iTRAQ proteomic analysis of aganglionic colonic mucosa to detect new diagnostic markers in Hirschsprung's disease

• Project/Area Number: 26461631
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Embryonic/Neonatal medicine
• Research Institution: Mie University
• Principal Investigator: INOUE Mikihiro 三重大学, 医学部附属病院, 助教 (30422835)
• Co-Investigator(Kenkyū-buntansha): 内田 恵一 三重大学, 医学部附属病院, 准教授 (30293781) ; 大竹 耕平 三重大学, 医学部, 助教 (40378344)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2016: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000); Fiscal Year 2015: ¥130,000 (Direct Cost: ¥100,000、Indirect Cost: ¥30,000); Fiscal Year 2014: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000)
• Keywords: ヒルシュスプルング病 / プロテオミクス / 診断マーカー

Outline of Final Research Achievements

Rectal mucosal biopsy is essential for the definite diagnosis of Hirschsprung’s disease (HD). However, this examination is sometimes less valuable when applied to neonates. Therefore, a less invasive and more accurate diagnostic approach is required for early screening and diagnosis of HD. In the present study, we used iTRAQ proteomic analysis to identify a novel candidate protein aimed to increase diagnostic accuracy. We prepared resected tissues from 4 patients with HD and 4 subject without HD (non-HD). The colonic mucosa of the aganglionic segment from the patient with HD, colonic mucosa of the ganglionic segment from the same HD patient and colonic mucosa from the non-HD subject were manually microdissected. The proteins were extracted, digested, labeled using iTRAQ reagents, and analyzed using mass spectrometer. We identified and quantitated 197 proteins. Among these proteins, the levels of 17 proteins were significantly up-regulated in the aganglionic HD sample.

Research Products

• [Presentation] iTRAQ proteomic analysis of formalin-fixed and paraffin-embedded aganglionic colonic mucosa of Hirschsprung disease: a pilot study (2016)
  • Author(s): 大竹耕平
  • Organizer: The 24th Congress of Asian Association of Pediatric Surgeons
  • Place of Presentation: ヒルトン福岡シーホーク（福岡県、福岡市）
  • Year and Date: 2016-05-24
  • Int'l Joint Research
• [Presentation] iTRAQ based proteomic analysis of formalin fixed and paraffin-embedded aganglionic colonic mucosa in Hirschsprung's disease: a pilot study (2016)
  • Author(s): Kohei Otake
  • Organizer: The 24th Congress of Asian Association of Pediatric Surgeons
  • Place of Presentation: ヒルトン福岡シーホーク（福岡県、福岡市）
  • Year and Date: 2016-05-22
  • Int'l Joint Research