TRYPANASOMAGRUZI : MOLECULAR BASES FOR INTRACELLULAR PARASITISM, PROLIFERATION, AND PATHOGENICITY IN INFECTED HUMAN CELLS

2007 Fiscal Year Final Research Report Summary

• Project/Area Number: 17390123
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Parasitology (including Sanitary zoology)
• Research Institution: Juntendo University
• Principal Investigator: AOKI Takashi Juntendo University, SCH. MED, PROFESSOR (20053283)
• Co-Investigator(Kenkyū-buntansha): NARA TAKESHI JUNTENDO UNIVERSITY, SCH. MED, ASSIST PROFESSOR (40276473) ; HASHIMOTO Muneaki JUNTENDO UNIVERSITY, SCH. MED, ASSOC. PROFESSOR (30407308) ; ANNOURA TAKESHI JUNTENDO UNIVERSITY, SCR MED, ASSIST PROFESSOR (90407239) ; TSUBOUCHI AKIKO JUNTENDO UNIVERSITY, SCH. MED, ASSIST PROFESSOR (10398662)
• Project Period (FY): 2005 – 2007
• Keywords: TRYPANOSOMA CRUZI / INFECTED HUMAN CELLS / INHIBITION OF APOPTOSIS / UBIQUITINATION OF cFLIP / CHAGAS' DISEASE / PYRIMIDINE -BIOSYNTHETIC GENE CLUSTER / TRANSCRIPTOME / UPREGULATION OF PROLIFERATION INHIBITORS

Research Abstract

Intracellular persistence of the protozoan parasite, Trypanosoma cruzi, is an aggravating cause of Chagas' disease, involving that the protozoan infection specifically inhibits death receptor-mediated apoptosis of host cells. We previously demonstrated that the parasite dramatically upregulates cellular FLICE inhibitory protein (cFLIP) , the only known mammalian inhibitor specific for death receptor signaling, in infected cells. While T cruzi infected cells had an expression level of Itch which is suggested to be an ubiquitin ligase for cFLIPL equivalent to that in uninfected cells, co-immunoprecipitation analysis revealed that the interaction between cFLIPL and Itch is strongly inhibited in T cruzi infected cells. This unique parasite strategy, which has not been reported in any pathogen-infected cells, may allow the host cell to accumulate cFLIPL, eventually resulting in the inhibition of apoptosis of T cruzi infected cells.
Upon an exhaustive transcriptome analysis of T cruzi infected HeLa cells in comparison with uninfected cells, the former showed >3-fold up-regulation of 41 genes and >3-fold down-regulation of 23 genes. Among these genes, seven up-regulated genes encode proliferation inhibitors and three down-regulated genes encode proliferation promoters, strongly. Suggesting that T cruzi infection inhibits host cell proliferation, which may allow more time for T cruzi to replicate and produce its intracellular nests.
In the diplonemid Diplonema papillatum that belongs to the sister group of kinetoplastids, phylogenetic analyses of pyr4 and pyr6 genes showed the separate origin of each in kinetoplastids and euglenoids/diplonemids and suggested that kinetoplastids have acquired these genes via lateral gene transfer. Taken together, we expect that the progress in the present study on "Trypanosoma cruzi: Molecular bases for intracellular parasitism, proliferation, and pathogenicity" would be extended to an applied study in near future.

