Analysis of Runx1 Using Induced Gene Ablation Reveals Its Essential Role in Pre-liver HSC Development and Limitations of an In Vivo Approach

Jordi Senserrich; Antoniana Batsivari; Stanislav Rybtsov; Sabrina Gordon-Keylock; Celine Souilhol; Frank Buchholz; David Hills; Suling Zhao; Alexander Medvinsky

doi:10.1016/j.stemcr.2018.08.004

Analysis of Runx1 Using Induced Gene Ablation Reveals Its Essential Role in Pre-liver HSC Development and Limitations of an In Vivo Approach

Jordi Senserrich, Antoniana Batsivari, Stanislav Rybtsov, Sabrina Gordon-Keylock, Celine Souilhol, Frank Buchholz, David Hills, Suling Zhao, Alexander Medvinsky

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

Abstract / Description of output

Hematopoietic stem cells (HSCs) develop in the embryonic aorta-gonad-mesonephros (AGM) region and subsequently relocate to fetal liver. Runx1 transcription factor is essential for HSC development, but is largely dispensable for adult HSCs. Here, we studied tamoxifen-inducible Runx1 inactivation in vivo. Induction at pre-liver stages (up to embryonic day 10.5) reduced erythromyeloid progenitor numbers, but surprisingly did not block the appearance of Runx1-null HSCs in liver. By contrast, ex vivo analysis showed an absolute Runx1 dependency of HSC development in the AGM region. We found that, contrary to current beliefs, significant Cre-inducing tamoxifen activity persists in mouse blood for at least 72 hr after injection. This deferred recombination can hit healthy HSCs, which escaped early Runx1 ablation and result in appearance of Runx1-null HSCs in liver. Such extended recombination activity in vivo is a potential source of misinterpretation, particularly in analysis of dynamic developmental processes during embryogenesis.

Original language	English
Pages (from-to)	784-794
Number of pages	11
Journal	Stem Cell Reports
Volume	11
Issue number	3
DOIs	https://doi.org/10.1016/j.stemcr.2018.08.004
Publication status	Published - 11 Sept 2018

Keywords / Materials (for Non-textual outputs)

AGM
Runx1
conditional knockout
development
hematopoiesis
hematopoietic stem cells
tamoxifen

Access to Document

10.1016/j.stemcr.2018.08.004

1-s2.0-S2213671118303254-main
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
Final published version, 1.72 MBLicence: CC BY-NC-ND

Tracking the embryonic origin of the adult haematopoietic system
Balic, A. & Sang, H.
MRC
1/09/14 → 30/11/18
Project: Research

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@article{39f9c37afe1f47dfb994c633a2e1e566,

title = "Analysis of Runx1 Using Induced Gene Ablation Reveals Its Essential Role in Pre-liver HSC Development and Limitations of an In Vivo Approach",

abstract = "Hematopoietic stem cells (HSCs) develop in the embryonic aorta-gonad-mesonephros (AGM) region and subsequently relocate to fetal liver. Runx1 transcription factor is essential for HSC development, but is largely dispensable for adult HSCs. Here, we studied tamoxifen-inducible Runx1 inactivation in vivo. Induction at pre-liver stages (up to embryonic day 10.5) reduced erythromyeloid progenitor numbers, but surprisingly did not block the appearance of Runx1-null HSCs in liver. By contrast, ex vivo analysis showed an absolute Runx1 dependency of HSC development in the AGM region. We found that, contrary to current beliefs, significant Cre-inducing tamoxifen activity persists in mouse blood for at least 72 hr after injection. This deferred recombination can hit healthy HSCs, which escaped early Runx1 ablation and result in appearance of Runx1-null HSCs in liver. Such extended recombination activity in vivo is a potential source of misinterpretation, particularly in analysis of dynamic developmental processes during embryogenesis.",

keywords = "AGM, Runx1, conditional knockout, development, hematopoiesis, hematopoietic stem cells, tamoxifen",

author = "Jordi Senserrich and Antoniana Batsivari and Stanislav Rybtsov and Sabrina Gordon-Keylock and Celine Souilhol and Frank Buchholz and David Hills and Suling Zhao and Alexander Medvinsky",

note = "Copyright {\textcopyright} 2018. Published by Elsevier Inc.",

year = "2018",

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doi = "10.1016/j.stemcr.2018.08.004",

language = "English",

volume = "11",

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T1 - Analysis of Runx1 Using Induced Gene Ablation Reveals Its Essential Role in Pre-liver HSC Development and Limitations of an In Vivo Approach

AU - Senserrich, Jordi

AU - Batsivari, Antoniana

AU - Rybtsov, Stanislav

AU - Gordon-Keylock, Sabrina

AU - Souilhol, Celine

AU - Buchholz, Frank

AU - Hills, David

AU - Zhao, Suling

AU - Medvinsky, Alexander

PY - 2018/9/11

Y1 - 2018/9/11

N2 - Hematopoietic stem cells (HSCs) develop in the embryonic aorta-gonad-mesonephros (AGM) region and subsequently relocate to fetal liver. Runx1 transcription factor is essential for HSC development, but is largely dispensable for adult HSCs. Here, we studied tamoxifen-inducible Runx1 inactivation in vivo. Induction at pre-liver stages (up to embryonic day 10.5) reduced erythromyeloid progenitor numbers, but surprisingly did not block the appearance of Runx1-null HSCs in liver. By contrast, ex vivo analysis showed an absolute Runx1 dependency of HSC development in the AGM region. We found that, contrary to current beliefs, significant Cre-inducing tamoxifen activity persists in mouse blood for at least 72 hr after injection. This deferred recombination can hit healthy HSCs, which escaped early Runx1 ablation and result in appearance of Runx1-null HSCs in liver. Such extended recombination activity in vivo is a potential source of misinterpretation, particularly in analysis of dynamic developmental processes during embryogenesis.

AB - Hematopoietic stem cells (HSCs) develop in the embryonic aorta-gonad-mesonephros (AGM) region and subsequently relocate to fetal liver. Runx1 transcription factor is essential for HSC development, but is largely dispensable for adult HSCs. Here, we studied tamoxifen-inducible Runx1 inactivation in vivo. Induction at pre-liver stages (up to embryonic day 10.5) reduced erythromyeloid progenitor numbers, but surprisingly did not block the appearance of Runx1-null HSCs in liver. By contrast, ex vivo analysis showed an absolute Runx1 dependency of HSC development in the AGM region. We found that, contrary to current beliefs, significant Cre-inducing tamoxifen activity persists in mouse blood for at least 72 hr after injection. This deferred recombination can hit healthy HSCs, which escaped early Runx1 ablation and result in appearance of Runx1-null HSCs in liver. Such extended recombination activity in vivo is a potential source of misinterpretation, particularly in analysis of dynamic developmental processes during embryogenesis.

KW - AGM

KW - Runx1

KW - conditional knockout

KW - development

KW - hematopoiesis

KW - hematopoietic stem cells

KW - tamoxifen

U2 - 10.1016/j.stemcr.2018.08.004

DO - 10.1016/j.stemcr.2018.08.004

M3 - Article

C2 - 30208304

SN - 2213-6711

VL - 11

SP - 784

EP - 794

JO - Stem Cell Reports

JF - Stem Cell Reports

IS - 3

ER -

Analysis of Runx1 Using Induced Gene Ablation Reveals Its Essential Role in Pre-liver HSC Development and Limitations of an In Vivo Approach

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Keywords / Materials (for Non-textual outputs)

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Tracking the embryonic origin of the adult haematopoietic system

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