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The blood-brain barrier (BBB) restricts the systemic delivery of messenger RNAs (mRNAs) into diseased neurons. Although leucocyte-derived extracellular vesicles (EVs) can cross the BBB at inflammatory sites, it is difficult to efficiently load long mRNAs into the EVs and to enhance their neuronal uptake. Here we show that the packaging of mRNA into leucocyte-derived EVs and the endocytosis of the EVs by neurons can be enhanced by engineering leucocytes to produce EVs that incorporate retrovirus-like mRNA-packaging capsids. We transfected immortalized and primary bone-marrow-derived leucocytes with DNA or RNA encoding the capsid-forming activity-regulated cytoskeleton-associated (Arc) protein as well as capsid-stabilizing Arc 5'-untranslated-region RNA elements. These engineered EVs inherit endothelial adhesion molecules from donor leukocytes, recruit endogenous enveloping proteins to their surface, cross the BBB, and enter the neurons in neuro-inflammatory sites. Produced from self-derived donor leukocytes, the EVs are immunologically inert, and enhanced the neuronal uptake of the packaged mRNA in a mouse model of low-grade chronic neuro-inflammation.
Assuntos
Barreira Hematoencefálica , Vesículas Extracelulares , Neurônios , RNA Mensageiro , Animais , Neurônios/metabolismo , Vesículas Extracelulares/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Camundongos , Barreira Hematoencefálica/metabolismo , Retroviridae/genética , Capsídeo/metabolismo , Leucócitos/metabolismo , Humanos , Camundongos Endogâmicos C57BLRESUMO
The clustered, regularly interspaced, short palindromic repeats (CRISPR)-associated protein 9 (Cas9) system is a powerful genome-editing tool that is widely used in many different applications. However, the high-frequency mutations induced by RNA-guided Cas9 at sites other than the intended on-target sites are a major concern that impedes therapeutic and clinical applications. A deeper analysis shows that most off-target events result from the non-specific mismatch between single guide RNA (sgRNA) and target DNA. Therefore, minimizing the non-specific RNA-DNA interaction can be an effective solution to this issue. Here we provide two novel methods at the protein and mRNA levels to minimize this mismatch issue by chemically conjugating Cas9 with zwitterionic pCB polymers or genetically fusing Cas9 with zwitterionic (EK)n peptides. The zwitterlated or EKylated CRISPR/Cas9 ribonucleoproteins (RNPs) show reduced off-target DNA editing while maintaining a similar level of on-target gene editing activity. Results show that the off-target efficiency of zwitterlated CRISPR/Cas9 is reduced on average by 70% and can be as high as 90% when compared with naive CRISPR/Cas9 editing. These approaches provide a simple and effective way to streamline the development of genome editing with the potential to accelerate a wide array of biological and therapeutic applications based on CRISPR/Cas9 technology.
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Antibodies against poly(ethylene glycol) (PEG) have been found to be the culprit of side reactions and efficacy loss of a number of PEGylated drugs. Fundamental mechanisms of PEG immunogenicity and design principles for PEG alternatives still have not been fully explored. By using hydrophobic interaction chromatography (HIC) under varied salt conditions, we reveal the "hidden" hydrophobicity of those polymers which are generally considered as hydrophilic. A correlation between the hidden hydrophobicity of a polymer and its polymer immunogenicity is observed when this polymer is conjugated with an immunogenic protein. Such a correlation of hidden hydrophobicity vs. immunogenicity for a polymer also applies to corresponding polymer-protein conjugates. Atomistic molecular dynamics (MD) simulation results show a similar trend. Based on polyzwitterion modification and with this HIC technique, we are able to produce extremely low-immunogenic protein conjugates as their hydrophilicity is pushed to the limit and their hydrophobicity is eliminated, breaking the current barriers of eliminating anti-drug and anti-polymer antibodies.
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Although recombinant adeno-associated viruses (AAVs) are considered low immunogenic and safe for gene delivery, the immunogenicity of capsids still represents a major obstacle to the readministration of AAV vectors. Here, we design an immunosuppressive zwitterionic phosphoserine (PS)-containing polypeptide to induce AAV-specific immune tolerance and eradicate the immunological response. AAVs modified with the zwitterionic PS polypeptide maintain their transduction activity and tissue tropism but suppress the induction of AAV-specific antibodies. In a hemophilia A mouse model (FVIII knockout mice), the readministration of zwitterionic PS polypeptide-modified AAV8-FVIII vectors successfully evades immunological response, corrects blood FVIII levels, and stops blood loss in tail-bleeding experiments. This potent and safe technology mimics the natural tolerance of apoptotic cells and controls the immunosuppressive, zwitterionic, and degradable polypeptide precisely, reducing the concern of toxicities upon readministrations. This work presents a new concept and a platform of engineered viral vectors by chemically linking immunosuppressive materials to AAV vectors, enabling the readministration of AAV vectors while maintaining their transduction efficiency to a considerable degree.
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Dependovirus , Terapia Genética , Animais , Camundongos , Fosfosserina , Dependovirus/genética , Vetores Genéticos , Camundongos Knockout , Peptídeos/genéticaRESUMO
Secondary lymphoid organs (SLOs) are an important target for mRNA delivery in various applications. While the current delivery method relies on the drainage of nanoparticles to lymph nodes by intramuscular (IM) or subcutaneous (SC) injections, an efficient mRNA delivery carrier for SLOs-targeting delivery by systemic administration (IV) is highly desirable but yet to be available. In this study, we developed an efficient SLOs-targeting carrier using phosphatidylserine (PS), a well-known signaling molecule that promotes the endocytic activity of phagocytes and cellular entry of enveloped viruses. We adopted these biomimetic strategies and added PS into the standard four-component MC3-based LNP formulation (PS-LNP) to facilitate the cellular uptake of immune cells beyond the charge-driven targeting principle commonly used today. As a result, PS-LNP performed efficient protein expression in both lymph nodes and the spleen after IV administration. In vitro and in vivo characterizations on PS-LNP demonstrated a monocyte/macrophage-mediated SLOs-targeting delivery mechanism.
