Snake Venom Gland Organoids

Yorick Post; Jens Puschhof; Joep Beumer; Harald M. Kerkkamp; Merijn A.G. de Bakker; Julien Slagboom; Buys de Barbanson; Nienke R. Wevers; Xandor M. Spijkers; Thomas Olivier; Taline D. Kazandjian; Stuart Ainsworth; Carmen Lopez Iglesias; Willine J. van de Wetering; Maria C. Heinz; Ravian L. van Ineveld; Regina G.D.M. van Kleef; Harry Begthel; Jeroen Korving; Yotam E. Bar-Ephraim; Walter Getreuer; Anne C. Rios; Remco H.S. Westerink; Hugo J.G. Snippert; Alexander van Oudenaarden; Peter J. Peters; Freek J. Vonk; Jeroen Kool; Michael K. Richardson; Nicholas R. Casewell; Hans Clevers

doi:10.1016/j.cell.2019.11.038

Snake Venom Gland Organoids

Yorick Post, Jens Puschhof, Joep Beumer, Harald M. Kerkkamp, Merijn A.G. de Bakker, Julien Slagboom, Buys de Barbanson, Nienke R. Wevers, Xandor M. Spijkers, Thomas Olivier, Taline D. Kazandjian, Stuart Ainsworth, Carmen Lopez Iglesias, Willine J. van de Wetering, Maria C. Heinz, Ravian L. van Ineveld, Regina G.D.M. van Kleef, Harry Begthel, Jeroen Korving, Yotam E. Bar-EphraimWalter Getreuer, Anne C. Rios, Remco H.S. Westerink, Hugo J.G. Snippert, Alexander van Oudenaarden, Peter J. Peters, Freek J. Vonk, Jeroen Kool, Michael K. Richardson, Nicholas R. Casewell, Hans Clevers^*

^*Corresponding author for this work

AIMMS

Research output: Contribution to Journal › Article › Academic › peer-review

98 Downloads (Pure)

Abstract

Wnt dependency and Lgr5 expression define multiple mammalian epithelial stem cell types. Under defined growth factor conditions, such adult stem cells (ASCs) grow as 3D organoids that recapitulate essential features of the pertinent epithelium. Here, we establish long-term expanding venom gland organoids from several snake species. The newly assembled transcriptome of the Cape coral snake reveals that organoids express high levels of toxin transcripts. Single-cell RNA sequencing of both organoids and primary tissue identifies distinct venom-expressing cell types as well as proliferative cells expressing homologs of known mammalian stem cell markers. A hard-wired regional heterogeneity in the expression of individual venom components is maintained in organoid cultures. Harvested venom peptides reflect crude venom composition and display biological activity. This study extends organoid technology to reptilian tissues and describes an experimentally tractable model system representing the snake venom gland.

Original language	English
Pages (from-to)	233-247.e21
Number of pages	37
Journal	Cell
Volume	180
Issue number	2
Early online date	23 Jan 2020
DOIs	https://doi.org/10.1016/j.cell.2019.11.038
Publication status	Published - 23 Jan 2020

Funding

We thank Reinier van der Linden for flow cytometry assistance; Benedetta Artegiani and Delilah Hendriks for generation of CRISPR-HOT technology and assistance in applying it to snake venom gland organoids; Anko de Graaff and the Hubrecht Imaging Centre (HIC) for microscopy assistance; BaseClear B.V. for bulk mRNA sequencing and de novo transcriptome assembly; Bas Ponsioen for providing the lentiviral H2B-RFP construct; Single Cell Discoveries for the provided single-cell sequencing service and support; Jeremie Tai-A-Pin and Harold van der Ploeg for donating venom gland material to this study; and Sebastiaan Voskuil, Edwin Boel, Marc Bonten, and Frank Driehuis for advice regarding antibiotic treatments. X.M.S. was supported by ALS Foundation Netherlands . N.R.C. was supported by a Sir Henry Dale Fellowship ( 200517/Z/16/Z ) jointly funded by the Wellcome Trust and the Royal Society .

