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. 2021 Jan;35(1):179-189.
doi: 10.1111/jvim.16003. Epub 2021 Jan 20.

Characterization of the intestinal mucosal proteome in cats with inflammatory bowel disease and alimentary small cell lymphoma

Sina Marsilio  1   2 , Floris C Dröes  2 , Lawrence Dangott  3 , Betty Chow  4   5 , Steve Hill  4   6 , Mark Ackermann  7 , J Scott Estep  8 , Jonathan A Lidbury  2 , Jan S Suchodolski  2 , Jörg M Steiner  2
Affiliations

Characterization of the intestinal mucosal proteome in cats with inflammatory bowel disease and alimentary small cell lymphoma

Sina Marsilio et al. J Vet Intern Med. 2021 Jan.

Erratum in

Abstract

Background: Current tests for diagnosis and differentiation of lymphoplasmacytic enteritis (LPE) and small cell lymphoma (SCL) in cats are expensive, invasive, and lack specificity. The identification of less invasive, more reliable biomarkers would facilitate diagnosis.

Objectives: To characterize the mucosal proteome in endoscopically obtained, small intestinal tissue biopsy specimens. We hypothesized that differentially expressed proteins could be identified and serve as biomarker candidates for the differentiation of LPE and SCL in cats.

Animals: Six healthy control cats, 6 cats with LPE, and 8 cats with SCL.

Methods: The mucosal proteome was analyzed using 2-dimensional fluorescence difference gel electrophoresis (2D DIGE) and nanoflow liquid chromatography tandem mass spectrometry. For 5 proteins, results were verified by Western blot analysis.

Results: A total of 2349 spots were identified, of which 9 were differentially expressed with a ≥2-fold change between healthy cats and cats with LPE and SCL (.01 < P < .001). Eight of these 9 spots were also differentially expressed between cats with LPE and cats with SCL (P .001 < P < .04). However, Western blot analysis for malate dehydrogenase-1, malate dehydrogenase-2, apolipoprotein, annexin IV, and annexin V did not confirm significant differential protein expression for any of the 5 proteins assessed.

Conclusions and clinical importance: Two-D DIGE did not identify potential biomarker candidates in the intestinal mucosa of cats with LPE and SCL. Future studies should focus on different techniques to identify biomarker candidates for cats with chronic enteropathies (CE).

Keywords: EATL; enteropathy-associated T-cell lymphoma; feline chronic enteropathy.

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Conflict of interest statement

At the time of the study, authors Marsilio, Dröes, Lidbury, Suchodolski, and Steiner are or were employed by the Gastrointestinal Laboratory at Texas A&M University, which offers laboratory testing, including histopathology services, on a fee‐for‐service basis. The author Dangott is an employee of the Protein Chemistry Laboratory at Texas A&M, which offers laboratory testing, including 2D DIGE, on a fee‐for‐service basis. The author Ackermann is affiliated with the Gastrointestinal Laboratory at Texas A&M University. The author Estep is employed by Texas Veterinary Pathology, LLC, which offers histopathology for animals on a fee‐for‐service basis. The authors Chow and Hill have nothing to disclose.

Figures

FIGURE 1
FIGURE 1
Representative examples of a 2‐dimensional difference in gel electrophoresis (2D‐DIGE) imaged using the ImageQuant software and analyzed by the DeCyder software. A, Representative fluorescent protein profiles of a gel containing protein samples extracted from the small intestinal mucosa of a cat with lymphoplasmacytic enteritis (LPE) labeled with Cy2 (top left), a cat with intestinal small cell lymphoma (SCL) labeled with Cy5 (top middle), a pooled internal control labeled with Cy3 (top right), and the overlay image (bottom) as seen in the ImageQuant software. B. Representative view of gel images uploaded into DeCyder software and analyzed by the Biological Variation Analysis module. Top: black and white images of gels containing protein samples from a cat with LPE (left) and SCL (right). Bottom: Enlarged region and 3‐D view of upregulated spot 1065 within the respective gels. The pink area demarcates the area analyzed by DeCyder for protein spot intensity. The yellow cylinder represents the area that can be picked during spot picking and analyzed by mass spectrometry for protein identification
See this image and copyright information in PMC

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