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Article ; Online: Exciting Times for Lipid Nanoparticles: How Canadian Discoveries Are Enabling Gene Therapies.

Cheng, Miffy H Y / Brimacombe, Cedric A / Verbeke, Rein / Cullis, Pieter R

2022 Volume 19, Issue 6, Page(s) 1663–1668

Abstract: In this brief perspective, we describe key events in the history of the lipid-based nanomedicine field, highlight Canadian contributions, and outline areas where lipid nanoparticle technology is poised to have a transformative effect on the future of ... ...

Abstract	In this brief perspective, we describe key events in the history of the lipid-based nanomedicine field, highlight Canadian contributions, and outline areas where lipid nanoparticle technology is poised to have a transformative effect on the future of medicine.
MeSH term(s)	CRISPR-Cas Systems ; Canada ; Gene Editing ; Liposomes ; Nanoparticles
Chemical Substances	Lipid Nanoparticles ; Liposomes
Language	English
Publishing date	2022-05-18
Publishing country	United States
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ZDB-ID	2138405-8
ISSN	1543-8392 ; 1543-8384
ISSN (online)	1543-8392
ISSN	1543-8384
DOI	10.1021/acs.molpharmaceut.2c00365
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Article ; Online: Innate immune mechanisms of mRNA vaccines.

Verbeke, Rein / Hogan, Michael J / Loré, Karin / Pardi, Norbert

Immunity

2022 Volume 55, Issue 11, Page(s) 1993–2005

Abstract: The lipid nanoparticle (LNP)-encapsulated, nucleoside-modified mRNA platform has been used to generate safe and effective vaccines in record time against COVID-19. Here, we review the current understanding of the manner whereby mRNA vaccines induce ... ...

Abstract	The lipid nanoparticle (LNP)-encapsulated, nucleoside-modified mRNA platform has been used to generate safe and effective vaccines in record time against COVID-19. Here, we review the current understanding of the manner whereby mRNA vaccines induce innate immune activation and how this contributes to protective immunity. We discuss innate immune sensing of mRNA vaccines at the cellular and intracellular levels and consider the contribution of both the mRNA and the LNP components to their immunogenicity. A key message that is emerging from recent observations is that the LNP carrier acts as a powerful adjuvant for this novel vaccine platform. In this context, we highlight important gaps in understanding and discuss how new insight into the mechanisms underlying the effectiveness of mRNA-LNP vaccines may enable tailoring mRNA and carrier molecules to develop vaccines with greater effectiveness and milder adverse events in the future.
MeSH term(s)	Humans ; COVID-19/prevention & control ; Vaccines, Synthetic ; RNA, Messenger/genetics ; Immunity, Innate ; mRNA Vaccines
Chemical Substances	Lipid Nanoparticles ; Vaccines, Synthetic ; RNA, Messenger
Language	English
Publishing date	2022-11-04
Publishing country	United States
Document type	Journal Article ; Review ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	1217235-2
ISSN	1097-4180 ; 1074-7613
ISSN (online)	1097-4180
ISSN	1074-7613
DOI	10.1016/j.immuni.2022.10.014
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Zs.A 4227: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
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Article ; Online: The dawn of mRNA vaccines: The COVID-19 case.

Verbeke, Rein / Lentacker, Ine / De Smedt, Stefaan C / Dewitte, Heleen

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

2021 Volume 333, Page(s) 511–520

Abstract: In less than one year since the outbreak of the COVID-19 pandemic, two mRNA-based vaccines, BNT162b2 and mRNA-1273, were granted the first historic authorization for emergency use, while another mRNA vaccine, CVnCoV, progressed to phase 3 clinical ... ...