Funders	Funder number
Edwin Boel
Hubrecht Imaging Centre
Mimetas
Sebastiaan Voskuil
Wellcome Trust
Royal Society
Stichting ALS Nederland	200517/Z/16/Z

Keywords

heterogeneity
knock-in reporter
Lgr5
organoid
single cell RNA sequencing
snake
stem cells
transcriptomics
venom gland

UN SDGs

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

Access to Document

10.1016/j.cell.2019.11.038

Snake Venom Gland OrganoidsFinal published version, 10.5 MBLicence: Article 25fa Dutch Copyright Act

Persistent URL (handle)

Persistent link

Cite this

Post, Y., Puschhof, J., Beumer, J., Kerkkamp, H. M., de Bakker, M. A. G., Slagboom, J., de Barbanson, B., Wevers, N. R., Spijkers, X. M., Olivier, T., Kazandjian, T. D., Ainsworth, S., Iglesias, C. L., van de Wetering, W. J., Heinz, M. C., van Ineveld, R. L., van Kleef, R. G. D. M., Begthel, H., Korving, J., ... Clevers, H. (2020). Snake Venom Gland Organoids. Cell, 180(2), 233-247.e21. https://doi.org/10.1016/j.cell.2019.11.038

@article{ff5336a23e024d9c89972e4396580d89,

title = "Snake Venom Gland Organoids",

abstract = "Wnt dependency and Lgr5 expression define multiple mammalian epithelial stem cell types. Under defined growth factor conditions, such adult stem cells (ASCs) grow as 3D organoids that recapitulate essential features of the pertinent epithelium. Here, we establish long-term expanding venom gland organoids from several snake species. The newly assembled transcriptome of the Cape coral snake reveals that organoids express high levels of toxin transcripts. Single-cell RNA sequencing of both organoids and primary tissue identifies distinct venom-expressing cell types as well as proliferative cells expressing homologs of known mammalian stem cell markers. A hard-wired regional heterogeneity in the expression of individual venom components is maintained in organoid cultures. Harvested venom peptides reflect crude venom composition and display biological activity. This study extends organoid technology to reptilian tissues and describes an experimentally tractable model system representing the snake venom gland.",

keywords = "heterogeneity, knock-in reporter, Lgr5, organoid, single cell RNA sequencing, snake, stem cells, transcriptomics, venom gland",

author = "Yorick Post and Jens Puschhof and Joep Beumer and Kerkkamp, \{Harald M.\} and \{de Bakker\}, \{Merijn A.G.\} and Julien Slagboom and \{de Barbanson\}, Buys and Wevers, \{Nienke R.\} and Spijkers, \{Xandor M.\} and Thomas Olivier and Kazandjian, \{Taline D.\} and Stuart Ainsworth and Iglesias, \{Carmen Lopez\} and \{van de Wetering\}, \{Willine J.\} and Heinz, \{Maria C.\} and \{van Ineveld\}, \{Ravian L.\} and \{van Kleef\}, \{Regina G.D.M.\} and Harry Begthel and Jeroen Korving and Bar-Ephraim, \{Yotam E.\} and Walter Getreuer and Rios, \{Anne C.\} and Westerink, \{Remco H.S.\} and Snippert, \{Hugo J.G.\} and \{van Oudenaarden\}, Alexander and Peters, \{Peter J.\} and Vonk, \{Freek J.\} and Jeroen Kool and Richardson, \{Michael K.\} and Casewell, \{Nicholas R.\} and Hans Clevers",

year = "2020",

month = jan,

day = "23",

doi = "10.1016/j.cell.2019.11.038",

language = "English",

volume = "180",

pages = "233--247.e21",

journal = "Cell",

issn = "0092-8674",

publisher = "Elsevier B.V.",

number = "2",

}

Post, Y, Puschhof, J, Beumer, J, Kerkkamp, HM, de Bakker, MAG, Slagboom, J, de Barbanson, B, Wevers, NR, Spijkers, XM, Olivier, T, Kazandjian, TD, Ainsworth, S, Iglesias, CL, van de Wetering, WJ, Heinz, MC, van Ineveld, RL, van Kleef, RGDM, Begthel, H, Korving, J, Bar-Ephraim, YE, Getreuer, W, Rios, AC, Westerink, RHS, Snippert, HJG, van Oudenaarden, A, Peters, PJ, Vonk, FJ, Kool, J, Richardson, MK, Casewell, NR & Clevers, H 2020, 'Snake Venom Gland Organoids', Cell, vol. 180, no. 2, pp. 233-247.e21. https://doi.org/10.1016/j.cell.2019.11.038

TY - JOUR

T1 - Snake Venom Gland Organoids

AU - Post, Yorick

AU - Puschhof, Jens

AU - Beumer, Joep

AU - Kerkkamp, Harald M.

