Intest Res.  2018 Jul;16(3):393-399. 10.5217/ir.2018.16.3.393.

Immunohistochemical differentiation between chronic enteropathy associated with SLCO2A1 gene and other inflammatory bowel diseases

• Yamaguchi
• Yanai S
• Nakamura S
• Kawasaki K
• Eizuka M
• Uesugi N
• Sugai T
• Umeno J
• Esaki M
• Matsumoto

• KMID: 2417651
• DOI: http://doi.org/10.5217/ir.2018.16.3.393

Abstract

BACKGROUND/AIMS
We recently identified recessive mutations in the solute carrier organic anion transporter family member 2A1 gene (SLCO2A1) as causative variants of chronic enteropathy associated with SLCO2A1 (CEAS). The aim of this study was to evaluate SLCO2A1 protein expression in the intestinal tissues of patients with CEAS, intestinal Behçet's disease (BD), simple ulcer (SU), and Crohn's disease (CD).
METHODS
Immunohistochemical staining using a polyclonal anti-SLCO2A1 antibody was performed on the resected intestinal specimens from 13 cases of CD, 9 cases of intestinal BD/SU, and 3 cases of CEAS. The extent of SLCO2A1 expression was determined by counting positively-staining vascular endothelial cells and scored as 0 (no cells), 1 (1%-30% cells), 2 (31%-60%), or 3 (>60%). The intensity of SLCO2A1 expression was scored either as 0 (negative), 1 (intermediate), or 2 (strong). The extent score and intensity score were summed for the final score of 0, 2, 3, 4, or 5.
RESULTS
SLCO2A1 protein expression was observed in 1 of 3 cases of CEAS (33%), all 13 cases of CD (100%), and all 9 cases of BD/SU (100%). The mean final expression scores of CEAS, CD, and BD/SU were 1.6 (range, 0-5), 4.8 (range, 4-5), and 4.3 (range, 4-5), respectively. The final expression score in CEAS was significantly lower than in CD (P=0.03).
CONCLUSIONS
Immunohistochemical staining of the SLCO2A1 protein is considered useful to distinguish CEAS from other inflammatory bowel diseases.

Keyword

SLCO2A1; Chronic enteropathy; Crohn disease; Behcet syndrome; Immunohistochemistry

MeSH Terms

Behcet Syndrome
Crohn Disease
Endothelial Cells
Humans
Immunohistochemistry
Inflammatory Bowel Diseases*
Ulcer

Figure

• 

  Fig. 1 Examples of SLCO2A1 expression intensity scores of vascular endothelial cells. (A) Strong (score 2). (B) Intermediate (score 1). (C) Negative (score 0). Dako Envision detection kit, original magnification, ×200.

• 

  Fig. 2 (A) Immunostaining of SLCO2A1 in a case of CD (57 years old at surgery, male). Submucosal areas in normal-appearing ileal tissue adjacent to the ulcerative lesion. Extent score=3, intensity score=2, final score=5 (original magnification, ×100). (B, C) Immunostaining of SLCO2A1 in a case of BD (60 years old at surgery, female, original magnification, ×400). (B) Tissues from ulcerative lesion in the submucosa. Extent score=3, intensity score=1, final score=4. (C) Submucosal areas of normal-appearing colonic tissue adjacent to the ulcerative lesions. Extent score=3, intensity score=2, final score=5. (D-F) Immunostaining of SLCO2A1 in 3 cases of CEAS (original magnification, ×200). (D) Case 1: 34 years old at diagnosis, 50 years old at surgery, female. Extent score=0, intensity score=0, final score=0. (E) Case 2: 22 years old at diagnosis, 40 years old at surgery, female. Extent score=0, intensity score=0, final score=0. (F) Case 3: 22 years old at diagnosis, 27 years old at surgery, female. Extent score=3, intensity score=2, final score=5. CEAS, chronic enteropathy associated with SLCO2A1.

• 

  Fig. 3 The scores of SLCO2A1 expression in each group. (A) Extent scores: significant difference was observed between cases of CD and CEAS (P<0.05). (B) Intensity scores: significant difference was observed between cases of CD and CEAS (P<0.005), and between cases of BD/S and CEAS (P<0.05). (C) Final scores: significant difference was observed between cases of CD and CEAS (P<0.05), but not between cases of BD/SU and CEAS (P=0.23). BD, Behçet's disease; SU, simple ulcer; CEAS, chronic enteropathy associated with SLCO2A1.

• 

  Fig. 4 Final scores of SLCO2A1 staining in normal-appearing tissues and ulcerative lesions in cases of BD/SU. A significant difference was observed between the 2 groups (P<0.05). BD, Behçet's disease; SU, simple ulcer.

