RSPO3 Furin domain-conjugated liposomes for selective drug delivery to LGR5-high cells

Peter van Kerkhof; Tomica Kralj; Francesca Spanevello; Louis van Bloois; Ingrid Jordens; Jelte van der Vaart; Cara Jamieson; Alessandra Merenda; Enrico Mastrobattista; Madelon M Maurice

doi:10.1016/j.jconrel.2023.02.025

RSPO3 Furin domain-conjugated liposomes for selective drug delivery to LGR5-high cells

Peter van Kerkhof, Tomica Kralj, Francesca Spanevello, Louis van Bloois, Ingrid Jordens, Jelte van der Vaart, Cara Jamieson, Alessandra Merenda, Enrico Mastrobattista^*, Madelon M Maurice^*

^*Corresponding author for this work

University Medical Center Utrecht

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

Abstract

The transmembrane receptor LGR5 potentiates Wnt/β-catenin signaling by binding both secreted R-spondin (RSPOs) and the Wnt tumor suppressors RNF43/ZNRF3, directing clearance of RNF43/ZNRF3 from the cell surface. Besides being widely used as a stem cell marker in various tissues, LGR5 is overexpressed in many types of malignancies, including colorectal cancer. Its expression characterizes a subpopulation of cancer cells that play a crucial role in tumor initiation, progression and cancer relapse, known as cancer stem cells (CSCs). For this reason, ongoing efforts are aimed at eradicating LGR5-positive CSCs. Here, we engineered liposomes decorated with different RSPO proteins to specifically detect and target LGR5-positive cells. Using fluorescence-loaded liposomes, we show that conjugation of full-length RSPO1 to the liposomal surface mediates aspecific, LGR5-independent cellular uptake, largely mediated by heparan sulfate proteoglycan binding. By contrast, liposomes decorated only with the Furin (FuFu) domains of RSPO3 are taken up by cells in a highly specific, LGR5-dependent manner. Moreover, encapsulating doxorubicin in FuFuRSPO3 liposomes allowed us to selectively inhibit the growth of LGR5-high cells. Thus, FuFuRSPO3-coated liposomes allow for the selective detection and ablation of LGR5-high cells, providing a potential drug delivery system for LGR5-targeted anti-cancer strategies.

Original language	English
Pages (from-to)	72-83
Number of pages	12
Journal	Journal of controlled release : official journal of the Controlled Release Society
Volume	356
DOIs	https://doi.org/10.1016/j.jconrel.2023.02.025
Publication status	Published - Apr 2023

Bibliographical note

Publisher Copyright:
© 2023 The Authors

Funding

We thank members of the laboratory of M.M.M. for discussions and suggestions, Dieuwke Marvin for technical assistance and Huiying Ma for the generation of the mouse small intestinal APCmut organoids. Rajat Rohatgi for the WT and LGR4/5/6 triple knockout HAP1 cells (Beckman center, Stanford). Bruno Reversade for the WT and LGR4/5/6 triple knockout MEF cells (A*STAR, Singapore). Niels de Wind for the LoVo cells (LUMC, Leiden) and Bart Spee for the Huh7 cells (UU, Utrecht). This work is part of the Oncode Institute, which is partly financed by the Dutch Cancer Society. This work was supported by the Netherlands Organization for Scientific Research NWO VICI Grant 91815604, TOP Grant 91218050 (to M.M.M.) and a Seed grant from Utrecht University (to M.M.M. and E.M.). We thank members of the laboratory of M.M.M. for discussions and suggestions, Dieuwke Marvin for technical assistance and Huiying Ma for the generation of the mouse small intestinal APC mut organoids. Rajat Rohatgi for the WT and LGR4/5/6 triple knockout HAP1 cells (Beckman center, Stanford). Bruno Reversade for the WT and LGR4/5/6 triple knockout MEF cells (A*STAR, Singapore). Niels de Wind for the LoVo cells (LUMC, Leiden) and Bart Spee for the Huh7 cells (UU, Utrecht). This work is part of the Oncode Institute, which is partly financed by the Dutch Cancer Society. This work was supported by the Netherlands Organization for Scientific Research NWO VICI Grant 91815604 , TOP Grant 91218050 (to M.M.M.) and a Seed grant from Utrecht University (to M.M.M. and E.M.).

