To further investigate the importance of insulin signaling in the growth, development, sexual maturation and egg production of adult schistosomes, we have focused attention on the insulin receptors (SjIRs) of Schistosoma japonicum, which we have previously cloned and partially characterised. We now show, by Biolayer Interferometry, that human insulin can bind the L1 subdomain (insulin binding domain) of recombinant (r)SjIR1 and rSjIR2 (designated SjLD1 and SjLD2) produced using the Drosophila S2 protein expression system. We have then used RNA interference (RNAi) to knock down the expression of the SjIRs in adult S. japonicum in vitro and show that, in addition to their reduced transcription, the transcript levels of other important downstream genes within the insulin pathway, associated with glucose metabolism and schistosome fecundity, were also impacted substantially. Further, a significant decrease in glucose uptake was observed in the SjIR-knockdown worms compared with luciferase controls. In vaccine/challenge experiments, we found that rSjLD1 and rSjLD2 depressed female growth, intestinal granuloma density and faecal egg production in S. japonicum in mice presented with a low dose challenge infection. These data re-emphasize the potential of the SjIRs as veterinary transmission blocking vaccine candidates against zoonotic schistosomiasis japonica in China and the Philippines.
References
[1]
Wilson MS, Mentink-Kane MM, Pesce JT, Ramalingam TR, Thompson R, et al. Immunopathology of schistosomiasis. Immunol Cell Biol 2007; 85: 148–154. pmid:17160074 doi: 10.1038/sj.icb.7100014
[2]
Ross AG, Bartley PB, Sleigh AC, Olds GR, Li Y, et al. Schistosomiasis. N Engl J Med 2002; 346: 1212–1220. pmid:11961151 doi: 10.1056/nejmra012396
[3]
Gonnert R, Andrews P Praziquantel, a new board-spectrum antischistosomal agent. Z Parasitenkd 1977; 52: 129–150. pmid:410178 doi: 10.1007/bf00389899
[4]
McManus DP, Gray DJ, Li Y, Feng Z, Williams GM, et al. Schistosomiasis in the People's Republic of China: the era of the Three Gorges Dam. Clin Microbiol Rev 2010; 23: 442–466. doi: 10.1128/CMR.00044-09. pmid:20375361
[5]
Gordon CA, Acosta LP, Gray DJ, Olveda RM, Jarilla B, et al. High prevalence of Schistosoma japonicum infection in Carabao from Samar Province, the Philippines: implications for transmission and control. PLoS Negl Trop Dis 2012; 6: e1778. doi: 10.1371/journal.pntd.0001778. pmid:23029571
[6]
McManus DP, Loukas A Current status of vaccines for schistosomiasis. Clin Microbiol Rev 2008; 21: 225–242. doi: 10.1128/CMR.00046-07. pmid:18202444
[7]
Da'dara AA, Li YS, Xiong T, Zhou J, Williams GM, et al. DNA-based vaccines protect against zoonotic schistosomiasis in water buffalo. Vaccine 2008; 26: 3617–3625. doi: 10.1016/j.vaccine.2008.04.080. pmid:18524429
[8]
Mo AX, Agosti JM, Walson JL, Hall BF, Gordon L Schistosomiasis elimination strategies and potential role of a vaccine in achieving global health goals. Am J Trop Med Hyg 2014; 90: 54–60. doi: 10.4269/ajtmh.13-0467. pmid:24402703
[9]
Morel M, Vanderstraete M, Hahnel S, Grevelding CG, Dissous C Receptor tyrosine kinases and schistosome reproduction: new targets for chemotherapy. Front Genet 2014; 5: 238. doi: 10.3389/fgene.2014.00238. pmid:25101117
[10]
Bueding E Carbohydrate metabolism of Schistosoma mansoni. J Gen Physiol 1950; 33: 475–495. pmid:15422103 doi: 10.1085/jgp.33.5.475
[11]
Krautz-Peterson G, Simoes M, Faghiri Z, Ndegwa D, Oliveira G, et al. Suppressing glucose transporter gene expression in schistosomes impairs parasite feeding and decreases survival in the mammalian host. PLoS Pathog 2010; 6: e1000932. doi: 10.1371/journal.ppat.1000932. pmid:20532163
