Fragmentations of transthyretin in familial amyloid polyneuropathy

• Project/Area Number: 25870541
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Laboratory medicine; Neurology
• Research Institution: Kumamoto University
• Principal Investigator: UEDA Mitsuharu 熊本大学, 医学部附属病院, 講師 (60452885)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2013: ¥2,990,000 (Direct Cost: ¥2,300,000、Indirect Cost: ¥690,000)
• Keywords: トランスサイレチン / アミロイドーシス / 難病 / FAP / 断片化 / トリプシン

Outline of Final Research Achievements

Fragmentations of transthyretin (TTR) were reportedly found in amyloid deposits in familial amyloid neuropathy (FAP). However, the mechanism and pathological roles of the TTR fragmentations in FAP remain to be determined. To elucidate the mechanism involved in TTR fragmentations, we investigated how TTR fragments were formed in cultured cells.
Recombinat TTR was employed in this study. Several kinds of cultured cells were incubated for 24 hours with soluble TTR or TTR aggregates formed in vitro by pretreating in an acidic buffer. After incubation, cell lysates were examined by SDS-PAGE and amino-acid sequence analysis to detect TTR fragments. TTR fragments were detected in lysates of neuron (SH-SY5H) and astrocyte (U87MG) cell lines which were incubated with TTR aggregates, but not in those which were incubated with soluble TTR. Amino-acid sequence analysis revealed that cleavages of TTR were made at C-terminal portions.

Research Products

• [Journal Article] Clinico-histopathological and biochemical analyses of corneal amyloidosis in gelatinous drop-like corneal dystrophy. (2015)
  • Author(s): Tasaki M, Ueda M, Matsumoto K, Kawaji T, Misumi Y, Eiki D, Suenaga G, Obayashi K, Yamashita T, Tanihara H, Ando Y.
  • Journal Title: Amyloid Volume: 22 Issue: 1 Pages: 67-69
  • DOI: 10.3109/13506129.2014.983222
  • Peer Reviewed
• [Journal Article] Amyloid deposits derived from transthyretin in the ligamentum flavum as related to lumbar spinal canal stenosis. (2015)
  • Author(s): Yanagisawa A, Ueda M, Sueyoshi T, Okada T, Fujimoto T, Ogi Y, Kitagawa K, Tasaki M, Misumi Y, Oshima T, Jono H, Obayashi K, Hirakawa K, Uchida H, Westermark P, Ando Y, Mizuta H.
  • Journal Title: Mod Pathol Volume: 28 Issue: 2 Pages: 201-207
  • DOI: 10.1038/modpathol.2014.102
  • Peer Reviewed
• [Journal Article] Osteopontin-integrin α(v)β(3) axis is crucial for 5-fluorouracil resistance in oral squamous cell carcinoma. (2015)
  • Author(s): Nakamura T, Shinriki S, Jono H, Ueda M, Nagata M, Guo J, Hayashi M, Yoshida R, Ota T, Ota K, Kawahara K, Nakagawa Y, Yamashita S, Nakayama H, Hiraki A, Shinohara M, Ando Y.
  • Journal Title: FEBS Lett Volume: 589 Issue: 2 Pages: 231-239
  • DOI: 10.1016/j.febslet.2014.12.004
  • Peer Reviewed / Open Access
• [Journal Article] Intrinsic TGF-β2-triggered SDF-1-CXCR4 signaling axis is crucial for drug resistance and a slow-cycling state in bone marrow-disseminated tumor cells. (2015)
  • Author(s): Nakamura T, Shinriki S, Jono H, Guo J, Ueda M, Hayashi M, Yamashita S, Zijlstra A, Nakayama H, Hiraki A, Shinohara M, Ando Y.
  • Journal Title: Oncotarget Volume: 6 Pages: 1008-1019
  • Peer Reviewed
• [Journal Article] Shikonin, dually functions as a proteasome inhibitor and a necroptosis inducer in multiple myeloma cells. (2015)
  • Author(s): Wada N, Kawano Y, Fujiwara S, Kikukawa Y, Okuno Y, Tasaki M, Ueda M, Ando Y, Yoshinaga K, Ri M, Iida S, Nakashima T, Shiotsu Y, Mitsuya H, Hata H.