Abstract	In less than one year since the outbreak of the COVID-19 pandemic, two mRNA-based vaccines, BNT162b2 and mRNA-1273, were granted the first historic authorization for emergency use, while another mRNA vaccine, CVnCoV, progressed to phase 3 clinical testing. The COVID-19 mRNA vaccines represent a new class of vaccine products, which consist of synthetic mRNA strands encoding the SARS-CoV-2 Spike glycoprotein, packaged in lipid nanoparticles to deliver mRNA to cells. This review digs deeper into the scientific breakthroughs of the last decades that laid the foundations for the rapid rise of mRNA vaccines during the COVID-19 pandemic. As well as providing momentum for mRNA vaccines, SARS-CoV-2 represents an ideal case study allowing to compare design-activity differences between the different mRNA vaccine candidates. Therefore, a detailed overview of the composition and (pre)clinical performance of the three most advanced mRNA vaccines is provided and the influence of choices in their structural design on to their immunogenicity and reactogenicity profile is discussed in depth. In addition to the new fundamental insights in the mRNA vaccines' mode of action highlighted here, we also point out which unknowns remain that require further investigation and possibly, optimization in future mRNA vaccine development.
MeSH term(s)	BNT162 Vaccine ; COVID-19 ; COVID-19 Vaccines ; Humans ; Pandemics ; RNA, Messenger ; SARS-CoV-2 ; Vaccines
Chemical Substances	COVID-19 Vaccines ; RNA, Messenger ; Vaccines ; BNT162 Vaccine (N38TVC63NU)
Language	English
Publishing date	2021-03-30
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	632533-6
ISSN	1873-4995 ; 0168-3659
ISSN (online)	1873-4995
ISSN	0168-3659
DOI	10.1016/j.jconrel.2021.03.043
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Article ; Online: Alpha-galactosylceramide improves the potency of mRNA LNP vaccines against cancer and intracellular bacteria.

Meulewaeter, Sofie / Aernout, Ilke / Deprez, Joke / Engelen, Yanou / De Velder, Margo / Franceschini, Lorenzo / Breckpot, Karine / Van Calenbergh, Serge / Asselman, Caroline / Boucher, Katie / Impens, Francis / De Smedt, Stefaan C / Verbeke, Rein / Lentacker, Ine

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

2024

Abstract: Although various types of mRNA-based vaccines have been explored, the optimal conditions for induction of both humoral and cellular immunity remain rather unknown. In this study, mRNA vaccines of nucleoside-modified mRNA in lipoplexes (LPXs) or lipid ... ...

Abstract	Although various types of mRNA-based vaccines have been explored, the optimal conditions for induction of both humoral and cellular immunity remain rather unknown. In this study, mRNA vaccines of nucleoside-modified mRNA in lipoplexes (LPXs) or lipid nanoparticles (LNPs) were evaluated after administration in mice through different routes, assessing mRNA delivery, tolerability and immunogenicity. In addition, we investigated whether mRNA vaccines could benefit from the inclusion of the adjuvant alpha-galactosylceramide (αGC), an invariant Natural Killer T (NKT) cell ligand. Intramuscular (IM) vaccination with ovalbumin (OVA)-encoding mRNA encapsulated in LNPs adjuvanted with αGC showed the highest antibody- and CD8
Language	English
Publishing date	2024-04-30
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	632533-6
ISSN	1873-4995 ; 0168-3659
ISSN (online)	1873-4995
ISSN	0168-3659
DOI	10.1016/j.jconrel.2024.04.052
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Article ; Online: Continuous freeze-drying of messenger RNA lipid nanoparticles enables storage at higher temperatures.

Meulewaeter, Sofie / Nuytten, Gust / Cheng, Miffy H Y / De Smedt, Stefaan C / Cullis, Pieter R / De Beer, Thomas / Lentacker, Ine / Verbeke, Rein

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

2023 Volume 357, Page(s) 149–160

Abstract: Messenger RNA (mRNA) lipid nanoparticles (LNPs) have emerged at the forefront during the COVID-19 vaccination campaign. Despite their tremendous success, mRNA vaccines currently require storage at deep freeze temperatures which complicates their storage ... ...