AU - de Bakker, Merijn A.G.

AU - Slagboom, Julien

AU - de Barbanson, Buys

AU - Wevers, Nienke R.

AU - Spijkers, Xandor M.

AU - Olivier, Thomas

AU - Kazandjian, Taline D.

AU - Ainsworth, Stuart

AU - Iglesias, Carmen Lopez

AU - van de Wetering, Willine J.

AU - Heinz, Maria C.

AU - van Ineveld, Ravian L.

AU - van Kleef, Regina G.D.M.

AU - Begthel, Harry

AU - Korving, Jeroen

AU - Bar-Ephraim, Yotam E.

AU - Getreuer, Walter

AU - Rios, Anne C.

AU - Westerink, Remco H.S.

AU - Snippert, Hugo J.G.

AU - van Oudenaarden, Alexander

AU - Peters, Peter J.

AU - Vonk, Freek J.

AU - Kool, Jeroen

AU - Richardson, Michael K.

AU - Casewell, Nicholas R.

AU - Clevers, Hans

PY - 2020/1/23

Y1 - 2020/1/23

N2 - Wnt dependency and Lgr5 expression define multiple mammalian epithelial stem cell types. Under defined growth factor conditions, such adult stem cells (ASCs) grow as 3D organoids that recapitulate essential features of the pertinent epithelium. Here, we establish long-term expanding venom gland organoids from several snake species. The newly assembled transcriptome of the Cape coral snake reveals that organoids express high levels of toxin transcripts. Single-cell RNA sequencing of both organoids and primary tissue identifies distinct venom-expressing cell types as well as proliferative cells expressing homologs of known mammalian stem cell markers. A hard-wired regional heterogeneity in the expression of individual venom components is maintained in organoid cultures. Harvested venom peptides reflect crude venom composition and display biological activity. This study extends organoid technology to reptilian tissues and describes an experimentally tractable model system representing the snake venom gland.

AB - Wnt dependency and Lgr5 expression define multiple mammalian epithelial stem cell types. Under defined growth factor conditions, such adult stem cells (ASCs) grow as 3D organoids that recapitulate essential features of the pertinent epithelium. Here, we establish long-term expanding venom gland organoids from several snake species. The newly assembled transcriptome of the Cape coral snake reveals that organoids express high levels of toxin transcripts. Single-cell RNA sequencing of both organoids and primary tissue identifies distinct venom-expressing cell types as well as proliferative cells expressing homologs of known mammalian stem cell markers. A hard-wired regional heterogeneity in the expression of individual venom components is maintained in organoid cultures. Harvested venom peptides reflect crude venom composition and display biological activity. This study extends organoid technology to reptilian tissues and describes an experimentally tractable model system representing the snake venom gland.

KW - heterogeneity

KW - knock-in reporter

KW - Lgr5

KW - organoid

KW - single cell RNA sequencing

KW - snake

KW - stem cells

KW - transcriptomics

KW - venom gland

UR - https://www.scopus.com/pages/publications/85077840511

UR - https://www.scopus.com/inward/citedby.url?scp=85077840511&partnerID=8YFLogxK

U2 - 10.1016/j.cell.2019.11.038

DO - 10.1016/j.cell.2019.11.038

M3 - Article

AN - SCOPUS:85077840511

SN - 0092-8674

VL - 180

SP - 233-247.e21

JO - Cell

JF - Cell

IS - 2

ER -

Snake Venom Gland Organoids

Abstract

Funding

Keywords

UN SDGs

Access to Document

Persistent URL (handle)

Other files and links

Fingerprint

Cite this