Cancer Studies Using a Novel 3D Printed Zebrafish Intubation Chamber for Longitudinal Imaging

Metabolic plasticity has been shown to be a "hallmark of cancer". Specifically, cancer cells must undergo metabolic evolution to survive and adapt to new organ microenvironments. Recent studies have also shown that the metabolic states of the tumor cells are modulated by the crosstalk between tumor and stromal cells such as endothelial cells. Preclinical models enable recapitulation of physiological environments to understand drug efficacy in distinct microenvironments. We have recently shown that we can employ fluorescence lifetime microscopy to measure longitudinal changes in aerobic glycolysis in 3D culture models and a zebrafish metastatic melanoma model in vivo. We observed different metabolic states for tumors as a function of different organs. Complementary single cell sequencing revealed that there is a difference in immune infiltration associated with the organ specific metastasis. Thus, our goal is to perform long term imaging of transgenic animals where the immune cells are fluorescently labeled. The plan is to introduce these tumor cells at the larval stage so that we can:

a) Perform high throughput confocal imaging of larval fish.
b) Capture ~24 hr continuous imaging of adult fish where tumors are larger than 100 cells.

The first step will help determine how early dynamics and stromal interaction determines the formation of the tumors. The latter will monitor immune infiltration in real time in the presence or absence of applied therapeutics.

The Instrumentation Development and Engineering Application Solutions (IDEAS) has developed a compact zebrafish intubation chamber to enable imaging of a zebrafish with an inverted confocal microscope. The chamber will enable long term imaging of a sedated zebrafish without doing harm to the fish such that they can be reimaged as they grow/mature. The system uses a series of pumps to irrigate warm, aerated water mixed with a sedative into the mouth of the zebrafish and extract the water from the surrounding area while maintaining a constant water level to cover the fish. The system also includes fail-safes to protect the microscope.

This BESIP project will focus on validation and initial experiments using the above setup. The student will work with their NIBIB mentors to evaluate the system, provide feedback on its performance, and assist design changes to improve function and reliability. The student will also work closely with their NCI mentors to perform imaging experiments for longitudinal cancer studies as well as the subsequent analysis. This will be a hands-on, in person project best suited for students with interests in biomedical engineering and cell biology. Working closely with an interdisciplinary team, the student will gain valuable experience with multiple procedures and technologies including animal research, confocal microscope imaging, cancer research, and longitudinal image acquisition.

Tanner lab: The Tanner lab is focused on the early stages of metastasis, when one or a few cells have migrated from the primary tumor and survived transport in blood and lymph vessels to colonize distant organs. In the field of mechanobiology, mechanical properties have been shown to modulate gene expression and phenotype and thus influence cell fate decisions. To fully understand the interplay between the physical cues from the microenvironment and biological processes requires that we resolve and define physical properties of cells, tissues on multiple length scales (sub-microns to millimeters), and temporal scales (milliseconds to days). This is crucial in our efforts to understand such a critical determinant of tumor cell survival that can last for many years in multiple organs. Deciphering these physical cues encountered along the metastatic cascade in vivo presents a conundrum.  The Tanner lab focused on this gap by adopting and modifying intravital microscopy and optical tweezers to address the hypothesis that tissue biophysics regulates organ selectivity and tumor outgrowth during metastasis in vivo.

Pohida lab: Provides electrical, electronic, electro-optical, mechanical, computer, and software engineering expertise to NIH projects that require in-house technology development. Collaborations involve advanced signal transduction and data acquisition; real-time signal and image processing; control and monitoring systems (e.g., robotics and process automation); and rapid prototype development. Collaborations result in the design of first-of-a-kind biomedical/clinical research systems, instrumentation, and methodologies.