  • Journal Title: International Journal of Oncology Volume: 46 Issue: 3 Pages: 963-972
  • DOI: 10.3892/ijo.2014.2804
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Megalencephalic leukoencephalopathy with subcortical cysts caused by compound heterozygous mutations in MLC1, in patients with and without subcortical cysts in the brain. (2015)
  • Author(s): Masuda T, Ueda M, Ueyama H, Shimada S, Ishizaki M, Imamura S, Yamamoto T, Ando Y.
  • Journal Title: J Neurol Sci Volume: 351 Issue: 1-2 Pages: 211-213
  • DOI: 10.1016/j.jns.2015.03.010
  • Peer Reviewed
• [Journal Article] Contactin 1 IgG4 associates to chronic inflammatory demyelinating polyneuropathy with sensory ataxia. (2015)
  • Author(s): Miura Y, Devaux JJ, Fukami Y, Manso C, Belghazi M, Wong AH, Yuki N, CNTN1-CIDP Study Group (Fujimura H, Toshio Fukutake T, Iwanami H, Kusaka H, Kuwabara S, Okuma Y, Ueda M, Yamamoto T)
  • Journal Title: Brain Volume: Epub
  • Peer Reviewed
• [Journal Article] Myopathic phenotype of familial amyloid polyneuropathy with a rare transthyretin variant: ATTR Ala45Asp. (2014)
  • Author(s): Misumi Y, Doki T, Ueda M, Obayashi K, Tasaki M, Tamura A, Ando Y.
  • Journal Title: Amyloid Volume: 21 Issue: 3 Pages: 216-217
  • DOI: 10.3109/13506129.2014.932277
  • Peer Reviewed
• [Journal Article] Hypoxia suppresses cylindromatosis (CYLD) expression to promote inflammation in glioblastoma: possible link to acquired resistance to anti-VEGF therapy. (2014)
  • Author(s): Guo J, Shinriki S, Su Y, Nakamura T, Hayashi M, Tsuda Y, Murakami Y, Tasaki M, Hide T, Takezaki T, Kuratsu J, Yamashita S, Ueda M, Li J, Ando Y, Jono H.
  • Journal Title: Oncotarget Volume: 5 Pages: 6353-6364
  • Peer Reviewed / Open Access
• [Journal Article] Juvenile-onset parkinsonism with digenic parkin and PINK1 mutations treated with subthalamic nucleus stimulation at 45 years after disease onset. (2014)
  • Author(s): Nakahara K, Ueda M, Yamada K, Koide T, Yoshimochi G, Funayama M, Kim JH, Yamakawa S, Mori A, Misumi Y, Uyama E, Hattori N, Ando Y.
  • Journal Title: J Neurol Sci. Volume: 15 Issue: 1-2 Pages: 276-277
  • DOI: 10.1016/j.jns.2014.07.053
  • Peer Reviewed
• [Journal Article] Generation of familial amyloidotic polyneuropathy-specific induced pluripotent stem cells. (2014)
  • Author(s): Isono K, Jono H, Ohya Y, Shiraki N, Yamazoe T, Sugasaki A, Era T, Fusaki N, Tasaki M, Ueda M, Shinriki S, Inomata Y, Kume S, Ando Y.
  • Journal Title: Stem Cell Res Volume: 12 Issue: 2 Pages: 574-83
  • DOI: 10.1016/j.scr.2014.01.004
  • Peer Reviewed
• [Journal Article] Stromal Expression of Neutrophil Gelatinase-Associated Lipocalin Correlates with Poor Differentiation and Adverse Prognosis in Oral Squamous Cell Carcinoma. (2014)
  • Author(s): Shinriki S., Jono H., Ueda M., Obayashi K., Nakamura T., Ota K., Ota T., Sueyoshi T., Guo J., Hayashi M., Hiraki A., Nakayama H., Yamashita S., Shinohara M., & Ando Y.
