Bioengineered niches that recreate physiological extracellular matrix organisation to support long-term haematopoietic stem cells

Hannah Donnelly; Ewan Ross; Yinbo Xiao; Rio Hermantara; Aqeel F Taqi; W Sebastian Doherty-Boyd; Jennifer Cassels; Penelope M Tsimbouri; Karen M Dunn; Jodie Hay; Annie Cheng; R M Dominic Meek; Nikhil Jain; Christopher West; Helen Wheadon; Alison M Michie; Bruno Peault; Adam G West; Manuel Salmeron-Sanchez; Matthew J Dalby

doi:10.1038/s41467-024-50054-0

Bioengineered niches that recreate physiological extracellular matrix organisation to support long-term haematopoietic stem cells

Hannah Donnelly, Ewan Ross, Yinbo Xiao, Rio Hermantara, Aqeel F Taqi, W Sebastian Doherty-Boyd, Jennifer Cassels, Penelope M Tsimbouri, Karen M Dunn, Jodie Hay, Annie Cheng, R M Dominic Meek, Nikhil Jain, Christopher West, Helen Wheadon, Alison M Michie, Bruno Peault, Adam G West, Manuel Salmeron-Sanchez, Matthew J Dalby

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

Abstract

Long-term reconstituting haematopoietic stem cells (LT-HSCs) are used to treat blood disorders via stem cell transplantation. The very low abundance of LT-HSCs and their rapid differentiation during in vitro culture hinders their clinical utility. Previous developments using stromal feeder layers, defined media cocktails, and bioengineering have enabled HSC expansion in culture, but of mostly short-term HSCs and progenitor populations at the expense of naive LT-HSCs. Here, we report the creation of a bioengineered LT-HSC maintenance niche that recreates physiological extracellular matrix organisation, using soft collagen type-I hydrogels to drive nestin expression in perivascular stromal cells (PerSCs). We demonstrate that nestin, which is expressed by HSC-supportive bone marrow stromal cells, is cytoprotective and, via regulation of metabolism, is important for HIF-1α expression in PerSCs. When CD34+ve HSCs were added to the bioengineered niches comprising nestin/HIF-1α expressing PerSCs, LT-HSC numbers were maintained with normal clonal and in vivo reconstitution potential, without media supplementation. We provide proof-of-concept that our bioengineered niches can support the survival of CRISPR edited HSCs. Successful editing of LT-HSCs ex vivo can have potential impact on the treatment of blood disorders.

Original language	English
Article number	5791
Number of pages	18
Journal	Nature Communications
Volume	15
DOIs	https://doi.org/10.1038/s41467-024-50054-0
Publication status	Published - 10 Jul 2024

Bibliographical note

Copyright © The Author(s) 2024. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit https://creativecommons.org/licenses/by/4.0/.

Data Access Statement

The LC-MS data and all datasets supporting the findings in this study are openly available under the Creative Commons Attribution (CC-BY) license at: https://doi.org/10.5525/gla.researchdata.1326. Due to the large size of the data files, access must be requested. Access can be obtained from the corresponding authors. Requests will be processed within 2 working days. The RNAseq data generated from PerSCs in this study have been deposited in NCBI’s Gene Expression Omnibus and are accessible through GEO Series accession number GSE265789. Source data are provided with this paper.

The custom CellProfiler pipeline used for nuclear HIF1α measurements is openly available in the DOI file at: https://doi.org/10.5525/gla.researchdata.1326. The data are licenced with a CC-BY open access licence and access requests to the corresponding authors are handled within two working days.