Research Products

• [Journal Article] Transcriptome profile of Trypanosoma cruzi-infected cells:simultaneous up-and down-regulation of proliferation inhibitors and promoters (2008)
  • Author(s): Shigihara T, Hashimoto M, Shindo N, Aoki T
  • Journal Title: Parasitology Research (印刷中)
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Evolutionary analysis of synteny and gene fusion for pyrimidine biosynthetic enzymes in Euglenozoa:the gap between kinetoplastids and diplonemids (2008)
  • Author(s): Makiuchi T, Annoura T, Hashimoto T, Murata E, Aoki T, Nara T
  • Journal Title: Protist (印刷中)
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Transcriptome profile of Trypanosoma cruzrinfected cells : simultaneous up- and down-regulation of proliferation inhibitors and promoters (2008)
  • Author(s): Shigihara, T., Hashimoto, M., Shindo, N., Aoki, T
  • Journal Title: Parasitol Res (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Evolutionary analysis of synteny and gene fusion for pyrimidine biosynthetic enzymes in Euglenozoa : an extraordinary gap between kinetoplastids and diplonemids (2008)
  • Author(s): Makiuchi, T., Annoura, T., Hashimoto, T., Murata, E., Aoki, T., Nara, T
  • Journal Title: Protist (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Occurrence of multiple,independent gene fusion events for the fifth and sixth enzymes of pyrimidine biosynthesis in different eukaryotic groups (2007)
  • Author(s): Makiuchi T, Nara T, Annoura T, Hashimoto T, Aoki T
  • Journal Title: Gene 394 Pages: 78-86
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Dihydroorotate dehydrogenase arises from novel fused gene product with aspartate carbamoyltransferase in Bodo saliens (2007)
  • Author(s): Annoura T, Sariego I, Nara T, Makiuchi T, Fujimura T, Taka H, Mineki R, Murayama K, Aoki T
  • Journal Title: Biochemical and Biophysical Research Communications 358 Pages: 253-258
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Occurrence of multiple, independent gene fusion events for the fifth and sixth enzymes of pyrimidine biosynthesis in different eukaryotic groups (2007)
  • Author(s): Makiuchi, T., Nara, T., Annoura, T., Hashimoto, T., Aoki, T
  • Journal Title: Gene 394 Pages: 78-86
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Dihydroorotate dehydrogenase arises from novel fused gene product with aspartate carbamoyl-transferase in Bodo saliens (2007)
  • Author(s): Annoura, T., Sariego, I., Nara, T., Makiuchi, T., Fujimura, T., Taka, H., Mineki, R, Murayama, K., Aoki, T
  • Journal Title: Biochem Biophys Res Comm un 358 Pages: 253-258
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Genetic diversity and kinetic properties of Trypanosoma cruzi dihydroorotate dehydrogenase isoforms (2006)
  • Author(s): Sariego I, Annoura T, Nara T, Hashimoto M, Tsubouchi A, Iizumi K, Makiuchi T, Murata E, Kita K, Aoki T
  • Journal Title: Parasitology International 55 Pages: 11-16
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Molecular cloning and characterization of ouabain-insensitive Na^+-ATPase in the parasitic protist,Trypanosoma cruzi (2006)
  • Author(s): Iizumi K, Mikami Y, Hashimoto M, Nara T, Hara Y, Aoki T
  • Journal Title: Biochimica et Biophysica Acta-Biomembranres 1758 Pages: 738-746
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Genetic diversity and kinetic properties of Trypanosoma cruzi dihydroorotate dehydrogenase isoforms (2006)
  • Author(s): Sariego, I., Annoura, T., Nara, T., Hashimoto, M., Tsubouchi, A., Iizumi, K., Makiuchi, T., Murata, E., Kita, K., Aoki, T
  • Journal Title: Parasitollnt 55 Pages: 11-16
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Molecular cloning and characterization of ouabain-insensitive Na+-ATPase in the parasitic protist, Trypanosoma cruzi (2006)
  • Author(s): Iizumi, K., Mikami, Y., Hashimoto, M., Nara, T., Hara, Y., Aoki
  • Journal Title: Biochim BiophysActa Biomembranes 1758 Pages: 738-746
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Trypanosoma cruzi posttranscriptionally up-regulates and exploits cellular FLIP for inhibition of death-inducing signal (2005)
  • Author(s): Hashimoto M, N.-Shimada J, Aoki T
  • Journal Title: Molecular Biology of the Cell 16 Pages: 3521-3528