Abstract	Messenger RNA (mRNA) lipid nanoparticles (LNPs) have emerged at the forefront during the COVID-19 vaccination campaign. Despite their tremendous success, mRNA vaccines currently require storage at deep freeze temperatures which complicates their storage and distribution, and ultimately leads to lower accessibility to low- and middle-income countries. To elaborate on this challenge, we investigated freeze-drying as a method to enable storage of mRNA LNPs at room- and even higher temperatures. More specifically, we explored a novel continuous freeze-drying technique based on spin-freezing, which has several advantages compared to classical batch freeze-drying including a much shorter drying time and improved process and product quality controlling. Here, we give insight into the variables that play a role during freeze-drying by evaluating the impact of the buffer and mRNA LNP formulation (ionizable lipid to mRNA weight ratio) on properties such as size, morphology and mRNA encapsulation. We found that a sufficiently high ionizable lipid to mRNA weight ratio was necessary to prevent leakage of mRNA during freeze-drying and that phosphate and Tris, but not PBS, were appropriate buffers for lyophilization of mRNA LNPs. We also studied the stability of optimally lyophilized mRNA LNPs at 4 °C, 22 °C, and 37 °C and found that transfection properties of lyophilized mRNA LNPs were maintained during at least 12 weeks. To our knowledge, this is the first study that demonstrates that optimally lyophilized mRNA LNPs can be safely stored at higher temperatures for months without losing their transfection properties.
MeSH term(s)	Humans ; Temperature ; RNA, Messenger ; COVID-19 Vaccines ; COVID-19 ; Freeze Drying/methods ; Nanoparticles ; Lipids
Chemical Substances	Lipid Nanoparticles ; RNA, Messenger ; COVID-19 Vaccines ; Lipids
Language	English
Publishing date	2023-03-30
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	632533-6
ISSN	1873-4995 ; 0168-3659
ISSN (online)	1873-4995
ISSN	0168-3659
DOI	10.1016/j.jconrel.2023.03.039
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Article: The dawn of mRNA vaccines: The COVID-19 case

Verbeke, Rein / Lentacker, Ine / De Smedt, Stefaan C / Dewitte, Heleen

Journal of controlled release. 2021 May 10, v. 333

2021

Abstract	In less than one year since the outbreak of the COVID-19 pandemic, two mRNA-based vaccines, BNT162b2 and mRNA-1273, were granted the first historic authorization for emergency use, while another mRNA vaccine, CVnCoV, progressed to phase 3 clinical testing. The COVID-19 mRNA vaccines represent a new class of vaccine products, which consist of synthetic mRNA strands encoding the SARS-CoV-2 Spike glycoprotein, packaged in lipid nanoparticles to deliver mRNA to cells. This review digs deeper into the scientific breakthroughs of the last decades that laid the foundations for the rapid rise of mRNA vaccines during the COVID-19 pandemic. As well as providing momentum for mRNA vaccines, SARS-CoV-2 represents an ideal case study allowing to compare design-activity differences between the different mRNA vaccine candidates. Therefore, a detailed overview of the composition and (pre)clinical performance of the three most advanced mRNA vaccines is provided and the influence of choices in their structural design on to their immunogenicity and reactogenicity profile is discussed in depth. In addition to the new fundamental insights in the mRNA vaccines' mode of action highlighted here, we also point out which unknowns remain that require further investigation and possibly, optimization in future mRNA vaccine development.
Keywords	COVID-19 infection ; Severe acute respiratory syndrome coronavirus 2 ; case studies ; glycoproteins ; immunogenicity ; lipids ; mechanism of action ; momentum ; vaccine development ; vaccines
Language	English
Dates of publication	2021-0510
Size	p. 511-520.
Publishing place	Elsevier B.V.
Document type	Article
Note	NAL-light
ZDB-ID	632533-6
ISSN	1873-4995 ; 0168-3659
ISSN (online)	1873-4995
ISSN	0168-3659
DOI	10.1016/j.jconrel.2021.03.043
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Fluorine MR Imaging Probes Dynamic Migratory Profiles of Perfluorocarbon-Loaded Dendritic Cells After Streptozotocin-Induced Inflammation.

Saini, Shweta / Vanherwegen, An-Sofie / Liang, Sayuan / Verbeke, Rein / Korf, Hannelie / Lentacker, Ine / De Smedt, Stefaan C / Gysemans, Conny / Himmelreich, Uwe

Molecular imaging and biology

2022 Volume 24, Issue 2, Page(s) 321–332

Abstract: Purpose: The pathogenesis of type 1 diabetes (T1D) involves presentation of islet-specific self-antigens by dendritic cells (DCs) to autoreactive T cells, resulting in the destruction of insulin-producing pancreatic beta cells. We aimed to study the ... ...