Keywords

Hematopoietic Stem Cells/metabolism
Animals
Nestin/metabolism
Extracellular Matrix/metabolism
Hypoxia-Inducible Factor 1, alpha Subunit/metabolism
Mice
Stem Cell Niche
Hydrogels/chemistry
Bioengineering/methods
Humans
Mesenchymal Stem Cells/metabolism
Hematopoietic Stem Cell Transplantation
Antigens, CD34/metabolism
Collagen Type I/metabolism
Cell Differentiation
Mice, Inbred C57BL

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10.1038/s41467-024-50054-0Licence: CC BY 4.0

Bioengineered niches that recreate physiological extracellular matrix organisation to support long-term haematopoietic stem cells
Copyright © The Author(s) 2024. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit https://creativecommons.org/licenses/by/4.0/.
Final published version, 7.1 MBLicence: CC BY 4.0

Cite this

Donnelly, H., Ross, E., Xiao, Y., Hermantara, R., Taqi, A. F., Doherty-Boyd, W. S., Cassels, J., Tsimbouri, P. M., Dunn, K. M., Hay, J., Cheng, A., Meek, R. M. D., Jain, N., West, C., Wheadon, H., Michie, A. M., Peault, B., West, A. G., Salmeron-Sanchez, M., & Dalby, M. J. (2024). Bioengineered niches that recreate physiological extracellular matrix organisation to support long-term haematopoietic stem cells. Nature Communications, 15, Article 5791. https://doi.org/10.1038/s41467-024-50054-0

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abstract = "Long-term reconstituting haematopoietic stem cells (LT-HSCs) are used to treat blood disorders via stem cell transplantation. The very low abundance of LT-HSCs and their rapid differentiation during in vitro culture hinders their clinical utility. Previous developments using stromal feeder layers, defined media cocktails, and bioengineering have enabled HSC expansion in culture, but of mostly short-term HSCs and progenitor populations at the expense of naive LT-HSCs. Here, we report the creation of a bioengineered LT-HSC maintenance niche that recreates physiological extracellular matrix organisation, using soft collagen type-I hydrogels to drive nestin expression in perivascular stromal cells (PerSCs). We demonstrate that nestin, which is expressed by HSC-supportive bone marrow stromal cells, is cytoprotective and, via regulation of metabolism, is important for HIF-1α expression in PerSCs. When CD34+ve HSCs were added to the bioengineered niches comprising nestin/HIF-1α expressing PerSCs, LT-HSC numbers were maintained with normal clonal and in vivo reconstitution potential, without media supplementation. We provide proof-of-concept that our bioengineered niches can support the survival of CRISPR edited HSCs. Successful editing of LT-HSCs ex vivo can have potential impact on the treatment of blood disorders.",

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author = "Hannah Donnelly and Ewan Ross and Yinbo Xiao and Rio Hermantara and Taqi, {Aqeel F} and Doherty-Boyd, {W Sebastian} and Jennifer Cassels and Tsimbouri, {Penelope M} and Dunn, {Karen M} and Jodie Hay and Annie Cheng and Meek, {R M Dominic} and Nikhil Jain and Christopher West and Helen Wheadon and Michie, {Alison M} and Bruno Peault and West, {Adam G} and Manuel Salmeron-Sanchez and Dalby, {Matthew J}",

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Donnelly, H, Ross, E, Xiao, Y, Hermantara, R, Taqi, AF, Doherty-Boyd, WS, Cassels, J, Tsimbouri, PM, Dunn, KM, Hay, J, Cheng, A, Meek, RMD, Jain, N, West, C, Wheadon, H, Michie, AM, Peault, B, West, AG, Salmeron-Sanchez, M & Dalby, MJ 2024, 'Bioengineered niches that recreate physiological extracellular matrix organisation to support long-term haematopoietic stem cells', Nature Communications, vol. 15, 5791. https://doi.org/10.1038/s41467-024-50054-0

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AU - Donnelly, Hannah

AU - Ross, Ewan

AU - Xiao, Yinbo

AU - Hermantara, Rio

AU - Taqi, Aqeel F

AU - Doherty-Boyd, W Sebastian

AU - Cassels, Jennifer

AU - Tsimbouri, Penelope M

AU - Dunn, Karen M

AU - Hay, Jodie

AU - Cheng, Annie

AU - Meek, R M Dominic

AU - Jain, Nikhil

AU - West, Christopher

AU - Wheadon, Helen

AU - Michie, Alison M

AU - Peault, Bruno

AU - West, Adam G

AU - Salmeron-Sanchez, Manuel

AU - Dalby, Matthew J

N1 - Copyright © The Author(s) 2024. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit https://creativecommons.org/licenses/by/4.0/.

