A screening campaign in sea urchin egg homogenate as a platform for discovering modulators of NAADP-dependent Ca2+ signaling in human cells

Gihan S. Gunaratne; Malcolm E. Johns; Hallie M. Hintz; Timothy F Walseth; Jonathan S. Marchant

doi:10.1016/j.ceca.2018.08.002

A screening campaign in sea urchin egg homogenate as a platform for discovering modulators of NAADP-dependent Ca²⁺ signaling in human cells

Gihan S. Gunaratne, Malcolm E. Johns, Hallie M. Hintz, Timothy F Walseth, Jonathan S. Marchant

Research output: Contribution to journal › Article › peer-review

24 Scopus citations

Abstract

The Ca²⁺ mobilizing second messenger nicotinic acid adenine dinucleotide phosphate (NAADP) regulates intracellular trafficking events, including translocation of certain enveloped viruses through the endolysosomal system. Targeting NAADP-evoked Ca²⁺ signaling may therefore be an effective strategy for discovering novel antivirals as well as therapeutics for other disorders. To aid discovery of novel scaffolds that modulate NAADP-evoked Ca²⁺ signaling in human cells, we have investigated the potential of using the sea urchin egg homogenate system for a screening campaign. Known pharmacological inhibitors of NAADP-evoked Ca²⁺ release (but not cADPR- or IP₃-evoked Ca²⁺ release) in this invertebrate system strongly correlated with inhibition of MERS-pseudovirus infectivity in a human cell line. A primary screen of 1534 compounds yielded eighteen ‘hits’ exhibiting >80% inhibition of NAADP-evoked Ca²⁺ release. A validation pipeline for these candidates yielded seven drugs that inhibited NAADP-evoked Ca²⁺ release without depleting acidic Ca²⁺ stores in a human cell line. These candidates displayed a similar penetrance of inhibition in both the sea urchin system and the human cell line, and the extent of inhibition of NAADP-evoked Ca²⁺ signals correlated well with observed inhibition of infectivity of a Middle East Respiratory syndrome coronavirus (MERS-CoV) pseudovirus. These experiments support the potential of this simple, homogenate system for screening campaigns to discover modulators of NAADP, cADPR and IP₃-dependent Ca²⁺ signaling with potential therapeutic value.

Original language	English (US)
Pages (from-to)	42-52
Number of pages	11
Journal	Cell Calcium
Volume	75
DOIs	https://doi.org/10.1016/j.ceca.2018.08.002
State	Published - Nov 2018

Bibliographical note

Funding Information:
GSG, MEJ, HMH and TFW performed experiments. GG and TFW analyzed data. GG, TFW and JSM collaborated to design experiments. GG and JSM wrote the paper. All authors reviewed the results, and commented upon the final version of the manuscript. Work in the Marchant Lab is support by NIH ( R01 GM088790 ) and Regenerative Medicine Minnesota ( RMM 11215 DS003 ).

Publisher Copyright:
© 2018 Elsevier Ltd

Keywords

Carelease
Drug screening
Endosomes
Lysosomes
NAADP

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Publisher link

10.1016/j.ceca.2018.08.002

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6286156

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title = "A screening campaign in sea urchin egg homogenate as a platform for discovering modulators of NAADP-dependent Ca2+ signaling in human cells",

abstract = "The Ca2+ mobilizing second messenger nicotinic acid adenine dinucleotide phosphate (NAADP) regulates intracellular trafficking events, including translocation of certain enveloped viruses through the endolysosomal system. Targeting NAADP-evoked Ca2+ signaling may therefore be an effective strategy for discovering novel antivirals as well as therapeutics for other disorders. To aid discovery of novel scaffolds that modulate NAADP-evoked Ca2+ signaling in human cells, we have investigated the potential of using the sea urchin egg homogenate system for a screening campaign. Known pharmacological inhibitors of NAADP-evoked Ca2+ release (but not cADPR- or IP3-evoked Ca2+ release) in this invertebrate system strongly correlated with inhibition of MERS-pseudovirus infectivity in a human cell line. A primary screen of 1534 compounds yielded eighteen {\textquoteleft}hits{\textquoteright} exhibiting >80% inhibition of NAADP-evoked Ca2+ release. A validation pipeline for these candidates yielded seven drugs that inhibited NAADP-evoked Ca2+ release without depleting acidic Ca2+ stores in a human cell line. These candidates displayed a similar penetrance of inhibition in both the sea urchin system and the human cell line, and the extent of inhibition of NAADP-evoked Ca2+ signals correlated well with observed inhibition of infectivity of a Middle East Respiratory syndrome coronavirus (MERS-CoV) pseudovirus. These experiments support the potential of this simple, homogenate system for screening campaigns to discover modulators of NAADP, cADPR and IP3-dependent Ca2+ signaling with potential therapeutic value.",

keywords = "Carelease, Drug screening, Endosomes, Lysosomes, NAADP",

author = "Gunaratne, {Gihan S.} and Johns, {Malcolm E.} and Hintz, {Hallie M.} and Walseth, {Timothy F} and Marchant, {Jonathan S.}",

note = "Funding Information: GSG, MEJ, HMH and TFW performed experiments. GG and TFW analyzed data. GG, TFW and JSM collaborated to design experiments. GG and JSM wrote the paper. All authors reviewed the results, and commented upon the final version of the manuscript. Work in the Marchant Lab is support by NIH ( R01 GM088790 ) and Regenerative Medicine Minnesota ( RMM 11215 DS003 ). Publisher Copyright: {\textcopyright} 2018 Elsevier Ltd",

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