[12]
Khayath N, Vicogne J, Ahier A, BenYounes A, Konrad C, et al. Diversification of the insulin receptor family in the helminth parasite Schistosoma mansoni. Febs J 2007; 274: 659–676. pmid:17181541 doi: 10.1111/j.1742-4658.2006.05610.x
[13]
You H, Zhang W, Jones MK, Gobert GN, Mulvenna J, et al. Cloning and characterisation of Schistosoma japonicum insulin receptors. PLoS One 2010; 5: e9868. doi: 10.1371/journal.pone.0009868. pmid:20352052
[14]
Feng Y, Williams BG, Koumanov F, Wolstenholme AJ, Holman GD FGT-1 is the major glucose transporter in C. elegans and is central to aging pathways. Biochem J 2013; 456: 219–229. doi: 10.1042/BJ20131101. pmid:24024580
[15]
You H, Zhang W, Moertel L, McManus DP, Gobert GN Transcriptional profiles of adult male and female Schistosoma japonicum in response to insulin reveal increased expression of genes involved in growth and development. Int J Parasitol 2009; 39: 1551–1559. doi: 10.1016/j.ijpara.2009.06.006. pmid:19596015
[16]
Vanderstraete M, Gouignard N, Cailliau K, Morel M, Hahnel S, et al. Venus kinase receptors control reproduction in the platyhelminth parasite Schistosoma mansoni. PLoS Pathog 2014; 10: e1004138. doi: 10.1371/journal.ppat.1004138. pmid:24875530
[17]
You H, Gobert GN, Duke MG, Zhang W, Li Y, et al. The insulin receptor is a transmission blocking veterinary vaccine target for zoonotic Schistosoma japonicum. Int J Parasitol 2012; 42: 801–807. doi: 10.1016/j.ijpara.2012.06.002. pmid:22771861
[18]
Gobert GN, Moertel L, Brindley PJ, McManus DP Developmental gene expression profiles of the human pathogen Schistosoma japonicum. BMC Genomics 2009; 10: 128. doi: 10.1186/1471-2164-10-128. pmid:19320991
[19]
Morales ME, Rinaldi G, Gobert GN, Kines KJ, Tort JF, et al. RNA interference of Schistosoma mansoni cathepsin D, the apical enzyme of the hemoglobin proteolysis cascade. Mol Biochem Parasitol 2008; 157: 160–168. pmid:18067980 doi: 10.1016/j.molbiopara.2007.10.009
[20]
Rinaldi G, Morales ME, Alrefaei YN, Cancela M, Castillo E, et al. RNA interference targeting leucine aminopeptidase blocks hatching of Schistosoma mansoni eggs. Mol Biochem Parasitol 2009; 167: 118–126. doi: 10.1016/j.molbiopara.2009.05.002. pmid:19463860
[21]
You H, McManus DP, Hu W, Smout MJ, Brindley PJ, et al. Transcriptional responses of in vivo praziquantel exposure in schistosomes identifies a functional role for calcium signalling pathway member CamKII. PLoS Pathog 2013; 9: e1003254. doi: 10.1371/journal.ppat.1003254. pmid:23555262
[22]
Kasinathan RS, Sharma LK, Cunningham C, Webb TR, Greenberg RM Inhibition or knockdown of ABC transporters enhances susceptibility of adult and juvenile schistosomes to Praziquantel. PLoS Negl Trop Dis 2014; 8: e3265. doi: 10.1371/journal.pntd.0003265. pmid:25330312
[23]
Geyer KK, Rodriguez Lopez CM, Chalmers IW, Munshi SE, Truscott M, et al. Cytosine methylation regulates oviposition in the pathogenic blood fluke Schistosoma mansoni. Nat Commun 2011; 2: 424. doi: 10.1038/ncomms1433. pmid:21829186
[24]
Pearce EJ, James SL, Dalton J, Barrall A, Ramos C, et al. Immunochemical characterization and purification of Sm-97, a Schistosoma mansoni antigen monospecifically recognized by antibodies from mice protectively immunized with a nonliving vaccine. J Immunol 1986; 137: 3593–3600. pmid:2431037
[25]
Zhou S, Liu S, Song G, Xu Y [Cloning, sequencing and expression of the full-length gene encoding paramyosin of Schistosoma japonicum in vivo]. Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi 1999; 17: 196–199. pmid:12563755
[26]
