Direct Injection of Protamine-protected mRNA: Results of a Phase 1/2 Vaccination Trial in Metastatic Melanoma Patients

Weide, Benjamin; Pascolo, Steve; Scheel, Birgit; Derhovanessian, Evelyna; Pflugfelder, Annette; Eigentler, Thomas; Pawelec, Graham; Hoerr, Ingmar; Rammensee, Hans‐Georg; Garbe, Claus

doi:10.1097/cji.0b013e3181a00068

Cited by 309 publications

(233 citation statements)

References 24 publications

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“…Successful applications of RNA nanoparticle-based vaccines that are independent of ex vivo transfection of antigenpresenting cells are limited in animal models (reviewed in 32,93), and few such approaches have made it to clinical trials. Although correlates of immune protection in humans have been reported, clinical efficacy has been disappointing (94)(95)(96)(97). Replicons based on RNA viruses of the alphavirus family, such as SFV, VEEV, and Sindbis virus (SIN), have served as vaccine vectors, usually delivered as replication-deficient pseudoviral particles generated through complementation of structural genes in cell culture (98).…”

Section: Discussionmentioning

confidence: 99%

Dendrimer-RNA nanoparticles generate protective immunity against lethal Ebola, H1N1 influenza, and Toxoplasma gondii challenges with a single dose

Chahal

Khan

Cooper

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

329

260

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Vaccines have had broad medical impact, but existing vaccine technologies and production methods are limited in their ability to respond rapidly to evolving and emerging pathogens, or sudden outbreaks. Here, we develop a rapid-response, fully synthetic, single-dose, adjuvant-free dendrimer nanoparticle vaccine platform wherein antigens are encoded by encapsulated mRNA replicons. To our knowledge, this system is the first capable of generating protective immunity against a broad spectrum of lethal pathogen challenges, including H1N1 influenza, Toxoplasma gondii, and Ebola virus. The vaccine can be formed with multiple antigen-expressing replicons, and is capable of eliciting both CD8+ T-cell and antibody responses. The ability to generate viable, contaminant-free vaccines within days, to single or multiple antigens, may have broad utility for a range of diseases.

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Section: Discussionmentioning

confidence: 99%

Dendrimer-RNA nanoparticles generate protective immunity against lethal Ebola, H1N1 influenza, and Toxoplasma gondii challenges with a single dose

Chahal

Khan

Cooper

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

329

260

View full text Add to dashboard Cite

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“…The reason, however, why we are excited regarding the fact that in immature DCs b-S-ARCA(D1) cap analog profoundly enhances RNA stability and translational efficiency, is our particular interest in using the RNA naked and for direct injection, for example, intradermally or into the lymph node for uptake by local DCs. [7][8][9] In summary, our data reveal 5 0 mRNA cap analogs as a further structural feature to modify the stability and translational efficiency of in vitro transcribed RNA for pharmaceutical applications. Together with other modifications as previously described, 13,14 we now have a tool-box in hand to design RNAs with properties suited for a diversity of applications, further enhancing the potential of RNA as a promising drug compound.…”

Section: Discussionmentioning

confidence: 99%

“…4,5 Preclinical experiments and human clinical trials testing administration of autologous DCs loaded in vitro with RNA as well as direct injection of RNA via different routes to reach DCs in situ showed feasibility, lack of toxicity and induction of the expected antigen-specific immune response. [6][7][8][9][10][11] Improvement of immunobioavailability of RNA-based vaccines in DCs has been a recurrent subject of our research, because the dose of the antigen may be one of the critical factors for generating strong and sustained antigen-specific immune responses. 12 In previous work, we have developed plasmid templates for in vitro transcription of RNA-encoded antigen with modified 3 0 structures (3 0 UTR and poly(A)tail) stabilizing the RNA and optimizing its translational performance.…”

Section: Introductionmentioning

confidence: 99%

Phosphorothioate cap analogs increase stability and translational efficiency of RNA vaccines in immature dendritic cells and induce superior immune responses in vivo

et al. 2010

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Vaccination with in vitro transcribed RNA coding for tumor antigens is considered a promising approach for cancer immunotherapy and has already entered human clinical testing. One of the basic objectives for development of RNA as a drug is the optimization of immunobioavailability of the encoded antigen in vivo. By analyzing the effect of different synthetic 5 0 mRNA cap analogs on the kinetics of the encoded protein, we found that m 2 7,2 0 ÀO Gpp S pG (b-S-ARCA) phosphorothioate caps, in particular the D1 diastereoisomer, profoundly enhance RNA stability and translational efficiency in immature but not mature dendritic cells. Moreover, in vivo delivery of the antigen as b-S-ARCA(D1)-capped RNA species is superior for protein expression and for efficient priming and expansion of naïve antigen-specific T cells in mice. Our findings establish 5 0 mRNA cap analogs as yet another module for tuning immunopharmacological properties of recombinant antigen-encoding RNA for vaccination purposes.