  • Journal Title: Histopathology Volume: 64 Issue: 3 Pages: 353-364
  • DOI: 10.1111/his.12293
  • Peer Reviewed
• [Journal Article] Changes in pathological and biochemical findings of systemic tissue sites in familial amyloid polyneuropathy more than 10 years after liver transplantation (2013)
  • Author(s): Oshima T, Kawahara S, Ueda M, Kawakami Y, Tanaka R, Okazaki T, Misumi Y, Obayashi K, Yamashita T, Ohya Y, Ihse E, Shinriki S, Tasaki M, Jono H, Asonuma K, Inomata Y, Westermark P, Ando Y
  • Journal Title: J Neurol Neurosurg Psychiatry Volume: - Issue: 7 Pages: 740-746
  • DOI: 10.1136/jnnp-2013-305973
  • Peer Reviewed
• [Journal Article] Effect of age and sex differences on wild-type transthyretin amyloid formation in familial amyloidotic polyneuropathy: a proteomic approach (2013)
  • Author(s): Tasaki M, Ueda M, Obayashi K, Koike H, Kitagawa K, Ogi Y, Jono H, Su Y, Suenaga G, Oshima T, Misumi Y, Yoshida M, Yamashita T, Sobue G, Ando Y
  • Journal Title: Int J Cardiol Volume: 170 Issue: 1 Pages: 69-74
  • DOI: 10.1016/j.ijcard.2013.10.033
  • Peer Reviewed
• [Journal Article] Dominant cardiac type of familial amyloidotic polyneuropathy associated with a novel transthyretin variant Thr59Arg (2013)
  • Author(s): Obayashi K, Misumi Y, Watanabe T, Ando T, Akagami T, Tasaki M, Shinriki S, Ueda M, Yamashita T, Hirotani S, Ando Y
  • Journal Title: Muscle Nerve Volume: 48(5) Issue: 5 Pages: 839-41
  • DOI: 10.1002/mus.23939
  • Peer Reviewed
• [Journal Article] Marked cardiomegaly in a patient with familial amyloidotic polyneuropathy after orthotopic liver transplantation: a case study (2013)
  • Author(s): Sakashita N, Kagawa S, Date R, Ueno T, Nakagawa T, Yamashita T, Obayashi K, Ohshima T, Ueda M, Horiguchi H, Ando Y, Takeya M
  • Journal Title: Pathol Int Volume: 63 Issue: 5 Pages: 260-265
  • DOI: 10.1111/pin.12055
  • Peer Reviewed
• [Journal Article] Amyloid fibrils containing fragmented ATTR may be the standard fibril composition in ATTR amyloidosis (2013)
  • Author(s): Ihse E, Rapezzi C, Merlini G, Benson MD, Ando Y, Suhr OB, Ikeda S, Lavatelli F, Obici L, Quarta CC, Leone O, Jono H, Ueda M, Lorenzini M, Liepnieks J, Ohshima T, Tasaki M, Yamashita T, Westermark P
  • Journal Title: Amyloid Volume: 20(3) Issue: 3 Pages: 142-50
  • DOI: 10.3109/13506129.2013.797890
  • Peer Reviewed
• [Journal Article] Impact of antibodies against amyloidogenic transthyretin (ATTR) on phenotypes of patients with familial amyloidotic polyneuropathy (FAP) ATTR Valine30Methionine (2013)
  • Author(s): Obayashi K, Tasaki M, Jono H, Ueda M, Shinriki S, Misumi Y, Yamashita T, Oshima T, Nakamura T, Ikemizu S, Anan I, Suhr O, Ando Y
  • Journal Title: Clin Chim Acta Volume: 419 Pages: 127-31
  • DOI: 10.1016/j.cca.2013.02.002
  • Peer Reviewed
• [Presentation] Amyloidogenicity of transthyretin and diseases. (2015)
  • Author(s): Ueda M, Ando Y.
  • Organizer: The 10th Japan-China Cooperative Life Science Symposium.
