High ploidy large cytoplasmic megakaryocytes are hematopoietic stem cells regulators and essential for platelet production

Shen Y. Heazlewood; Tanveer Ahmad; Benjamin Cao; Huimin Cao; Melanie Domingues; Xuan Sun; Chad K. Heazlewood; Songhui Li; Brenda Williams; Madeline Fulton; Jacinta F. White; Tom Nebl; Christian M. Nefzger; Jose M. Polo; Benjamin T. Kile; Felix Kraus; Michael T. Ryan; Yu B. Sun; Peter F.M. Choong; Sarah L. Ellis; Minna Liisa Anko; Susan K. Nilsson

doi:10.1038/s41467-023-37780-7

High ploidy large cytoplasmic megakaryocytes are hematopoietic stem cells regulators and essential for platelet production

Shen Y. Heazlewood
, Tanveer Ahmad
, Benjamin Cao
, Huimin Cao
, Melanie Domingues
, Xuan Sun
, Chad K. Heazlewood
, Songhui Li
, Brenda Williams
, Madeline Fulton
, Jacinta F. White
, Tom Nebl
, Christian M. Nefzger
, Jose M. Polo
, Benjamin T. Kile
, Felix Kraus
, Michael T. Ryan
, Yu B. Sun
, Peter F.M. Choong
, Sarah L. Ellis

Minna Liisa Anko, Susan K. Nilsson^*

^*Corresponding author for this work

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

38 Citations (Scopus)

26 Downloads (Pure)

Abstract

Megakaryocytes (MK) generate platelets. Recently, we and others, have reported MK also regulate hematopoietic stem cells (HSC). Here we show high ploidy large cytoplasmic megakaryocytes (LCM) are critical negative regulators of HSC and critical for platelet formation. Using a mouse knockout model (Pf4-Srsf3Δ/Δ) with normal MK numbers, but essentially devoid of LCM, we demonstrate a pronounced increase in BM HSC concurrent with endogenous mobilization and extramedullary hematopoiesis. Severe thrombocytopenia is observed in animals with diminished LCM, although there is no change in MK ploidy distribution, uncoupling endoreduplication and platelet production. When HSC isolated from a microenvironment essentially devoid of LCM reconstitute hematopoiesis in lethally irradiated mice, the absence of LCM increases HSC in BM, blood and spleen, and the recapitulation of thrombocytopenia. In contrast, following a competitive transplant using minimal numbers of WT HSC together with HSC from a microenvironment with diminished LCM, sufficient WT HSC-generated LCM regulates a normal HSC pool and prevents thrombocytopenia. Importantly, LCM are conserved in humans.

Original language	English
Article number	2099
Number of pages	18
Journal	Nature Communications
Volume	14
Issue number	1
DOIs	https://doi.org/10.1038/s41467-023-37780-7
Publication status	Published - 13 Apr 2023
Publication type	A1 Journal article-refereed

Funding

The authors acknowledge technical assistance from Jessica Hatwell-Humble, Daniela Cardozo and Andrea Reitsma as well as staff and use of facilities at FlowCore, Monash Micro Imaging and Monash Animal Research Platform (Monash University, Australia). The authors thank Patrick Tam for RFP mice, Klaus Matthaei for GFP mice, Angela Cochrane and Michelle Dowsey for co-ordinating the provision and collection of human samples and Samir Taoudi for his rigorous discussion. We acknowledge Ana Oteiza for her contribution in generating some of the schematics. The Australian Regenerative Medicine Institute is supported by grants from the State Government of Victoria and the Australian Government. SKN is supported by a CSIRO OCE Science Leader Fellowship. The authors acknowledge technical assistance from Jessica Hatwell-Humble, Daniela Cardozo and Andrea Reitsma as well as staff and use of facilities at FlowCore, Monash Micro Imaging and Monash Animal Research Platform (Monash University, Australia). The authors thank Patrick Tam for RFP mice, Klaus Matthaei for GFP mice, Angela Cochrane and Michelle Dowsey for co-ordinating the provision and collection of human samples and Samir Taoudi for his rigorous discussion. We acknowledge Ana Oteiza for her contribution in generating some of the schematics. The Australian Regenerative Medicine Institute is supported by grants from the State Government of Victoria and the Australian Government. SKN is supported by a CSIRO OCE Science Leader Fellowship.

