Suppression of Ov-grn-1 encoding granulin of Opisthorchis viverrini inhibits proliferation of biliary epithelial cells

Abstract

Multistep processes likely underlie cholangiocarcinogenesis induced by chronic infection with the fish-borne liver fluke, Opisthorchis viverrini. One process appears to be cellular proliferation of the host bile duct epithelia driven by excretory-secretory (ES) products of this pathogen. Specifically, the secreted growth factor Ov-GRN-1, a liver fluke granulin, is a prominent component of ES and a known driver of hyper-proliferation of cultured human and mouse cells in vitro. We show potent hyper-proliferation of human cholangiocytes induced by low nanomolar levels of recombinant Ov-GRN-1 and similar growth produced by low microgram concentrations of ES products and soluble lysates of the adult worm. To further explore the influence of Ov-GRN-1 on the flukes and the host cells, expression of Ov-grn-1 was repressed using RNA interference. Expression of Ov-grn-1 was suppressed by 95% by day 3 and by ~100% by day 7. Co-culture of Ov-grn-1 suppressed flukes with human cholangiocyte (H-69) or human cholangiocarcinoma (KKU-M214) cell lines retarded cell hyper-proliferation by 25% and 92%, respectively. Intriguingly, flukes in which expression of Ov-grn-1 was repressed were less viable in culture, suggesting that Ov-GRN-1 is an essential growth factor for survival of the adult stage of O. viverrini survival, at least in vitro. To summarize, specific knock down of Ov-grn-1 reduced in vitro survival and capacity of ES products to drive host cell proliferation. These findings may help to contribute to a deeper understanding of liver fluke induced cholangiocarcinogenesis.

Development of a Potent Wound Healing Agent Based on the Liver Fluke Granulin Structural Fold

“…1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 18 The non-malignant cholangiocyte cell line H69 is a SV40-transformed human bile duct epithelial cell line derived from human liver, kindly provided by Dr. Gregory J. Gores, Mayo Clinic, Rochester, Minnesota. H69 cells 23,29,30 were maintained in T75cm 2 vented flasks (Corning) as monolayers as described 31 …”

Biomphalaria glabrata Granulin Increases Resistance to Schistosoma mansoni Infection in Several Biomphalaria Species and Induces the Production of Reactive Oxygen Species by Haemocytes

“…These similarities led us to hypothesize that the effects of BgGRN on the capacity of snails to fight of S. mansoni infections might be conserved across various strains of B. glabrata and various Biomphalaria species due to their evolutionary relatedness. We also had reason to believe that the recombinant granulin we generated from B. glabrata would retain its functionality across Biomphalaria species due to the fact that granulin from Opisthorchis viverrini is capable of inducing cell proliferation in humans, despite the distance between these two species [34,35].…”

Effect of Opisthorchis felineus infection and dimethylnitrosamine administration on the induction of cholangiocarcinoma in Syrian hamsters

“…It is believed that liver flukes can cause damage to the bile duct epithelium in several ways: (i) via mechanical damage; (ii) as a result of action of reactive oxygen species that are produced by the host cells at the site of inflammation; and (iii) because of direct action of fluke-secreted proteins that induce cell proliferation and inhibit apoptosis [34–36]. All these processes lead to lesions in the genetic material and mutations, which get fixed after DNA replication.…”