Intratumoral delivery of lipid nanoparticle-formulated mRNA encoding IL-21, IL-7, and 4-1BBL induces systemic anti-tumor immunity

Ahmed E. I. Hamouda; Jessica Filtjens; Elisabeth Brabants; Daliya Kancheva; Ayla Debraekeleer; Jan Brughmans; Lotte Jacobs; Pauline M. R. Bardet; Elisabeth Knetemann; Pierre Lefesvre; Lize Allonsius; Mark Gontsarik; Ismael Varela; Marian Crabbé; Emile J. Clappaert; Federica Cappellesso; Aarushi A. Caro; Alícia Gordún Peiró; Luna Fredericq; Eva Hadadi; Mariona Estapé Senti; Raymond Schiffelers; Leo A. van Grunsven; Frank Aboubakar Nana; Bruno G. De Geest; Sofie Deschoemaeker; Stefaan De Koker; Florence Lambolez; Damya Laoui

doi:10.1038/s41467-024-54877-9

Intratumoral delivery of lipid nanoparticle-formulated mRNA encoding IL-21, IL-7, and 4-1BBL induces systemic anti-tumor immunity

Ahmed E. I. Hamouda, Jessica Filtjens, Elisabeth Brabants, Daliya Kancheva, Ayla Debraekeleer, Jan Brughmans, Lotte Jacobs, Pauline M. R. Bardet, Elisabeth Knetemann, Pierre Lefesvre, Lize Allonsius, Mark Gontsarik, Ismael Varela, Marian Crabbé, Emile J. Clappaert, Federica Cappellesso, Aarushi A. Caro, Alícia Gordún Peiró, Luna Fredericq, Eva HadadiMariona Estapé Senti, Raymond Schiffelers, Leo A. van Grunsven, Frank Aboubakar Nana, Bruno G. De Geest, Sofie Deschoemaeker, Stefaan De Koker, Florence Lambolez, Damya Laoui

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

Abstract

Local delivery of mRNA-based immunotherapy offers a promising avenue as it enables the production of specific immunomodulatory proteins that can stimulate the immune system to recognize and eliminate cancer cells while limiting systemic exposure and toxicities. Here, we develop and employ lipid-based nanoparticles (LNPs) to intratumorally deliver an mRNA mixture encoding the cytokines interleukin (IL)−21 and IL-7 and the immunostimulatory molecule 4-1BB ligand (Triplet LNP). IL-21 synergy with IL-7 and 4-1BBL leads to a profound increase in the frequency of tumor-infiltrating CD8⁺ T cells and their capacity to produce granzyme B and IFN-γ, leading to tumor eradication and the development of long-term immunological memory. Mechanistically, the efficacy of the Triplet LNP depends on tumor-draining lymph nodes to tumor CD8⁺ T-cell trafficking. Moreover, we highlight the therapeutic potential of the Triplet LNP in multiple tumor models in female mice and its superior therapeutic efficacy to immune checkpoint blockade. Ultimately, the expression of these immunomodulators is associated with better overall survival in patients with cancer.

Original language	English
Article number	10635
Number of pages	20
Journal	Nature communications
Volume	15
Issue number	1
DOIs	https://doi.org/10.1038/s41467-024-54877-9
Publication status	Published - 6 Dec 2024