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“…[5][6][7] The other employs direct injection of naked RNA. 8,9 Feasibility and safety of vaccination protocols based on intradermal administration of naked RNA have been shown in preclinical and clinical settings and partial protection from tumor challenge has been achieved in mice. 10,11 However, at present, neither in animals nor in men, systemic immune responses let alone therapeutic effects were unequivocally demonstrated.…”

mentioning

confidence: 99%

Selective uptake of naked vaccine RNA by dendritic cells is driven by macropinocytosis and abrogated upon DC maturation

et al. 2011

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Even though it is known for more than one decade that antigen-encoding RNA can deliver antigenic information to induce antigen-specific immunity against cancer, the nature and mechanism of RNA uptake have remained enigmatic. In this study, we investigated the pharmacokinetics of naked RNA administered into the lymph node. We observed that RNA is rapidly and selectively uptaken by lymph node dendritic cells (DCs). Furthermore, in vitro and in vivo studies revealed that the efficient internalization of RNA by human and murine DCs is primarily driven by macropinocytosis. Selective inhibition of macropinocytosis by compounds or as a consequence of DC maturation abrogated RNA internalization and delivery of encoded antigens. Our findings imply that bioavailability of recombinant RNA vaccines in vivo highly depends on the density and the maturation stage of DCs at the administration site and are of importance for the design of RNA-based clinical immunotherapy protocols. Gene Therapy (2011) Keywords: vaccination; dendritic cells; RNA; macropinocytosis INTRODUCTION Antigen-encoding RNA has emerged as an attractive new vaccine format. Major advantages of RNA are efficient delivery of the complete antigenic information, transient expression and lack of genome integration, as well as cost-efficient and easy production in large amounts and high purity. 1 Moreover, through activation of the immune system via TLR3, TLR7 and TLR8, 2,3 RNA supports the induction of immune responses against the encoded protein.Of several in vitro and in vivo strategies utilizing antigen-encoding RNA as a vaccine format, 4 two are in clinical stage. One is based on adoptive transfer of autologous dendritic cells (DCs) transfected ex vivo with antigen-encoding RNA. [5][6][7] The other employs direct injection of naked RNA. 8,9 Feasibility and safety of vaccination protocols based on intradermal administration of naked RNA have been shown in preclinical and clinical settings and partial protection from tumor challenge has been achieved in mice. 10,11 However, at present, neither in animals nor in men, systemic immune responses let alone therapeutic effects were unequivocally demonstrated. It is assumed that rapid degradation by ubiquitous RNAses constraining pharmacologically efficient dosing of unprotected RNA contributes to the still suboptimal clinical effects. Knowledge about the nature and pharmacokinetics of RNA uptake in vitro and in vivo as well as the cell-type specificity is still scarce. Recently, we discovered that the administration route of naked RNA has a crucial impact on its efficacy as a vaccine. By injecting RNA directly into lymph nodes of mice, we achieved for the first time strong systemic antigen-specific Th1 type immunity and cancer cure in preclinical animal models. 12

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Direct Injection of Protamine-protected mRNA: Results of a Phase 1/2 Vaccination Trial in Metastatic Melanoma Patients

Cited by 309 publications

References 24 publications

Dendrimer-RNA nanoparticles generate protective immunity against lethal Ebola, H1N1 influenza, and Toxoplasma gondii challenges with a single dose

Dendrimer-RNA nanoparticles generate protective immunity against lethal Ebola, H1N1 influenza, and Toxoplasma gondii challenges with a single dose

Phosphorothioate cap analogs increase stability and translational efficiency of RNA vaccines in immature dendritic cells and induce superior immune responses in vivo

Selective uptake of naked vaccine RNA by dendritic cells is driven by macropinocytosis and abrogated upon DC maturation

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