RESEARCH PROPOSAL

Nathan Baker
Independent Research GT
2018

Title​: ​What is the Best way to Diagnose Transitional Cell Carcinoma as Early as Possible?

Introduction and Overview of Research:

Transitional Cell Carcinoma (TCC) is a highly aggressive cancer in dogs that is currently
almost always diagnosed in its later stages when it is hard to treat. The researcher discovered the
best way to screen TCC in dogs in its early stages is by using identification of TCC’s most
discriminating protein biomarkers. The researcher wants to use the findings from his original
study to create a general procedure for an ELISA. The researcher wants to get feedback on this
procedure from his advisor, Dr. Shay Bracha. The findings will be published for veterinarians
and veterinary oncologists to use. Ultimately, the procedure will act as a developmental base for
an ELISA that will be tested with an experiment in the future.
Background and Rationale
Transitional Cell Carcinoma (TCC) is a highly aggressive form of bladder cancer in dogs.
TCC forms malignant tumors, which can develop and spread very quickly. TCC is the most
common form of cancer that affects the urinary tract of dogs; it represents 1.2 - 2% of all cancers
in dogs (Glickman). Certain carcinogens have been shown to induce the disease. The increase of
carcinogens to which dogs are exposed to is causing TCC to become a growing problem. The
prevalence of TCC increased more than 600% as examined in North American teaching hospitals
from 1975 - 1995 (Glickman). This increase is still rapidly growing, making TCC even more of
an epidemic.
TCC is currently diagnosed with a biopsy and screened by the Bladder Tumor Antigen
test (BTA). The problem is symptoms of TCC appear in its later stages, when it is not the ideal
time to treat. Biopsies are only done when symptoms and a mass are present so biopsies cannot
identify TCC in its early stages. Another problem is Urinary Tract Infections (UTIs) have very
similar symptoms of TCC and the BTA test is not specific enough to effectively diagnose TCC
over a UTI. For these reasons, TCC needs a better diagnostic tool now.
The good news is that new knowledge is emerging on a way to use the proteins TCC
produces to indicate its existence. If a cheap way to identify the protein biomarkers of TCC was
discovered and was made accessible to animal clinics, TCC could be screened effectively in dogs
so it could be found in its early stages. An ELISA could be created that would use the antibodies
of the protein biomarkers of TCC and enzymes to produce a signal that would indicate the
existence of TCC. This could help present a inexpensive and effective way to diagnose TCC.

Research Question
What is the best way to screen Transitional Cell Carcinoma in dogs in its early stages?

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Research Thesis
Using protein identification of TCC’s unique, most discriminative protein biomarkers is
the best way to diagnose TCC in its early stages. Protein identification of TCC unique
biomarkers is 90% accurate (Bracha), nearly 100% specific, can be regularly screened, can be
tested at any stage, and with an ELISA it could be tested with minimal expense. These benefits
make it a great diagnostic tool, and if the protein identification could be done through a simple
ELISA it would be available for most animal clinics.
Research Design Model
Feedback will be generated from the researcher’s advisor, Dr. Shay Bracha. An ELISA is
an Enzyme Linked Immunosorbent Assay that produces a color change when a specific protein is
present. The procedure will be assessed based on multiple criteria including if it could be
reasonably made by veterinarians and if it has any flaws that would clearly result in false
positives or negatives.
Data Collection
In the original study, meta analysis was completed to find the accuracy and specificity of
diagnostic tools for TCC versus that of the investigational model of mass spectrometry of TCC’s
protein biomarkers. The researcher studied cystoscopic biopsies, BTA, mass spectrometry of
TCC unique biomarkers, and ​S100A8/A9USG. Data was collected from 4 previous experiments.
Meta analysis was the best choice to compare the accuracy and specificity of diagnostic tools
because the researcher could not run these experiments himself due to lack of resources. The
researcher discovered specific identification of TCC unique protein biomarkers is the best way to
diagnose TCC early due to the fact that it had one of the highest accuracy, very high specificity,
and the other benefits discovered in the literature review such as non-​invasiveness, ability to be
tested in any stage of TCC, ability to be regularly screened, and moderate expense​.
Increasing the availability of protein identification tools for TCC may change the way it
is screened in many clinics. The culmination of the data produced by the 4 studies provides a
direct comparison of the diagnostic tools studied for TCC. This has not been done before and it
provides a way for veterinarians and oncologists to weigh the pros and cons of each tool to
decide which is the best for their patients diagnosis. The literature review matched the data
collection, both found mass spectrometry of TCC unique protein biomarkers was overall the
most effective way to diagnose TCC early. It was also discovered that a benchmark assay could
reduce the cost of screening TCC’s unique protein biomarkers. All the finding influence the
product. The plan is to create a general procedure for an ELISA for TCC’s unique protein
biomarkers and provide the antibodies and enzyme for each protein if available. This can be
assessed by Dr. Shay Bracha and can be published for animal clinics or oncology labs.

Product Objectives:
The product will be a ​general procedure for an ELISA for TCC. ​This can be assessed by
Dr. Shay Bracha and can be published for animal clinics or oncology labs. ​The procedure will act
as a developmental base for an ELISA that will be tested with an experiment in the future.
If the ELISA is successful it will provide animal clinics or oncology labs the ability to create
their own ELISAs for minimal cost. The audience is veterinarians and veterinary oncologists
because they are the only people who can use the findings in a real world scenario. The products
and outcomes will be published online for the use of the audience of veterinarians and veterinary
oncologists.

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Logistical Considerations:
The only major logistical consideration for the procedure is feedback is necessary from
the researchers advisor, Dr Shay Bracha. When the product is later developed and tested there
will be additional logistical considerations.

Approval:

__________________________ ___________________________
_____________________
Student Signature G/T Resource Teacher Signature
Mentor/Advisor Signature

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 References

Bracha, S., McNamara, M., Hilgart, I., Milovancev, M., Medlock, J., Goodall, C., . . . Maier, C.
S. (2014, April). ​A Multiplex Biomarker Approach for the Diagnosis of Transitional Cell
Carcinoma from Canine Urine​. Retrieved from PubMed database.

Glickman, L. T., Raghavan, M., Knapp, D. W., Bonny, P. L., & Dawson, M. H. (2004, April 15).
Herbicide exposure and risk of transitional cell carcinoma of the urinary bladder in
scottish terriers​. Retrieved March 14, 2018, from
https://www.beyondpesticides.org/assets/media/documents/lawn/news/herbicides%20can
ine%20cancer.pdf