Bibliographical note

Funding Information:
We thank Nadia Abou, Constantinos Papadopoulos, Eleonora Omasta, Ellen Vaneetvelde, and Mait\u00E9 Schuurmans for administrative and technical assistance. We are thankful to the VIB single cell core for providing access to RNA sequencing technologies and the Flow Cytometry Core Facility of Vrije Universiteit Brussel for the use of the Symphony A3 Flow Cytometer. A.E.I.H., J.F., E.B., A.D., L.J., I.V., M.C., S.D.K. and F.L. are supported by a grant from VLAIO (Flanders innovation & entrepreneurship (HBC.2019.2737). J.F., E.B., L.J., I.V., M.C., M.E.S., R.S., S.D.K. and F.L. were supported by the EXPERT project, which has received Funding from the European Union\u2019s Horizon 2020 research and innovation program under Grant Agreement No. 825828. P.M.R.B, L.A., A.A.C. and A.G.P. are supported by predoctoral grants from FWO Vlaanderen (1154720\u2009N, 11P1824N, 1169521\u2009N, 1SH0424N). E.K and LAvG are funded by grants from the FWO and Vrije Universiteit Brussel. M.G. and B.G.D.G. are supported by grants from Stichting tegen kanker and Ghent University. F.C and L.F are supported by Vrije Universiteit Brussel. E.H. is supported by a postdoctoral grant from FWO Vlaanderen (12Y1922N). F.A.N is supported by the Fonds de la Recherche Scientifique FNRS (Belgium) Grants and Fellowships, S.D.S is supported by a postdoctoral grant from Stichting tegen kanker (2021-023). D.L. is supported by grants from FWO, Kom op tegen Kanker, Stichting tegen kanker, VIB and Vrije Universiteit Brussel.

Publisher Copyright:
© The Author(s) 2024.

Access to Document

10.1038/s41467-024-54877-9

Cite this

Hamouda, A. E. I., Filtjens, J., Brabants, E., Kancheva, D., Debraekeleer, A., Brughmans, J., Jacobs, L., Bardet, P. M. R., Knetemann, E., Lefesvre, P., Allonsius, L., Gontsarik, M., Varela, I., Crabbé, M., Clappaert, E. J., Cappellesso, F., Caro, A. A., Peiró, A. G., Fredericq, L., ... Laoui, D. (2024). Intratumoral delivery of lipid nanoparticle-formulated mRNA encoding IL-21, IL-7, and 4-1BBL induces systemic anti-tumor immunity. Nature communications, 15(1), [10635]. https://doi.org/10.1038/s41467-024-54877-9

Hamouda, Ahmed E. I. ; Filtjens, Jessica ; Brabants, Elisabeth ; Kancheva, Daliya ; Debraekeleer, Ayla ; Brughmans, Jan ; Jacobs, Lotte ; Bardet, Pauline M. R. ; Knetemann, Elisabeth ; Lefesvre, Pierre ; Allonsius, Lize ; Gontsarik, Mark ; Varela, Ismael ; Crabbé, Marian ; Clappaert, Emile J. ; Cappellesso, Federica ; Caro, Aarushi A. ; Peiró, Alícia Gordún ; Fredericq, Luna ; Hadadi, Eva ; Senti, Mariona Estapé ; Schiffelers, Raymond ; Grunsven, Leo A. van ; Nana, Frank Aboubakar ; Geest, Bruno G. De ; Deschoemaeker, Sofie ; Koker, Stefaan De ; Lambolez, Florence ; Laoui, Damya. / Intratumoral delivery of lipid nanoparticle-formulated mRNA encoding IL-21, IL-7, and 4-1BBL induces systemic anti-tumor immunity. In: Nature communications. 2024 ; Vol. 15, No. 1.

@article{3d68dfd275cd4584ab22eaa9f74e32c5,

title = "Intratumoral delivery of lipid nanoparticle-formulated mRNA encoding IL-21, IL-7, and 4-1BBL induces systemic anti-tumor immunity",

