Study of the role of P-Glycoprotein-2 of the parasitic nematode <em>Heligmosomoides polygyrus</em> in the cholesterol uptake

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Study of the role of P-Glycoprotein-2 of the parasitic nematode Heligmosomoides polygyrus in the cholesterol

uptake

Mohamed Issouf, Fatima Mohamed, Cédric Neveu

To cite this version:

Mohamed Issouf, Fatima Mohamed, Cédric Neveu. Study of the role of P-Glycoprotein-2 of the parasitic nematode Heligmosomoides polygyrus in the cholesterol uptake. 27. Conference of the World Association for the Advancement of Veterinary Parasitology, Jul 2019, Madison, United States.

349 p., 2019, Abstract book - 27. WAAVP. �hal-02736800�

WAAVP2019 ^

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Conference of the World Association for the Advancements of Veterinary Parasitology 1

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American Association of Veterinary Parasitologists Annual Meeting 63

Annual Livestock Insect Workers Conference

Sifting and Winnowing the Evidence in Veterinary Parasitology

27 ^th Conference of the World Association for the Advancement of Veterinary Parasitology

JULY 7 – 11, 2019 | MADISON, WI, USA

Dedicated to the legacy of Professor Arlie C. Todd

Abstract Book

#WAAVP2019 www.WAAVP2019.com

Joint meeting with the 64 ^th American Association of Veterinary Parasitologists Annual Meeting & the 63 ^rd Annual Livestock Insect Workers Conference

@WAAVP_2019

@WAAVP2019

WAAVP2019 ^

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Conference of the World Association for the Advancements of Veterinary Parasitology 198

64

American Association of Veterinary Parasitologists Annual Meeting 63

Annual Livestock Insect Workers Conference

PS01.18 Exploring Thermosensory Behaviours and Pathways in Filarial Nematode Parasites Mr. Zachary Heimark

, Mrs. Katherine Kudrna

, Dr. Nicolas Wheeler

, Dr. Mostafa Zamanian

1

University Of Wisconsin-Madison, United States Dirofilaria immitis is the etiologic agent of canine and feline heartworm disease.

The prevention of this disease is achieved through the administration of macrocyclic lactones as a prophylactic, or melarsomine dihydrochloride in dogs with adult infections.

Severely limited pharmacologic options make control of this disease difficult, especially in regions of drug resistance or areas with high non-compliance of preventative supplementation. Therefore, in order to decrease the reliance on a single-drug group, other prevention strategies need to be explored. L3 and L4 larval stages of D. immitis are the targets for existing chemotherapies, and these stages exhibit thermosensory behaviours in vitro, which are likely used during transmission and infection of the mammalian host. This behaviour could be exploited by chemotherapeutic intervention, but the molecular effectors of thermosensation are unknown in D. immitis.

To identify putative thermosensory effects in D. immitis, we conducted a pan-phylum comparative genomic and phylogenetic analysis informed by thermosensory

pathways elucidated in the model nematode Caenorhabditis elegans. To validate these putative thermosensory effectors, we developed a reverse genetics approach to knock-down expression of gene targets predicted to function in thermosensation and assayed the modulation of thermosensory phenotypes. Because of the fragmented nature of D. immitis gene predictions, we used long-read isoform sequencing of adult males and females to annotate full- length genes, which enabled the design of improved dsRNA triggers. Larval worms were exposed to dsRNA during in vitro culture or by injecting infected mosquitos, and worms were tested for thermosensory defects using a linear thermal gradient. In addition to RNAi, we also treated larval worms with chemicals that are known to interact with predicted thermosensory effectors and tested for thermosensory defects. We

foresee this approach enabling the continued identification and annotation of D. immitis thermosensory effectors, which could be putative targets for the prevention of heartworm infections.

PS01.19 Study of the Role of

P-Glycoprotein-2 of the Parasitic Nematode Heligmosomoides Polygyrus in the

Cholesterol Uptake

Mohamed Issouf

, Fatima Mohamed

, Cédric Neveu

1

INRA, UR1282 Infectiologie Animale et Santé Publique, Centre de Recherches de Tours, France

Recent studies have described the involvement of Caenorhabditis elegans p-glycoprotein-2 in lysosome formation and lipid storage. In that respect, we are carefully trying to identify in Heligmosomoides

polygyrus bakeri the free-living nematode model Caenorhabtidis elegans p-glycoprotein homologs using degenerate primers. These experiments have allowed the identification of five partial p-glycoprotein sequences. Among those Hba-pgp-2 has particular interest.

Since, C. elegans PGP-2 is involved in the transport of cholesterol. Cholesterol is needed for molting and growth of the nematodes.

In this study, a full-length cDNA sequence corresponding to Hba-pgp-2 was obtained.

It presents high homologies with the Cel- pgp-2 gene sequence. Moreover, we showed that Hba-pgp-2 is expressed among the eggs, fourth-stage larvae and adult and their expression could be induced by cholesterol. In order to characterize the Hba-pgp-2 function, the Hba-pgp-2 promoter was analyzed

using the nematode model C. elegans and complementation of C. elegans pgp-2 mutant by Hba-pgp-2 was performed. This opens the way for studying the involvement of Pgps in parasitic biological process that can lead to develop new way to control nematode parasites.

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