  • Place of Presentation: Kumamoto, Japan
  • Year and Date: 2015-10-27
  • Invited
• [Presentation] 質量分析法によるアミロイドーシス診断法の確立． (2014)
  • Author(s): 植田光晴、田崎雅義、荻泰裕、北川敬資、柳澤哲大、井上泰輝、三隅洋平、増田曜章、黄冠男、久原春代、大隈雅紀、池田勝義、大林光念、山下太郎、安東由喜雄
  • Organizer: 第61回日本臨床検査医学会学術集会
  • Place of Presentation: 福岡
  • Year and Date: 2014-11-22 – 2014-11-25
• [Presentation] トランスサイレチン断片化機構およびアミロイド形成へ与える影響の解析． (2014)
  • Author(s): 植田光晴、豊島梨沙、水口峰之、Barbara Kluve-Beckerman、Juris J. Liepnieks、田崎雅義、三隅洋平、山下太郎、Merrill D. Benson、安東由喜雄
  • Organizer: 第87回日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 2014-10-15 – 2014-10-18
• [Presentation] トランスサイレチンの断片化機構の解析と病態への関与． (2014)
  • Author(s): 植田光晴、水口 峰之、安東 由喜雄
  • Organizer: 第2回アミロイドーシス研究会学術集会
  • Place of Presentation: 東京
  • Year and Date: 2014-08-22
  • Invited
• [Presentation] 肝移植後に長期経過した家族性アミロイドポリニューロパチー患者の病理学的検討． (2014)
  • Author(s): 植田光晴、大嶋俊範、三隅洋平、山下太郎、大林光念、安東由喜雄
  • Organizer: 第55回日本神経病理学会総会学術研究会
  • Place of Presentation: 東京
  • Year and Date: 2014-06-05 – 2014-06-07
• [Presentation] FAP発症年齢に影響するTTR断片化生成機構の解析． (2014)
  • Author(s): 植田光晴、Barbara Kluve-Beckerman、Juris J. Liepnieks、大嶋俊範、水口峰之、三隅洋平、大林光念、Merrill D. Benson、安東由喜雄
  • Organizer: 第55回日本神経学会学術大会
  • Place of Presentation: 福岡
  • Year and Date: 2014-05-21 – 2014-05-24
• [Presentation] Fragmentations of TTR in cultured cells. (2014)
  • Author(s): Ueda M, Kluve-Beckerman B, Liepnieks J, Mizuguchi M, Misumi Y, Ando Y, Benson M.
  • Organizer: The XIVth International Symposium on Amyloidosis.
  • Place of Presentation: Indianapolis, IN, USA
  • Year and Date: 2014-04-27 – 2014-05-01
• [Presentation] Fragmentation of amyloid-forming transthyretin in cultured cells and a test tube. (2013)
  • Author(s): Ueda M, Kluve-Beckerman B, Liepnieks J, Mizuguchi M, Ando Y, Benson M.
  • Organizer: The VIIIth International Symposium on Familial Amyloidotic Polyneuropathy.
  • Place of Presentation: Rio de Janeiro, Brazil
• [Presentation] FAPの病態の鍵を握るトランスサイレチン断片化機構の解析． (2013)
  • Author(s): 植田光晴、Barbara Kluve-Beckerman、Juris J. Liepnieks、水口峰之、安東由喜雄、Merrill D. Benson．
  • Organizer: 第54回日本神経学会総会
  • Place of Presentation: 東京
• [Presentation] FAP病態の鍵を握るトランスサイレチン断片化機構の解明と検出 (2013)
  • Author(s): 植田 光晴、Barbara Kluve-Beckerman、Juris J. Liepnieks、田崎 雅義、大林 光念、Merrill D. Benson、安東 由喜雄
  • Organizer: 第60回 日本臨床検査医学会学術集会
  • Place of Presentation: 神戸
• [Book] アミロイドーシス検査における質量分析 (2013)
  • Author(s): 植田光晴、安東由喜雄
  • Publisher: 診断と治療社