PY - 2024/7/10

Y1 - 2024/7/10

N2 - Long-term reconstituting haematopoietic stem cells (LT-HSCs) are used to treat blood disorders via stem cell transplantation. The very low abundance of LT-HSCs and their rapid differentiation during in vitro culture hinders their clinical utility. Previous developments using stromal feeder layers, defined media cocktails, and bioengineering have enabled HSC expansion in culture, but of mostly short-term HSCs and progenitor populations at the expense of naive LT-HSCs. Here, we report the creation of a bioengineered LT-HSC maintenance niche that recreates physiological extracellular matrix organisation, using soft collagen type-I hydrogels to drive nestin expression in perivascular stromal cells (PerSCs). We demonstrate that nestin, which is expressed by HSC-supportive bone marrow stromal cells, is cytoprotective and, via regulation of metabolism, is important for HIF-1α expression in PerSCs. When CD34+ve HSCs were added to the bioengineered niches comprising nestin/HIF-1α expressing PerSCs, LT-HSC numbers were maintained with normal clonal and in vivo reconstitution potential, without media supplementation. We provide proof-of-concept that our bioengineered niches can support the survival of CRISPR edited HSCs. Successful editing of LT-HSCs ex vivo can have potential impact on the treatment of blood disorders.

AB - Long-term reconstituting haematopoietic stem cells (LT-HSCs) are used to treat blood disorders via stem cell transplantation. The very low abundance of LT-HSCs and their rapid differentiation during in vitro culture hinders their clinical utility. Previous developments using stromal feeder layers, defined media cocktails, and bioengineering have enabled HSC expansion in culture, but of mostly short-term HSCs and progenitor populations at the expense of naive LT-HSCs. Here, we report the creation of a bioengineered LT-HSC maintenance niche that recreates physiological extracellular matrix organisation, using soft collagen type-I hydrogels to drive nestin expression in perivascular stromal cells (PerSCs). We demonstrate that nestin, which is expressed by HSC-supportive bone marrow stromal cells, is cytoprotective and, via regulation of metabolism, is important for HIF-1α expression in PerSCs. When CD34+ve HSCs were added to the bioengineered niches comprising nestin/HIF-1α expressing PerSCs, LT-HSC numbers were maintained with normal clonal and in vivo reconstitution potential, without media supplementation. We provide proof-of-concept that our bioengineered niches can support the survival of CRISPR edited HSCs. Successful editing of LT-HSCs ex vivo can have potential impact on the treatment of blood disorders.

KW - Hematopoietic Stem Cells/metabolism

KW - Animals

KW - Nestin/metabolism

KW - Extracellular Matrix/metabolism

KW - Hypoxia-Inducible Factor 1, alpha Subunit/metabolism

KW - Mice

KW - Stem Cell Niche

KW - Hydrogels/chemistry

KW - Bioengineering/methods

KW - Humans

KW - Mesenchymal Stem Cells/metabolism

KW - Hematopoietic Stem Cell Transplantation

KW - Antigens, CD34/metabolism

KW - Collagen Type I/metabolism

KW - Cell Differentiation

KW - Mice, Inbred C57BL

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DO - 10.1038/s41467-024-50054-0

M3 - Article

C2 - 38987295

SN - 2041-1723

VL - 15

JO - Nature Communications

JF - Nature Communications

M1 - 5791

ER -

Bioengineered niches that recreate physiological extracellular matrix organisation to support long-term haematopoietic stem cells

Abstract

Bibliographical note

Data Access Statement

Keywords

Access to Document

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