Abstract	Purpose: The pathogenesis of type 1 diabetes (T1D) involves presentation of islet-specific self-antigens by dendritic cells (DCs) to autoreactive T cells, resulting in the destruction of insulin-producing pancreatic beta cells. We aimed to study the dynamic homing of diabetes-prone DCs to the pancreas and nearby organs with and without induction of pancreatic stress in a T1D susceptible model of repeated streptozotocin (STZ) injection. Procedures: In vitro labeling of activated bone marrow-derived DCs (BMDCs) from NOD (Nonobese diabetes) mice was performed using zonyl perfluoro-15-crown-5-ether nanoparticles (ZPFCE-NPs). Internalization of particles was confirmed by confocal microscopy. Two groups of NOD.SCID (nonobese diabetic/severe combined immunodeficiency) mice with (induced by low dose STZ administration) or without pancreatic stress were compared. Diabetogenic BMDCs loaded with BDC2.5 mimotope were pre-labeled with ZPFCE-NPs and adoptively transferred into mice. Longitudinal in vivo fluorine MRI ( Results: In vitro flow cytometry and confocal microscopy confirmed high uptake of nanoparticles in BMDCs during the process of maturation. Migration/homing of activated and ZPFCE-NP- labeled BMDCs to different organs was monitored and quantified longitudinally, showing highest cell density in pancreas at 48-h time-point. Based on Conclusion: We showed the potential of
MeSH term(s)	Animals ; Dendritic Cells ; Diabetes Mellitus, Type 1 ; Fluorine ; Fluorocarbons ; Inflammation ; Magnetic Resonance Imaging/methods ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Streptozocin
Chemical Substances	Fluorocarbons ; Fluorine (284SYP0193) ; Streptozocin (5W494URQ81)
Language	English
Publishing date	2022-01-20
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2079160-4
ISSN	1860-2002 ; 1536-1632
ISSN (online)	1860-2002
ISSN	1536-1632
DOI	10.1007/s11307-021-01701-1
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Article ; Online: Considerations on the Design of Lipid-based mRNA Vaccines Against Cancer.

Meulewaeter, Sofie / Zhang, Yao / Wadhwa, Abishek / Fox, Kevin / Lentacker, Ine / Harder, Kenneth W / Cullis, Pieter R / De Smedt, Stefaan C / Cheng, Miffy H Y / Verbeke, Rein

Journal of molecular biology

2023 Volume 436, Issue 2, Page(s) 168385

Abstract: Throughout the last decades, mRNA vaccines have been developed as a cancer immunotherapeutic and the technology recently gained momentum during the COVID-19 pandemic. Recent promising results obtained from clinical trials investigating lipid-based mRNA ... ...

Abstract	Throughout the last decades, mRNA vaccines have been developed as a cancer immunotherapeutic and the technology recently gained momentum during the COVID-19 pandemic. Recent promising results obtained from clinical trials investigating lipid-based mRNA vaccines in cancer therapy further highlighted the potential of this therapy. Interestingly, while the technologies being used in authorized mRNA vaccines for the prevention of COVID-19 are relatively similar, mRNA vaccines in clinical development for cancer vaccination show marked differences in mRNA modification, lipid carrier, and administration route. In this review, we describe findings on how these factors can impact the potency of mRNA vaccines in cancer therapy and provide insights into the complex interplay between them. We discuss how lipid carrier composition can affect passive targeting to immune cells to improve the efficacy and safety of mRNA vaccines. Finally, we summarize strategies that are established or still being explored to improve the efficacy of mRNA cancer vaccines and include next-generation vaccines that are on the horizon in clinical development.
MeSH term(s)	Humans ; Lipids ; mRNA Vaccines ; Neoplasms/therapy ; Cancer Vaccines ; Vaccine Development/methods
Chemical Substances	Lipids ; mRNA Vaccines ; Cancer Vaccines
Language	English
Publishing date	2023-12-06
Publishing country	Netherlands
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ZDB-ID	80229-3
ISSN	1089-8638 ; 0022-2836
ISSN (online)	1089-8638
ISSN	0022-2836
DOI	10.1016/j.jmb.2023.168385
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Article ; Online: TCR transgenic clone selection guided by immune receptor analysis and single cell RNA expression of polyclonal responders

Debeuf, Nincy / Lameire, Sahine / Vanheerswynghels, Manon / Deckers, Julie / De Wolf, Caroline / Toussaint, Wendy / Verbeke, Rein / Verstaen, Kevin / Hammad, Hamida / Vanhee, Stijn / Lambrecht, Bart N.

bioRxiv

Abstract: Since the precursor frequency of naive T cells is extremely low, investigating the early steps of antigen-specific T cell activation is challenging. To overcome this detection problem, adoptive transfer of a cohort of T cells purified from T cell ... ...

