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Pulchelloid A, a sesquiterpene lactone from the Canadian prairie plant Gaillardia aristata inhibits mitosis in human cells

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Abstract

Background

The Canadian prairie ecosystem presents a rich source of natural products from plants that are subjected to herbivory by grazing mammals. This type of ecological competition may contribute to the production of natural products of interest in cell biology and medical research. We provide the first biological description of the sesquiterpene lactone, pulchelloid A, which we isolated from the prairie plant, Gaillardia aristata (Asteraceae) and report that it inhibits mitosis in human cells.

Methods and results

We found that G. aristata (Blanket flower) extracts were cytotoxic to human cell lines and used phenotypic assays to characterize the bioactivity of extracts. Before dying, cells were characterized by a rounded morphology, phospho-histone H3 signals, mitotic spindles, and active Cdk1. By biology-guided fractionation of Gaillardia extracts, we isolated a sesquiterpene lactone named pulchelloid A. We used immunofluorescence microscopy and observed that cells treated with pulchelloid A have phospho-histone H3 positive chromosomes and a mitotic spindle, confirming that they were in mitosis. Treated cells arrest with an unusual phenotype; they enter a prolonged mitotic arrest in which the spindles become multipolar and the chromosomes acquire histone γH2AX foci, a hallmark of damaged DNA.

Conclusions

We propose that pulchelloid A, a natural product present in the prairie plant Gaillardia aristata, delays cells in mitosis. There is a growing body of evidence that a small number of members of the sesquiterpene lactone chemical family may target proteins that regulate mitosis.

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Data Availability

All data and materials support their published claims and comply with field standards.

Abbreviations

6-OAP:

6-O-angeloylplenolin

Cdk1:

Cyclin dependent kinase 1

CPT:

Camptothecin

DAPI:

4′,6-Diamidino-2-phenylindole

DMSO:

Dimethylsulfoxide

F.E.:

Flower extract

h:

Hours

L.E.:

Leaf extract

NT:

Not-treated

Noco:

Nocodazole

PA:

Pulchelloid A

PH3:

Phospho-histone H3

S.E.:

Stem extract

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Acknowledgements

We thank Joanne Golden, Dr. John Bain, Jan Tuescher and members of the Natural Product and Cancer Cell Laboratories for valuable discussions. We thank the following organizations and people for granting permission to collect plants on public or private lands: Alberta Environment and Parks, Alberta Tourism Parks and Recreation, City of Lethbridge, and Mr. and Mrs. Morden, Mr. Michael Monaghan. The following people kindly volunteered in plant collections: Chad Beck, Chloe Devoy, Audrey Golsteyn, Charlène Golsteyn, Quentin Golsteyn, Paul Landry, Randy Paskuski.

Funding

Funding for this project was provided by Canada Foundation for Innovation [#34542], Alberta Innovates Sustainability Fund, the University of Lethbridge Office of Research and Innovation Services, and NSERC Discovery Grants [#RGPIN-2017–04398], as well as the Swiss National Center for Competence (NCCR) in Chemical Biology funded by the Swiss National Science Foundation.

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Authors and Affiliations

  1. Natural Product Laboratory, University of Lethbridge, Lethbridge, AB, Canada

    Alessandra Bosco, Layla Molina & Roy M. Golsteyn

  2. Microbial Research Group, Lethbridge College, Lethbridge, AB, Canada

    Sophie M. Kernéis

  3. Swiss Federal Institute of Technology Lausanne (EPFL), Lausanne, Switzerland

    Georgios Hatzopoulos, Tatiana Favez & Pierre Gönczy

  4. Department of Earth, Ocean, Atmospheric Sciences, University of British Columbia, Vancouver, BC, Canada

    Cholpisut Tantapakul, Wisanu Maneerat, Benjamin Yeremy, David E. Williams & Raymond J. Andersen

Authors
  1. Alessandra Bosco
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  2. Layla Molina
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  3. Sophie M. Kernéis
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Contributions

AB, LM, SMK, RMG harvested plants, prepared extracts, and performed experiments. CT, WM, BY, DEW participated in purification and structure determination of pulchelloid AGH, TF performed experiments to analyze mitotic spindles, centrosomes, and centrioles. AB, LM, SMK, PG, RJA and RMG participated in experimental design, data analysis and wrote the manuscript.

Corresponding author

Correspondence to Roy M. Golsteyn.

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Bosco, A., Molina, L., Kernéis, S.M. et al. Pulchelloid A, a sesquiterpene lactone from the Canadian prairie plant Gaillardia aristata inhibits mitosis in human cells. Mol Biol Rep 48, 5459–5471 (2021). https://doi.org/10.1007/s11033-021-06554-z

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