abstract = "Local delivery of mRNA-based immunotherapy offers a promising avenue as it enables the production of specific immunomodulatory proteins that can stimulate the immune system to recognize and eliminate cancer cells while limiting systemic exposure and toxicities. Here, we develop and employ lipid-based nanoparticles (LNPs) to intratumorally deliver an mRNA mixture encoding the cytokines interleukin (IL)−21 and IL-7 and the immunostimulatory molecule 4-1BB ligand (Triplet LNP). IL-21 synergy with IL-7 and 4-1BBL leads to a profound increase in the frequency of tumor-infiltrating CD8+ T cells and their capacity to produce granzyme B and IFN-γ, leading to tumor eradication and the development of long-term immunological memory. Mechanistically, the efficacy of the Triplet LNP depends on tumor-draining lymph nodes to tumor CD8+ T-cell trafficking. Moreover, we highlight the therapeutic potential of the Triplet LNP in multiple tumor models in female mice and its superior therapeutic efficacy to immune checkpoint blockade. Ultimately, the expression of these immunomodulators is associated with better overall survival in patients with cancer.",

author = "Hamouda, {Ahmed E. I.} and Jessica Filtjens and Elisabeth Brabants and Daliya Kancheva and Ayla Debraekeleer and Jan Brughmans and Lotte Jacobs and Bardet, {Pauline M. R.} and Elisabeth Knetemann and Pierre Lefesvre and Lize Allonsius and Mark Gontsarik and Ismael Varela and Marian Crabb{\'e} and Clappaert, {Emile J.} and Federica Cappellesso and Caro, {Aarushi A.} and Peir{\'o}, {Al{\'i}cia Gord{\'u}n} and Luna Fredericq and Eva Hadadi and Senti, {Mariona Estap{\'e}} and Raymond Schiffelers and Grunsven, {Leo A. van} and Nana, {Frank Aboubakar} and Geest, {Bruno G. De} and Sofie Deschoemaeker and Koker, {Stefaan De} and Florence Lambolez and Damya Laoui",

note = "Funding Information: We thank Nadia Abou, Constantinos Papadopoulos, Eleonora Omasta, Ellen Vaneetvelde, and Mait\u00E9 Schuurmans for administrative and technical assistance. We are thankful to the VIB single cell core for providing access to RNA sequencing technologies and the Flow Cytometry Core Facility of Vrije Universiteit Brussel for the use of the Symphony A3 Flow Cytometer. A.E.I.H., J.F., E.B., A.D., L.J., I.V., M.C., S.D.K. and F.L. are supported by a grant from VLAIO (Flanders innovation & entrepreneurship (HBC.2019.2737). J.F., E.B., L.J., I.V., M.C., M.E.S., R.S., S.D.K. and F.L. were supported by the EXPERT project, which has received Funding from the European Union\u2019s Horizon 2020 research and innovation program under Grant Agreement No. 825828. P.M.R.B, L.A., A.A.C. and A.G.P. are supported by predoctoral grants from FWO Vlaanderen (1154720\u2009N, 11P1824N, 1169521\u2009N, 1SH0424N). E.K and LAvG are funded by grants from the FWO and Vrije Universiteit Brussel. M.G. and B.G.D.G. are supported by grants from Stichting tegen kanker and Ghent University. F.C and L.F are supported by Vrije Universiteit Brussel. E.H. is supported by a postdoctoral grant from FWO Vlaanderen (12Y1922N). F.A.N is supported by the Fonds de la Recherche Scientifique FNRS (Belgium) Grants and Fellowships, S.D.S is supported by a postdoctoral grant from Stichting tegen kanker (2021-023). D.L. is supported by grants from FWO, Kom op tegen Kanker, Stichting tegen kanker, VIB and Vrije Universiteit Brussel. Publisher Copyright: {\textcopyright} The Author(s) 2024.",

