell types). A sample preparation bottleneck is particularly acute for the study of heterogeneity requiring large numbers of samples (e.g. each patient sample is different, each requiring nucleic acid purification). Thus, there is a need to perform culture (including co-culture), treatment, and analyte purification from a large number of samples (multiple patients, treatments, and repeats). Our goal is to apply simple micro scale technology to profile intercell regulation on heterogeneous patient samples. The cornerstones of our approach are an immiscible oil barrier, termed the 'Phase-Gate,' that separates the upstream 'dirty' side (culture, treatment, lysis) of sample processing from the downstream 'clean' side (analyte purification and detection) and micro chamber co-culture with increased sensitivity to paracrine signaling. Our preliminary data predicts that neighboring stroma play a critical role in regulating key therapeutic targets and that advances in experimental capabilities (throughput, sensitivity, compartmentalization) will provide new tools and biological insights that will advance basic and translational cancer research. Here, we will apply a co-culture and transcriptional analysis platform to dissecting cell network communication - specifically how intercell regulation contributes to nuclear receptor regulation and ultimately to tumor progression and therapeutic resistance. The validation and advanced development of this technology directed towards an important area in cancer biology (nuclear receptor response) will accelerate its use enable studies not feasible previously. PUBLIC HEALTH RELEVANCE: We anticipate that the research proposed in this grant will not only help to elucidate how intercell regulation contributes to nuclear receptor regulation, but ultimately to tumor progression and therapeutic resistance. More broadly these studies will provide new tools and biological insights that will advance basic and translational cancer research.