Abstract	Since the precursor frequency of naive T cells is extremely low, investigating the early steps of antigen-specific T cell activation is challenging. To overcome this detection problem, adoptive transfer of a cohort of T cells purified from T cell receptor (TCR) transgenic donors has been extensively used but is not readily available for emerging pathogens. Constructing TCR transgenic mice from T cell hybridomas is a labor-intensive and sometimes erratic process, since the best clones are selected based on antigen-induced CD69 upregulation or IL-2 production in vitro, and TCR chains are PCR-cloned into expression vectors. Here, we exploited the rapid advances in single cell sequencing and TCR repertoire analysis to select the best clones without hybridoma selection, and generated CORSET8 mice (CORona Spike Epitope specific CD8 T cell), carrying a TCR specific for the Spike protein of SARS-CoV-2. Implementing newly created DALI software for TCR repertoire analysis in single cell analysis enabled the rapid selection of the ideal responder CD8 T cell clone, based on antigen reactivity, proliferation and immunophenotype in vivo. In contrast, a traditional method based on hybridoma technology was unsuccessful. Identified TCR sequences were inserted as synthetic DNA into an expression vector and transgenic CORSET8 donor mice were created. After immunization with Spike/CpG-motifs, mRNA vaccination or SARS-CoV2 infection, CORSET8 T cells strongly proliferated and showed signs of T cell activation. Thus, a combination of TCR repertoire analysis and scRNA immunophenotyping allowed rapid selection of antigen-specific TCR sequences that can be used to generate TCR transgenic mice.
Keywords	covid19
Language	English
Publishing date	2024-03-27
Publisher	Cold Spring Harbor Laboratory
Document type	Article ; Online
DOI	10.1101/2024.03.27.586931
Database	COVID19

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Article ; Online: Nanobody-mediated SPECT/CT imaging reveals the spatiotemporal expression of programmed death-ligand 1 in response to a CD8

Ertveldt, Thomas / Meulewaeter, Sofie / De Vlaeminck, Yannick / Olarte, Oscar / Broos, Katrijn / Van Calenbergh, Serge / Bourgeois, Stephanie / Deprez, Joke / Heremans, Yves / Goyvaerts, Cleo / Staels, Willem / De Smedt, Stefaan / Dewitte, Heleen / Devoogdt, Nick / Keyaerts, Marleen / Verbeke, Rein / Barbé, Kurt / Lentacker, Ine / Breckpot, Karine

Theranostics

2023 Volume 13, Issue 15, Page(s) 5483–5500

Abstract: Rationale: ...

Abstract	Rationale:
MeSH term(s)	Humans ; Mice ; Animals ; B7-H1 Antigen ; Natural Killer T-Cells/metabolism ; Single-Domain Antibodies/metabolism ; Immune Checkpoint Inhibitors/metabolism ; Programmed Cell Death 1 Receptor/metabolism ; CD8-Positive T-Lymphocytes ; Tomography, Emission-Computed, Single-Photon ; Tomography, X-Ray Computed ; Vaccines, Synthetic ; Melanoma/diagnostic imaging ; Melanoma/therapy ; Tumor Microenvironment ; mRNA Vaccines
Chemical Substances	CD274 protein, human ; B7-H1 Antigen ; Single-Domain Antibodies ; Immune Checkpoint Inhibitors ; Programmed Cell Death 1 Receptor ; Vaccines, Synthetic
Language	English
Publishing date	2023-10-09
Publishing country	Australia
Document type	Journal Article
ZDB-ID	2592097-2
ISSN	1838-7640 ; 1838-7640
ISSN (online)	1838-7640
ISSN	1838-7640
DOI	10.7150/thno.85106
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