year = "2024",

month = dec,

day = "6",

doi = "10.1038/s41467-024-54877-9",

language = "English",

volume = "15",

journal = "Nature Communications",

issn = "2041-1723",

publisher = "Nature Publishing Group",

number = "1",

}

Hamouda, AEI, Filtjens, J, Brabants, E, Kancheva, D , Debraekeleer, A , Brughmans, J, Jacobs, L, Bardet, PMR , Knetemann, E , Lefesvre, P , Allonsius, L, Gontsarik, M, Varela, I, Crabbé, M, Clappaert, EJ, Cappellesso, F , Caro, AA , Peiró, AG , Fredericq, L , Hadadi, E, Senti, ME, Schiffelers, R, Grunsven, LAV, Nana, FA, Geest, BGD, Deschoemaeker, S, Koker, SD, Lambolez, F & Laoui, D 2024, 'Intratumoral delivery of lipid nanoparticle-formulated mRNA encoding IL-21, IL-7, and 4-1BBL induces systemic anti-tumor immunity', Nature communications, vol. 15, no. 1, 10635. https://doi.org/10.1038/s41467-024-54877-9

Intratumoral delivery of lipid nanoparticle-formulated mRNA encoding IL-21, IL-7, and 4-1BBL induces systemic anti-tumor immunity. / Hamouda, Ahmed E. I.; Filtjens, Jessica; Brabants, Elisabeth; Kancheva, Daliya ; Debraekeleer, Ayla ; Brughmans, Jan; Jacobs, Lotte; Bardet, Pauline M. R.; Knetemann, Elisabeth ; Lefesvre, Pierre ; Allonsius, Lize; Gontsarik, Mark; Varela, Ismael; Crabbé, Marian; Clappaert, Emile J.; Cappellesso, Federica ; Caro, Aarushi A.; Peiró, Alícia Gordún ; Fredericq, Luna ; Hadadi, Eva; Senti, Mariona Estapé; Schiffelers, Raymond; Grunsven, Leo A. van; Nana, Frank Aboubakar; Geest, Bruno G. De; Deschoemaeker, Sofie; Koker, Stefaan De; Lambolez, Florence; Laoui, Damya.

In: Nature communications, Vol. 15, No. 1, 10635, 06.12.2024.

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

TY - JOUR

T1 - Intratumoral delivery of lipid nanoparticle-formulated mRNA encoding IL-21, IL-7, and 4-1BBL induces systemic anti-tumor immunity

AU - Hamouda, Ahmed E. I.

AU - Filtjens, Jessica

AU - Brabants, Elisabeth

AU - Kancheva, Daliya

AU - Debraekeleer, Ayla

AU - Brughmans, Jan

AU - Jacobs, Lotte

AU - Bardet, Pauline M. R.

AU - Knetemann, Elisabeth

AU - Lefesvre, Pierre

AU - Allonsius, Lize

AU - Gontsarik, Mark

AU - Varela, Ismael

AU - Crabbé, Marian

AU - Clappaert, Emile J.

AU - Cappellesso, Federica

AU - Caro, Aarushi A.