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Gene expression profiles in response to Fas stimulation in Trypanosoma cruzi-infected host cells (2005)
  • Author(s): Hashimoto M, Nakajima-Shimada J, Ishidoh K, Aoki T
  • Journal Title: International Journal for Parasitology 35 Pages: 1587-1594
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] The origin of dihydroorotate dehydrogenase genes of kinetoplastids,with special reference to their biological significance and adaptation to anaerobic,parasitic conditions (2005)
  • Author(s): Annoura T, Nara T, Makiuchi T, Hashimoto T, Aoki T
  • Journal Title: Journal of Molecular Evolution 60 Pages: 113-127
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Inhibitory action of marine algae extracts on the Trypanosoma cruzi dihydroorotate dehydrogenase activity and on the protozoan growth in mammalian cells (2005)
  • Author(s): Nara T, Kamei Y, Tsubouchi A, Annoura T, Hirota K, Iizumi K, Dohmoto Y, Ono T, Aoki T
  • Journal Title: Parasitology International 54 Pages: 59-64
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Expression,purification and crystallization of Trypanosoma cruzi dihydroorotate dehydrogenase complexed with orotate (2005)
  • Author(s): Inaoka DK, Takashima E, Osanai A, Shimizu H, Nara T, Aoki T, Harada S, Kita K
  • Journal Title: Acta Crystallographica F61 Pages: 875-878
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Antichagasic activity of komaroviquinone is due to generation of reactive oxygen species catalyzed by Trypanosoma cruzi old yellow enzyme (2005)
  • Author(s): Uchiyama N, Kabututu Z, Kubata BK, Kiuchi F, Ito M, Nakajima-Shimada J, Aoki T, Ohkubo K, Fukuzumi S, Martin SK, Honda G, Urade Y
  • Journal Title: Antimicrobial Agents and Chemotherapy 49 Pages: 5123-5126
  • Description: 「研究成果報告書概要(和文)」より
  • Peer Reviewed
• [Journal Article] Trypanosoma cruzi post-transcriptionally upregulates and exploits cellular FLIP for inhibition of death-inducing signal (2005)
  • Author(s): Hashimoto, M., N.-Shimada, J., Aoki, T
  • Journal Title: Mol Biol Cell 16 Pages: 3521-3528
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Gene Expression profiles in response to Fas stimulation in Trypanosoma cruzrinfected host cells (2005)
  • Author(s): Hashimoto, M., Nakajima-Shimada, J., Ishidoh, K., Aoki, T
  • Journal Title: Int J Parasitol 35 Pages: 1587-1594
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The origin of dihydroorotate dehydrogenase genes of kinetoplastids, with special reference to their biological significance and adaptation to anaerobic, parasitic conditions (2005)
  • Author(s): Annoura, T., Nara, T., Makiuchi, T., Hashimoto, T., Aoki, T
  • Journal Title: JMol Evol 60 Pages: 113-127
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Inhibitory action of marine algae extracts on the Trypanosoma cruzi dihydroorotate dehydrogenase activity and on the protozoan growth in mammalian cells (2005)
  • Author(s): Nara, T., Kamei, Y., Tsubouchi, A., Annoura, T., Hirota, K., Iizumi, K., Dohmoto, Y, Ono, T., Aoki, T
  • Journal Title: Parasitollnt 54 Pages: 59-64
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Expression, purification and crystallization of Trypanosoma cruzi dihydroorotate dehydrogenase complexed with orotate (2005)
  • Author(s): Inaoka, DK., Takashima, E., Osanai, A., Shimizu, H., Nara, T., Aoki, T., Harada, S., Kita, K
  • Journal Title: Acta Cryst F61 Pages: 875-878
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Antichagasic activity of komaroviquinone is due to generation of reactive oxygen species catalyzed by Trypanosoma cruzi old yellow enzyme (2005)
  • Author(s): Uchiyama, N., Kabututu, Z., Kubata, BK., Kiuchi, F., Ito, M., Nakajima-Shimada, J., Aoki, T., Ohkubo, K., Fukuzumi, S., Martin, SK., Honda, G., Urade, Y
  • Journal Title: Antimicrob Agents Chemother 49 Pages: 5123-5126
  • Description: 「研究成果報告書概要(欧文)」より
• [Book] ハリソン内科学(原著第16版), Part 6感染症, Section 17〜19(第192〜204章)(監訳)福井次矢・黒川清(日本語版監修) (2006)
  • Author(s): 青木 孝
  • Total Pages: 83
  • Publisher: メディカル・サイエンス・インターナショナル,東京
  • Description: 「研究成果報告書概要(和文)」より
• [Remarks] 「研究成果報告書概要(和文)」より
  • URL: http://www.juntendo.ac.jp/graduate/laboratory/labo/kiseityu/k6.html