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We are finding out that we can use the Odyssey CLx system for just about everything protein-related we do in the lab.

Dr. Jeremy Chambers
Florida International University, Miami, FL

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Dr. Jeremy Chambers

At Florida International University in Miami, FL, assistant professor Dr. Jeremy Chambers is harnessing networks of ‘cellular’ powerhouses to monitor drug toxicity. His research group is interpreting the mitochondrial ‘talk’ to design diagnostic assays that can improve chemotherapeutic outcomes in patients.

In cells, mitochondria exist in fused networks that resemble filaments. "When a part of the network is damaged, mitochondria signal the cell to recruit fission machinery. If the damaged mitochondrion is repaired, it rejoins the network. Otherwise, it could lead to an apoptotic response. All of this is brought about by cellular communication involving scaffold proteins," Chambers says.

Chambers studies scaffold proteins housed on the outer mitochondrial membrane that facilitate communication between proteins of signal transduction pathways and the mitochondria. One such scaffold, Sab1 localizes c-jun n-terminal kinase (JNK) on mitochondria and influences the fate of the cell. "Excess of Sab and JNK lead to cytotoxicity and apoptosis," says Chambers. The lab has established a correlation between Sab levels and chemosensitivity in tumor cells. "If Sab level in a cell is low, it will be chemoresistant, whereas higher Sab levels will be more amenable to certain drug treatments."

Statistically-Significant Data with Infrared Fluorescence

The Odyssey CLx System gives us quantitative and reproducible results.

Chambers’ lab is equipped with an Odyssey® CLx Infrared Imaging System that the group uses extensively to perform In-Cell Western™ assays (ICW), quantitative Western blotting analysis, and other applications. "We are finding out that we can use the Odyssey CLx system for just about everything protein-related we do in the lab."

Chambers is profiling various ovarian cancer cell lines for Sab expression levels and drug response using the ICW assay. "ICW assays have enabled the lab to use smaller sample sizes and obtain statistically significant data. It is easy to normalize with Sapphire700™ Stain. The Odyssey CLx System gives us quantitative and reproducible results. We have Z-scores greater than 0.7, which is often seen for some of the best high-throughput assays," he says.

The group also performs On-Cell Western assays for studying the expression of surface molecules that are influenced by mitochondrial pathways. Additionally, they are using the Odyssey CLx System for immunofluorescence imaging of tissue sections. "We have performed brain slice imaging and heart slice imaging on the Odyssey CLx Imager as well. From years of doing immunofluorescence, I can say that it is an advantage to image in the near-infrared (NIR) range where there is very little background fluorescence. We are very excited about the LI-COR® [NIR] technology."

Robust Platform for Quantitative Assay Development

Chambers became acquainted with the Odyssey System during his Post-doc at Scripps Research Institute. "I always promised myself that when I started up my lab that the first instrument I was going to have included in my start up package would be an Odyssey Imaging System. And, that is what I did. From an analytical standpoint, it is good to have an instrument that is quantitative and gives data that is meaningful. Our lab members are fans of the Odyssey [System]."

Chambers and his group are developing their Sab expression assay into a diagnostic platform for ovarian cancer that can predict chemotherapeutic success. "Ovarian cancer is one of the most lethal malignancies in women due to late recognition and recurrence. By focusing on this scaffold, we may be able to polarize mitochondria for cell death, prevent recurrence, and have a more effective treatment at onset." With this diagnostic technology, they hope to be able to test tumor biopsies, determine which drug may not work for an individual patient and thus prevent exposure to toxic drugs.

Cancer researchers like Chambers are developing innovative, personalized diagnostics to improve clinical outcomes with the Odyssey CLx Infrared Imaging technology.

Publications

Publications resulting from work on the Odyssey CLx

  1. Jeremy W. Chambers, Shannon Howard and Philip V. LoGrasso; Neurobiology: Blocking c-Jun N-terminal Kinase (JNK) Translocation to the Mitochondria Prevents 6-Hydroxydopamine-induced Toxicity in Vitro and in Vivo. doi: 10.1074/jbc.M112.421354 originally published online November 26, 2012


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