Publication forum classification

Publication forum level 3

ASJC Scopus subject areas

General Chemistry
General Biochemistry,Genetics and Molecular Biology
General
General Physics and Astronomy

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10.1038/s41467-023-37780-7Licence: CC BY

s41467-023-37780-7-1Final published version, 4.5 MBLicence: CC BY

https://urn.fi/URN:NBN:fi:tuni-202305316351Licence: CC BY

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Heazlewood, S. Y., Ahmad, T., Cao, B., Cao, H., Domingues, M., Sun, X., Heazlewood, C. K., Li, S., Williams, B., Fulton, M., White, J. F., Nebl, T., Nefzger, C. M., Polo, J. M., Kile, B. T., Kraus, F., Ryan, M. T., Sun, Y. B., Choong, P. F. M., ... Nilsson, S. K. (2023). High ploidy large cytoplasmic megakaryocytes are hematopoietic stem cells regulators and essential for platelet production. Nature Communications, 14(1), Article 2099. https://doi.org/10.1038/s41467-023-37780-7

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title = "High ploidy large cytoplasmic megakaryocytes are hematopoietic stem cells regulators and essential for platelet production",

abstract = "Megakaryocytes (MK) generate platelets. Recently, we and others, have reported MK also regulate hematopoietic stem cells (HSC). Here we show high ploidy large cytoplasmic megakaryocytes (LCM) are critical negative regulators of HSC and critical for platelet formation. Using a mouse knockout model (Pf4-Srsf3Δ/Δ) with normal MK numbers, but essentially devoid of LCM, we demonstrate a pronounced increase in BM HSC concurrent with endogenous mobilization and extramedullary hematopoiesis. Severe thrombocytopenia is observed in animals with diminished LCM, although there is no change in MK ploidy distribution, uncoupling endoreduplication and platelet production. When HSC isolated from a microenvironment essentially devoid of LCM reconstitute hematopoiesis in lethally irradiated mice, the absence of LCM increases HSC in BM, blood and spleen, and the recapitulation of thrombocytopenia. In contrast, following a competitive transplant using minimal numbers of WT HSC together with HSC from a microenvironment with diminished LCM, sufficient WT HSC-generated LCM regulates a normal HSC pool and prevents thrombocytopenia. Importantly, LCM are conserved in humans. ",

author = "Heazlewood, \{Shen Y.\} and Tanveer Ahmad and Benjamin Cao and Huimin Cao and Melanie Domingues and Xuan Sun and Heazlewood, \{Chad K.\} and Songhui Li and Brenda Williams and Madeline Fulton and White, \{Jacinta F.\} and Tom Nebl and Nefzger, \{Christian M.\} and Polo, \{Jose M.\} and Kile, \{Benjamin T.\} and Felix Kraus and Ryan, \{Michael T.\} and Sun, \{Yu B.\} and Choong, \{Peter F.M.\} and Ellis, \{Sarah L.\} and Anko, \{Minna Liisa\} and Nilsson, \{Susan K.\}",

note = "Funding Information: The authors acknowledge technical assistance from Jessica Hatwell-Humble, Daniela Cardozo and Andrea Reitsma as well as staff and use of facilities at FlowCore, Monash Micro Imaging and Monash Animal Research Platform (Monash University, Australia). The authors thank Patrick Tam for RFP mice, Klaus Matthaei for GFP mice, Angela Cochrane and Michelle Dowsey for co-ordinating the provision and collection of human samples and Samir Taoudi for his rigorous discussion. We acknowledge Ana Oteiza for her contribution in generating some of the schematics. The Australian Regenerative Medicine Institute is supported by grants from the State Government of Victoria and the Australian Government. SKN is supported by a CSIRO OCE Science Leader Fellowship. Funding Information: The authors acknowledge technical assistance from Jessica Hatwell-Humble, Daniela Cardozo and Andrea Reitsma as well as staff and use of facilities at FlowCore, Monash Micro Imaging and Monash Animal Research Platform (Monash University, Australia). The authors thank Patrick Tam for RFP mice, Klaus Matthaei for GFP mice, Angela Cochrane and Michelle Dowsey for co-ordinating the provision and collection of human samples and Samir Taoudi for his rigorous discussion. We acknowledge Ana Oteiza for her contribution in generating some of the schematics. The Australian Regenerative Medicine Institute is supported by grants from the State Government of Victoria and the Australian Government. SKN is supported by a CSIRO OCE Science Leader Fellowship. Publisher Copyright: {\textcopyright} 2023, Crown.",

year = "2023",

month = apr,

day = "13",

doi = "10.1038/s41467-023-37780-7",

language = "English",

volume = "14",

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issn = "2041-1723",

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Heazlewood, SY, Ahmad, T, Cao, B, Cao, H, Domingues, M, Sun, X, Heazlewood, CK, Li, S, Williams, B, Fulton, M, White, JF, Nebl, T, Nefzger, CM, Polo, JM, Kile, BT, Kraus, F, Ryan, MT, Sun, YB, Choong, PFM, Ellis, SL, Anko, ML & Nilsson, SK 2023, 'High ploidy large cytoplasmic megakaryocytes are hematopoietic stem cells regulators and essential for platelet production', Nature Communications, vol. 14, no. 1, 2099. https://doi.org/10.1038/s41467-023-37780-7

TY - JOUR

T1 - High ploidy large cytoplasmic megakaryocytes are hematopoietic stem cells regulators and essential for platelet production

AU - Heazlewood, Shen Y.