AU - Peiró, Alícia Gordún

AU - Fredericq, Luna

AU - Hadadi, Eva

AU - Senti, Mariona Estapé

AU - Schiffelers, Raymond

AU - Grunsven, Leo A. van

AU - Nana, Frank Aboubakar

AU - Geest, Bruno G. De

AU - Deschoemaeker, Sofie

AU - Koker, Stefaan De

AU - Lambolez, Florence

AU - Laoui, Damya

N1 - Funding Information: We thank Nadia Abou, Constantinos Papadopoulos, Eleonora Omasta, Ellen Vaneetvelde, and Mait\u00E9 Schuurmans for administrative and technical assistance. We are thankful to the VIB single cell core for providing access to RNA sequencing technologies and the Flow Cytometry Core Facility of Vrije Universiteit Brussel for the use of the Symphony A3 Flow Cytometer. A.E.I.H., J.F., E.B., A.D., L.J., I.V., M.C., S.D.K. and F.L. are supported by a grant from VLAIO (Flanders innovation & entrepreneurship (HBC.2019.2737). J.F., E.B., L.J., I.V., M.C., M.E.S., R.S., S.D.K. and F.L. were supported by the EXPERT project, which has received Funding from the European Union\u2019s Horizon 2020 research and innovation program under Grant Agreement No. 825828. P.M.R.B, L.A., A.A.C. and A.G.P. are supported by predoctoral grants from FWO Vlaanderen (1154720\u2009N, 11P1824N, 1169521\u2009N, 1SH0424N). E.K and LAvG are funded by grants from the FWO and Vrije Universiteit Brussel. M.G. and B.G.D.G. are supported by grants from Stichting tegen kanker and Ghent University. F.C and L.F are supported by Vrije Universiteit Brussel. E.H. is supported by a postdoctoral grant from FWO Vlaanderen (12Y1922N). F.A.N is supported by the Fonds de la Recherche Scientifique FNRS (Belgium) Grants and Fellowships, S.D.S is supported by a postdoctoral grant from Stichting tegen kanker (2021-023). D.L. is supported by grants from FWO, Kom op tegen Kanker, Stichting tegen kanker, VIB and Vrije Universiteit Brussel. Publisher Copyright: © The Author(s) 2024.

PY - 2024/12/6

Y1 - 2024/12/6

N2 - Local delivery of mRNA-based immunotherapy offers a promising avenue as it enables the production of specific immunomodulatory proteins that can stimulate the immune system to recognize and eliminate cancer cells while limiting systemic exposure and toxicities. Here, we develop and employ lipid-based nanoparticles (LNPs) to intratumorally deliver an mRNA mixture encoding the cytokines interleukin (IL)−21 and IL-7 and the immunostimulatory molecule 4-1BB ligand (Triplet LNP). IL-21 synergy with IL-7 and 4-1BBL leads to a profound increase in the frequency of tumor-infiltrating CD8+ T cells and their capacity to produce granzyme B and IFN-γ, leading to tumor eradication and the development of long-term immunological memory. Mechanistically, the efficacy of the Triplet LNP depends on tumor-draining lymph nodes to tumor CD8+ T-cell trafficking. Moreover, we highlight the therapeutic potential of the Triplet LNP in multiple tumor models in female mice and its superior therapeutic efficacy to immune checkpoint blockade. Ultimately, the expression of these immunomodulators is associated with better overall survival in patients with cancer.

AB - Local delivery of mRNA-based immunotherapy offers a promising avenue as it enables the production of specific immunomodulatory proteins that can stimulate the immune system to recognize and eliminate cancer cells while limiting systemic exposure and toxicities. Here, we develop and employ lipid-based nanoparticles (LNPs) to intratumorally deliver an mRNA mixture encoding the cytokines interleukin (IL)−21 and IL-7 and the immunostimulatory molecule 4-1BB ligand (Triplet LNP). IL-21 synergy with IL-7 and 4-1BBL leads to a profound increase in the frequency of tumor-infiltrating CD8+ T cells and their capacity to produce granzyme B and IFN-γ, leading to tumor eradication and the development of long-term immunological memory. Mechanistically, the efficacy of the Triplet LNP depends on tumor-draining lymph nodes to tumor CD8+ T-cell trafficking. Moreover, we highlight the therapeutic potential of the Triplet LNP in multiple tumor models in female mice and its superior therapeutic efficacy to immune checkpoint blockade. Ultimately, the expression of these immunomodulators is associated with better overall survival in patients with cancer.

UR - https://doi.org/10.1038/s41467-024-54877-9

UR - http://www.scopus.com/inward/record.url?scp=85211387402&partnerID=8YFLogxK

U2 - 10.1038/s41467-024-54877-9

DO - 10.1038/s41467-024-54877-9

M3 - Article

C2 - 39639025

VL - 15

JO - Nature Communications

JF - Nature Communications

SN - 2041-1723

IS - 1

M1 - 10635

ER -

Intratumoral delivery of lipid nanoparticle-formulated mRNA encoding IL-21, IL-7, and 4-1BBL induces systemic anti-tumor immunity

Abstract

Bibliographical note

Access to Document

Other files and links

Cite this