• 

  Fig. 5 Structures of wild-type SLCO2A1 proteins and CEAS mutations. While the full length of the SLCO2A1 protein is 643 amino acids (AAs), the antibody HPA013742 recognizes the 83 AAs of the 5th extracellular domain coded by exons 9-11 (431-513, http://www.proteinatlas.org/ENSG00000174640-SLCO2A1/antibody). In cases 1 and 2, the length of the synthesized SLCO2A1 protein was considered too short for detection by this antibody. Case 3 had the compound heterozygous mutation and the full length SLCO2A1 protein was synthesized and detected by HPA013742.

Reference

1. Matsumoto T, Iida M, Matsui T, et al. Non-specific multiple ulcers of the small intestine unrelated to non-steroidal anti-inflammatory drugs. J Clin Pathol. 2004; 57:1145–1150. PMID: 15509673.

2. Matsumoto T, Iida M, Matsui T, Yao T. Chronic nonspecific multiple ulcers of the small intestine: a proposal of the entity from Japanese gastroenterologists to Western enteroscopists. Gastrointest Endosc. 2007; 66(3 Suppl):S99–S107. PMID: 17709045.

3. Matsumoto T, Kubokura N, Matsui T, Iida M, Yao T. Chronic nonspecific multiple ulcer of the small intestine segregates in offspring from consanguinity. J Crohns Colitis. 2011; 5:559–565. PMID: 22115375.

4. Esaki M, Umeno J, Kitazono T, Matsumoto T. Clinicopathologic features of chronic nonspecific multiple ulcers of the small intestine. Clin J Gastroenterol. 2015; 8:57–62. PMID: 25788296.

5. Umeno J, Hisamatsu T, Esaki M, et al. A hereditary enteropathy caused by mutations in the SLCO2A1 gene, encoding a prostaglandin transporter. PLoS Genet. 2015; 11:e1005581. DOI: 10.1371/journal.pgen.1005581. PMID: 26539716.

6. Zhang Z, Xia W, He J, et al. Exome sequencing identifies SLCO2A1 mutations as a cause of primary hypertrophic osteoarthropathy. Am J Hum Genet. 2012; 90:125–132. PMID: 22197487.

7. Busch J, Frank V, Bachmann N, et al. Mutations in the prostaglandin transporter SLCO2A1 cause primary hypertrophic osteoarthropathy with digital clubbing. J Invest Dermatol. 2012; 132:2473–2476. PMID: 22696055.

8. Diggle CP, Parry DA, Logan CV, et al. Prostaglandin transporter mutations cause pachydermoperiostosis with myelofibrosis. Hum Mutat. 2012; 33:1175–1181. PMID: 22553128.

9. Compton RF, Sandborn WJ, Yang H, et al. A new syndrome of Crohn's disease and pachydermoperiostosis in a family. Gastroenterology. 1997; 112:241–249. PMID: 8978365.

10. Hisabe T, Hirai F, Matsui T, Watanabe M. Evaluation of diagnostic criteria for Crohn's disease in Japan. J Gastroenterol. 2014; 49:93–99. PMID: 23546557.

11. Hisamatsu T, Ueno F, Matsumoto T, et al. The 2nd edition of consensus statements for the diagnosis and management of intestinal Behçet's disease: indication of anti-TNFalpha monoclonal antibodies. J Gastroenterol. 2014; 49:156–162. PMID: 23955155.

12. Muto T. Historical review of so-called “simple ulcer” of the intestine. Stomach Intestine. 1979; 14:739–749.

13. Matsukawa M, Yamasaki T, Kouda T, Kurihara M. Endoscopic therapy with absolute ethanol for postoperative recurrent ulcers in intestinal Behçe's disease, and simple ulcers. J Gastroenterol. 2001; 36:255–258. PMID: 11324729.

14. Allred DC, Clark GM, Elledge R, et al. Association of p53 protein expression with tumor cell proliferation rate and clinical outcome in node-negative breast cancer. J Natl Cancer Inst. 1993; 85:200–206. PMID: 8423624.

15. Harvey JM, Clark GM, Osborne CK, Allred DC. Estrogen receptor status by immunohistochemistry is superior to the ligand-binding assay for predicting response to adjuvant endocrine therapy in breast cancer. J Clin Oncol. 1999; 17:1474–1481. PMID: 10334533.

16. Shim YW, Suh JS. Primary hypertrophic osteoarthropathy accompanied by Crohn's disease: a case report. Yonsei Med J. 1997; 38:319–322. PMID: 9409195.

17. McAllister C, McNulty JG, Fielding JF. Is hypertrophic osteoarthropathy really so rare in regional enteritis? J Clin Gastroenterol. 1986; 8:562–566. PMID: 3782755.