Funders	Funder number
Bart Spee for the Huh7 cells
Technology Opportunities Program	91218050
Agency for Science, Technology and Research
Universiteit Utrecht
Nederlandse Organisatie voor Wetenschappelijk Onderzoek	91815604
KWF Kankerbestrijding
Leids Universitair Medisch Centrum
Oncode Institute

Keywords

Cancer stem cells
Doxorubicin
Drug delivery
LGR5
Liposomes
R-spondin-3

UN SDGs

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

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10.1016/j.jconrel.2023.02.025Licence: CC BY

1-s2.0-S0168365923001360-mainFinal published version, 3.25 MBLicence: CC BY

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van Kerkhof, P., Kralj, T., Spanevello, F., van Bloois, L., Jordens, I., van der Vaart, J., Jamieson, C., Merenda, A., Mastrobattista, E., & Maurice, M. M. (2023). RSPO3 Furin domain-conjugated liposomes for selective drug delivery to LGR5-high cells. Journal of controlled release : official journal of the Controlled Release Society, 356, 72-83. https://doi.org/10.1016/j.jconrel.2023.02.025

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title = "RSPO3 Furin domain-conjugated liposomes for selective drug delivery to LGR5-high cells",

abstract = "The transmembrane receptor LGR5 potentiates Wnt/β-catenin signaling by binding both secreted R-spondin (RSPOs) and the Wnt tumor suppressors RNF43/ZNRF3, directing clearance of RNF43/ZNRF3 from the cell surface. Besides being widely used as a stem cell marker in various tissues, LGR5 is overexpressed in many types of malignancies, including colorectal cancer. Its expression characterizes a subpopulation of cancer cells that play a crucial role in tumor initiation, progression and cancer relapse, known as cancer stem cells (CSCs). For this reason, ongoing efforts are aimed at eradicating LGR5-positive CSCs. Here, we engineered liposomes decorated with different RSPO proteins to specifically detect and target LGR5-positive cells. Using fluorescence-loaded liposomes, we show that conjugation of full-length RSPO1 to the liposomal surface mediates aspecific, LGR5-independent cellular uptake, largely mediated by heparan sulfate proteoglycan binding. By contrast, liposomes decorated only with the Furin (FuFu) domains of RSPO3 are taken up by cells in a highly specific, LGR5-dependent manner. Moreover, encapsulating doxorubicin in FuFuRSPO3 liposomes allowed us to selectively inhibit the growth of LGR5-high cells. Thus, FuFuRSPO3-coated liposomes allow for the selective detection and ablation of LGR5-high cells, providing a potential drug delivery system for LGR5-targeted anti-cancer strategies.",

keywords = "Cancer stem cells, Doxorubicin, Drug delivery, LGR5, Liposomes, R-spondin-3",

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doi = "10.1016/j.jconrel.2023.02.025",

language = "English",

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van Kerkhof, P, Kralj, T, Spanevello, F, van Bloois, L, Jordens, I, van der Vaart, J, Jamieson, C, Merenda, A, Mastrobattista, E & Maurice, MM 2023, 'RSPO3 Furin domain-conjugated liposomes for selective drug delivery to LGR5-high cells', Journal of controlled release : official journal of the Controlled Release Society, vol. 356, pp. 72-83. https://doi.org/10.1016/j.jconrel.2023.02.025

TY - JOUR

T1 - RSPO3 Furin domain-conjugated liposomes for selective drug delivery to LGR5-high cells

