Using Tumor-Targeting Mutant Salmonella To Treat Prostate Cancer

Analytik-Jena’s iBox® Scientia small animal imaging system (Figure 1) has been specifically developed for imaging fluorescent proteins. The system offers a number of benefits for imaging whole animal models. The availability of fluorescent proteins with a broad range of emission wavelengths simplifies multiple label studies.

Since the proteins are fluorescent by nature, there is no need to use cofactors or substrates. Furthermore, the fluorescence properties of the reporter proteins are continuously being expanded and a range of very bright red emitting proteins are now available. Academic and commercial sources can supply the genes for these reporter proteins.

iBox Scientia small animal imaging system

Figure 1. iBox Scientia small animal imaging system

Expressed Fluorescent Proteins

When combined with Analytik-Jena’s f1.2 optics, the imaging system can provide very rapid exposures to produce sharp images that are not affected by the animal’s breathing. The bright fluorescence enables tumor margins to be easily defined with the and Analytik-Jena’s VisionWorks software area density tools and to make quantitation of the tumor area easily achievable.

Analytik-Jena’s iBox® Scientia small animal imaging system was used to image RFP orthotopic tumors in nude mice over a two-week period (Figure 2).

RFP orthotopic tumors in nude mice imaged over a period of two weeks using Analytik-Jena iBox® Scientia small animal imaging system.

Figure 2. RFP orthotopic tumors in nude mice imaged over a period of two weeks using Analytik-Jena iBox® Scientia small animal imaging system.

As detailed by Zhao et al., the treatment involves injecting a Salmonella typhimurium mutant on a daily basis. This mutant concentrates in tumors and minimizes or eliminates metastasis with PC-3 human prostate tumor cells. This study used U87 red fluorescent protein (RFP) expressing the human glioma cell line.

Materials and Methods

Analytik-Jena’s iBox Scientia in vivo imaging system was configured with the following features:

  • Motorized sample platen
  • CFP, YFP, RFP and GFP emission and excitation filter sets
  • 4.2mpx high resolution BioChemi 500 camera
  • Rapid f1.2 50mm large area Biolens
  • VisionWorks®LS analysis software
  • Automated BioLite™ excitation light source
  • Analytik-Jena heater Warming plate set to 370C

The imaging procedure is straightforward. A small amount of U87 cells were subcutaneously injected into the nude mice. After the tumor had reached 1mm2 in volume, observations were made and designated as Day 0. Weekly injections of 5x107/100µl Salmonella A1-R were received by the treatment group via tail injection.

The RFP excitation and emission filter set was then used to capture the images with Analytik-Jena’s iBox small animal imaging system. Fluorescent and white light images were taken and combined using the blend control (a paste special with destination and source at 25% and 75%, respectively), with the fluorescent image colored according to the fluorescence color, which is red for RFP in this case. The typical exposure times for white light and RFP were 0.010s and 1s, respectively.

Results

A comparison of the untreated control and treatment on days 7 and 14 suggested that the weekly injections of Salmonella A1-R administered intravenously had targeted the tumor and significantly reduced tumor growth.

Conclusion

In this study, the fluorescent imaging of tumors demonstrates how critical the process is in time course studies and how important fluorescent protein imaging is in cancer research studies. The use of Analytik-Jena’s iBox® Scientia small animal imaging system together with Analytik-Jena f1.2 optics yields sharp images that are not affected by an animal’s breathing.

References

  • Hoffman, R.M. and Yang, M. Whole-body imaging with fluorescent proteins. Nat. Protoc. 2006, 1, 1429-1438.
  • Hoffman, R.M. and Yang, M. Color-coded fluorescence imaging of tumor-host interactions. Nat. Protoc. 2006, 1, 928-935.
  • Zhao, M.; Geller, J.; Ma, H.; Yang, M.; Penman, S.; Hoffman, R.M. Monotherapy with a tumor-targeting mutant of Salmonella typhimurium cures orthotopic metastatic mouse models of human prostate cancer. Proc. Natl. Acad. Sci. U. S. A. 2007, 104, 10170-10174.
  • Zhao, M.; Yang, M.; Ma, H.; Li, X.; Tan, X.; Li, S.; Yang, Z.; Hoffman, R.M. Targeted therapy with a Salmonella typhimurium leucine-arginine auxotroph cures orthotopic human breast tumors in nude mice. Cancer Res. 2006, 66, 7647-7652.

About Analytik Jena US

Analytik Jena is a provider of instruments and products in the areas of analytical measuring technology and life science. Its portfolio includes the most modern analytical technology and complete systems for bioanalytical applications in the life science area.

Comprehensive laboratory software management and information systems (LIMS), service offerings, as well as device-specific consumables and disposables, such as reagents or plastic articles, complete the Group’s extensive range of products.

About Life Science

The Life Science product area demonstrates the biotechnological competence of Analytik Jena AG. We provide a wide product spectrum for automated total, as well as individual solutions for molecular diagnostics. Our products are focused to offer you a quality and the reproducibility of your laboratory results.

This will surely ease your daily work and speed up your work processes in a certain way.
All together we support you through the complete process of the lab work. Besides we offer customized solutions and are able to adapt our products to your needs. Automated high-throughput screening systems for the pharmaceutical sector are also part of this segment’s extensive portfolio.

About Analytical Instrumentation

Analytik Jena has a long tradition in developing high-performance precision analytical systems which dates back to the inventions made by Ernst Abbe and Carl Zeiss. We have grown to become one of the most innovative manufacturers of analytical measuring technology worldwide.

Our business unit Analytical Instrumentation offers excellent competencies in the fields of optical spectroscopy, sum parameters and elemental analysis. Being proud of our core competency we grant all our customers a long-term warranty of 10 years for our high-performance optics.

About Lab Automation

With more than 25 years of market experience, Analytik Jena with its CyBio® Product Line is a leading provider for high quality liquid handling and automation technologies. In the pharmaceutical and life science industries, our products enjoy the highest reputation for precision, reliability, robustness and simplicity.

Moreover, the Automation Team designs, produces and installs fully automated systems tailored to our clients' application, throughput and capacity requirements. From stand-alone CyBio® Well up to fully customized robotic systems we handle your compounds, biomolecules and cells with great care.


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Last updated: Feb 18, 2020 at 5:06 AM

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