AU - Ahmad, Tanveer

AU - Cao, Benjamin

AU - Cao, Huimin

AU - Domingues, Melanie

AU - Sun, Xuan

AU - Heazlewood, Chad K.

AU - Li, Songhui

AU - Williams, Brenda

AU - Fulton, Madeline

AU - White, Jacinta F.

AU - Nebl, Tom

AU - Nefzger, Christian M.

AU - Polo, Jose M.

AU - Kile, Benjamin T.

AU - Kraus, Felix

AU - Ryan, Michael T.

AU - Sun, Yu B.

AU - Choong, Peter F.M.

AU - Ellis, Sarah L.

AU - Anko, Minna Liisa

AU - Nilsson, Susan K.

N1 - Funding Information: The authors acknowledge technical assistance from Jessica Hatwell-Humble, Daniela Cardozo and Andrea Reitsma as well as staff and use of facilities at FlowCore, Monash Micro Imaging and Monash Animal Research Platform (Monash University, Australia). The authors thank Patrick Tam for RFP mice, Klaus Matthaei for GFP mice, Angela Cochrane and Michelle Dowsey for co-ordinating the provision and collection of human samples and Samir Taoudi for his rigorous discussion. We acknowledge Ana Oteiza for her contribution in generating some of the schematics. The Australian Regenerative Medicine Institute is supported by grants from the State Government of Victoria and the Australian Government. SKN is supported by a CSIRO OCE Science Leader Fellowship. Funding Information: The authors acknowledge technical assistance from Jessica Hatwell-Humble, Daniela Cardozo and Andrea Reitsma as well as staff and use of facilities at FlowCore, Monash Micro Imaging and Monash Animal Research Platform (Monash University, Australia). The authors thank Patrick Tam for RFP mice, Klaus Matthaei for GFP mice, Angela Cochrane and Michelle Dowsey for co-ordinating the provision and collection of human samples and Samir Taoudi for his rigorous discussion. We acknowledge Ana Oteiza for her contribution in generating some of the schematics. The Australian Regenerative Medicine Institute is supported by grants from the State Government of Victoria and the Australian Government. SKN is supported by a CSIRO OCE Science Leader Fellowship. Publisher Copyright: © 2023, Crown.

PY - 2023/4/13

Y1 - 2023/4/13

N2 - Megakaryocytes (MK) generate platelets. Recently, we and others, have reported MK also regulate hematopoietic stem cells (HSC). Here we show high ploidy large cytoplasmic megakaryocytes (LCM) are critical negative regulators of HSC and critical for platelet formation. Using a mouse knockout model (Pf4-Srsf3Δ/Δ) with normal MK numbers, but essentially devoid of LCM, we demonstrate a pronounced increase in BM HSC concurrent with endogenous mobilization and extramedullary hematopoiesis. Severe thrombocytopenia is observed in animals with diminished LCM, although there is no change in MK ploidy distribution, uncoupling endoreduplication and platelet production. When HSC isolated from a microenvironment essentially devoid of LCM reconstitute hematopoiesis in lethally irradiated mice, the absence of LCM increases HSC in BM, blood and spleen, and the recapitulation of thrombocytopenia. In contrast, following a competitive transplant using minimal numbers of WT HSC together with HSC from a microenvironment with diminished LCM, sufficient WT HSC-generated LCM regulates a normal HSC pool and prevents thrombocytopenia. Importantly, LCM are conserved in humans.

AB - Megakaryocytes (MK) generate platelets. Recently, we and others, have reported MK also regulate hematopoietic stem cells (HSC). Here we show high ploidy large cytoplasmic megakaryocytes (LCM) are critical negative regulators of HSC and critical for platelet formation. Using a mouse knockout model (Pf4-Srsf3Δ/Δ) with normal MK numbers, but essentially devoid of LCM, we demonstrate a pronounced increase in BM HSC concurrent with endogenous mobilization and extramedullary hematopoiesis. Severe thrombocytopenia is observed in animals with diminished LCM, although there is no change in MK ploidy distribution, uncoupling endoreduplication and platelet production. When HSC isolated from a microenvironment essentially devoid of LCM reconstitute hematopoiesis in lethally irradiated mice, the absence of LCM increases HSC in BM, blood and spleen, and the recapitulation of thrombocytopenia. In contrast, following a competitive transplant using minimal numbers of WT HSC together with HSC from a microenvironment with diminished LCM, sufficient WT HSC-generated LCM regulates a normal HSC pool and prevents thrombocytopenia. Importantly, LCM are conserved in humans.

U2 - 10.1038/s41467-023-37780-7

DO - 10.1038/s41467-023-37780-7

M3 - Article

C2 - 37055407

AN - SCOPUS:85152471293

SN - 2041-1723

VL - 14

JO - Nature Communications

JF - Nature Communications

IS - 1

M1 - 2099

ER -

High ploidy large cytoplasmic megakaryocytes are hematopoietic stem cells regulators and essential for platelet production

Abstract

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