AU - van Kerkhof, Peter

AU - Kralj, Tomica

AU - Spanevello, Francesca

AU - van Bloois, Louis

AU - Jordens, Ingrid

AU - van der Vaart, Jelte

AU - Jamieson, Cara

AU - Merenda, Alessandra

AU - Mastrobattista, Enrico

AU - Maurice, Madelon M

PY - 2023/4

Y1 - 2023/4

N2 - The transmembrane receptor LGR5 potentiates Wnt/β-catenin signaling by binding both secreted R-spondin (RSPOs) and the Wnt tumor suppressors RNF43/ZNRF3, directing clearance of RNF43/ZNRF3 from the cell surface. Besides being widely used as a stem cell marker in various tissues, LGR5 is overexpressed in many types of malignancies, including colorectal cancer. Its expression characterizes a subpopulation of cancer cells that play a crucial role in tumor initiation, progression and cancer relapse, known as cancer stem cells (CSCs). For this reason, ongoing efforts are aimed at eradicating LGR5-positive CSCs. Here, we engineered liposomes decorated with different RSPO proteins to specifically detect and target LGR5-positive cells. Using fluorescence-loaded liposomes, we show that conjugation of full-length RSPO1 to the liposomal surface mediates aspecific, LGR5-independent cellular uptake, largely mediated by heparan sulfate proteoglycan binding. By contrast, liposomes decorated only with the Furin (FuFu) domains of RSPO3 are taken up by cells in a highly specific, LGR5-dependent manner. Moreover, encapsulating doxorubicin in FuFuRSPO3 liposomes allowed us to selectively inhibit the growth of LGR5-high cells. Thus, FuFuRSPO3-coated liposomes allow for the selective detection and ablation of LGR5-high cells, providing a potential drug delivery system for LGR5-targeted anti-cancer strategies.

AB - The transmembrane receptor LGR5 potentiates Wnt/β-catenin signaling by binding both secreted R-spondin (RSPOs) and the Wnt tumor suppressors RNF43/ZNRF3, directing clearance of RNF43/ZNRF3 from the cell surface. Besides being widely used as a stem cell marker in various tissues, LGR5 is overexpressed in many types of malignancies, including colorectal cancer. Its expression characterizes a subpopulation of cancer cells that play a crucial role in tumor initiation, progression and cancer relapse, known as cancer stem cells (CSCs). For this reason, ongoing efforts are aimed at eradicating LGR5-positive CSCs. Here, we engineered liposomes decorated with different RSPO proteins to specifically detect and target LGR5-positive cells. Using fluorescence-loaded liposomes, we show that conjugation of full-length RSPO1 to the liposomal surface mediates aspecific, LGR5-independent cellular uptake, largely mediated by heparan sulfate proteoglycan binding. By contrast, liposomes decorated only with the Furin (FuFu) domains of RSPO3 are taken up by cells in a highly specific, LGR5-dependent manner. Moreover, encapsulating doxorubicin in FuFuRSPO3 liposomes allowed us to selectively inhibit the growth of LGR5-high cells. Thus, FuFuRSPO3-coated liposomes allow for the selective detection and ablation of LGR5-high cells, providing a potential drug delivery system for LGR5-targeted anti-cancer strategies.

KW - Cancer stem cells

KW - Doxorubicin

KW - Drug delivery

KW - LGR5

KW - Liposomes

KW - R-spondin-3

UR - http://www.scopus.com/inward/record.url?scp=85149364588&partnerID=8YFLogxK

U2 - 10.1016/j.jconrel.2023.02.025

DO - 10.1016/j.jconrel.2023.02.025

M3 - Article

C2 - 36813038

SN - 0168-3659

VL - 356

SP - 72

EP - 83

JO - Journal of controlled release : official journal of the Controlled Release Society

JF - Journal of controlled release : official journal of the Controlled Release Society

ER -

RSPO3 Furin domain-conjugated liposomes for selective drug delivery to LGR5-high cells

Abstract

Bibliographical note

Funding

Keywords

UN SDGs

Access to Document

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