Abdiche Y, Malashock D, Pinkerton A, Pons J Determining kinetics and affinities of protein interactions using a parallel real-time label-free biosensor, the Octet. Anal Biochem 2008; 377: 209–217. doi: 10.1016/j.ab.2008.03.035. pmid:18405656
[27]
Nakamura S, Sakata S, Atassi MZ Localization and synthesis of an insulin-binding region on human insulin receptor. J Protein Chem 1990; 9: 229–233. pmid:2201312 doi: 10.1007/bf01025313
[28]
Golubovskaya VM, Ho B, Zheng M, Magis A, Ostrov D, et al. Disruption of focal adhesion kinase and p53 interaction with small molecule compound R2 reactivated p53 and blocked tumor growth. BMC Cancer 2013; 13: 342. doi: 10.1186/1471-2407-13-342. pmid:23841915
[29]
McKern NM, Lawrence MC, Streltsov VA, Lou MZ, Adams TE, et al. Structure of the insulin receptor ectodomain reveals a folded-over conformation. Nature 2006; 443: 218–221. pmid:16957736 doi: 10.1038/nature05106
[30]
Schistosoma japonicum Genome Sequencing and Functional Analysis Consortium.The Schistosoma japonicum genome reveals features of host-parasite interplay. Nature 2009; 460: 345–351. doi: 10.1038/nature08140. pmid:19606140
[31]
Okada T, Kawano Y, Sakakibara T, Hazeki O, Ui M Essential role of phosphatidylinositol 3-kinase in insulin-induced glucose transport and antilipolysis in rat adipocytes. Studies with a selective inhibitor wortmannin. J Biol Chem 1994; 269: 3568–3573. pmid:8106400
[32]
Chang L, Chiang SH, Saltiel AR Insulin signaling and the regulation of glucose transport. Mol Med 2004; 10: 65–71. pmid:16307172
[33]
Hemer S, Konrad C, Spiliotis M, Koziol U, Schaack D, et al. Host insulin stimulates Echinococcus multilocularis insulin signalling pathways and larval development. BMC Biol 2014; 12: 5. doi: 10.1186/1741-7007-12-5. pmid:24468049
[34]
Nielsen JN, Wojtaszewski JF Regulation of glycogen synthase activity and phosphorylation by exercise. Proc Nutr Soc 2004; 63: 233–237. pmid:15294036 doi: 10.1079/pns2004348
[35]
Ahier A, Khayath N, Vicogne J, Dissous C Insulin receptors and glucose uptake in the human parasite Schistosoma mansoni. Parasite 2008; 15: 573–579. pmid:19202764 doi: 10.1051/parasite/2008154573
[36]
Vanderstraete M, Gouignard N, Cailliau K, Morel M, Lancelot J, et al. Dual Targeting of Insulin and Venus Kinase Receptors of Schistosoma mansoni for Novel Anti-schistosome Therapy. PLoS Negl Trop Dis 2013; 7: e2226. doi: 10.1371/journal.pntd.0002226. pmid:23696913
[37]
Sasaoka T, Kobayashi M The functional significance of Shc in insulin signaling as a substrate of the insulin receptor. Endocr J 2000; 47: 373–381. pmid:11075717 doi: 10.1507/endocrj.47.373
[38]
Kanzaki M, Pessin JE Insulin signaling: GLUT4 vesicles exit via the exocyst. Curr Biol 2003; 13: R574–576. pmid:12867054 doi: 10.1016/s0960-9822(03)00478-0
[39]
Camacho M, Agnew A Schistosoma: rate of glucose import is altered by acetylcholine interaction with tegumental acetylcholine receptors and acetylcholinesterase. Exp Parasitol 1995; 81: 584–591. pmid:8543000 doi: 10.1006/expr.1995.1152
[40]
Chen JH, Wang H, Chen JX, Bergquist R, Tanner M, et al. Frontiers of parasitology research in the People's Republic of China: infection, diagnosis, protection and surveillance. Parasit Vectors 2012; 5: 221. doi: 10.1186/1756-3305-5-221. pmid:23036110
[41]
Pearce EJ Parasitology: Rejuvenation through stem cells. Nature 2013; 494: 438–439. doi: 10.1038/nature11953. pmid:23426259
Argiro L, Henri S, Dessein H, Kouriba B, Dessein AJ, et al. Induction of a protection against S. mansoni with a MAP containing epitopes of Sm37-GAPDH and Sm10-DLC. Effect of coadsorption with GM-CSF on alum. Vaccine 2000; 18: 2033–2038. pmid:10706966 doi: 10.1016/s0264-410x(99)00523-x