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Nanopartículas , Fosfatidilserinas , Nanopartículas/química , RNA Interferente Pequeno/genética , RNA Mensageiro/genéticaRESUMO
The design of three-dimensional crosslinked units with a spatial structure is of great significance for improving the mechanical properties of hydrogels. However, almost all the nanocomposites incorporated in hydrogels were defined as rigid nanofillers without further discussion on the potential contribution from the spatial structure change. In this work, the 3D nano chemical crosslinker multilayer graphene oxide acrylate (mGOa) was developed as a pressure-responsive crosslinker to achieve both low elastic modulus and high compression stress by synergizing more polymer chains against the loading force through interlayer sliding. Results showed that the hydrogel crosslinked by only 2 mg/mL mGOa nano chemical crosslinker in the poly(2-hydroxyethyl methacrylate-co-acrylamide) hydrogel (molar ratio: 1:1) can effectively enhance the mechanical strength up to 14.1 ± 2.1 MPa at a high compressive strain (90.6%) with an elastic modulus of less than 0.03 MPa at the initial 5% compression, whereas the hydrogel crosslinked by methacrylated single-layer graphene oxide (sGOa) or a small-molecule chemical crosslinker, N,N'-methylene bisacrylamide, can only reach 2.3 ± 0.8 MPa and 1.4 ± 0.4 MPa, respectively. In addition, the instantaneous modulus of the mGOa crosslinked hydrogel rapidly increased to the peak value with the increase of strain. The repeated compression test of HcA-mGOa hydrogels showed the responsive increase of the modulus, which was promoted by the synergism of polymer chains under compression. This indicated that the interlayer sliding of mGOa is the key contributor to mechanical strength enhancement, which provides a new rationale to design tough hydrogels.
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Zwitterionic hydrogels have received great attention due to their excellent nonfouling and biocompatible properties, but they suffer from weak mechanical strength in the saline environments important for biomedical and engineering applications due to the "anti-polyelectrolyte" effect. Conventional strategies to introduce hydrophobic or non-zwitterionic components to increase mechanical strength compromise their nonfouling properties. Here, a highly effective strategy is reported to achieve both high mechanical strength and excellent nonfouling properties by constructing a pure zwitterionic triple-network (ZTN) hydrogel. The strong electrostatic interaction and network entanglement within the triple-network structure can effectively dissipate energy to toughen the hydrogel and achieve high strength, toughness, and stiffness in saline environments (compressive fracture stress 18.2 ± 1.4 MPa, toughness 1.62 ± 0.03 MJ m-3 , and modulus 0.66 ± 0.03 MPa in seawater environments). Moreover, the ZTN hydrogel is shown to strongly resist the attachment of proteins, bacteria, and cells. The results provide a fundamental understanding to guide the design of tough nonfouling zwitterionic hydrogels for a broad range of applications.
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Inspired by the amino acid composition of natural protein surfaces, we developed a zwitterionic cloak containing multi-layers of short alternating glutamic acid and lysine (EK) peptides as a facile, highly effective and low-immunogenicity approach for the protection and delivery of biotherapeutics. Each EK layer grafted to proteins provides multiple times of new lysine reaction sites for the growth of subsequent EK layers. This unique design allows EK peptides to achieve high coating density on proteins, overcoming the limitation of traditional conjugation strategies that rely on the number of innate lysine groups. A triple-layer EK cloak manifests to successfully eliminate the specific and non-specific interactions of protected asparaginase with biological media while prolong the drug circulation time and significantly mitigate its immunogenicity in vivo, suggesting an EK peptide cloak as a promising approach to improve the safety and efficacy of biotherapeutics.
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Ácido Glutâmico/química , Lisina/química , Peptídeos/química , Proteínas/química , Propriedades de SuperfícieRESUMO
Superhydrophilic zwitterionic polymers are a class of nonfouling materials capable of effectively resisting any nonspecific interactions with biological systems. We designed here a functional zwitterionic polymer that achieves a trade-off between nonspecific interactions providing the nonfouling property and a specific interaction for bioactive functionality. Built from phosphoserine, an immune-signaling molecule in nature, this zwitterionic polymer exhibits both nonfouling and immunomodulatory properties. Its conjugation to uricase is shown to proactively eradicate all unwanted immune response, outperforming the zwitterionic polymers. On the other hand, this polymer could significantly prolong the half-life of protein drugs in vivo, overcoming the innate drawback of phosphoserine in inducing accelerated clearance. Our demonstration of a nonfouling zwitterionic material with built-in immunomodulatory functionality provides new insights into the fundamental design of biomaterials, as well as far-reaching implications for broad applications such as drug delivery, implants, and cell therapy.
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Glucagon-like peptide-1 (GLP-1) is of particular interest for treating type 2 diabetes mellitus (T2DM), as it induces insulin secretion in a glucose-dependent fashion and has the potential to facilitate weight control. However, native GLP-1 is a short incretin peptide that is susceptible to fast proteolytic inactivation and rapid clearance from the circulation. Various GLP-1 analogs and bioconjugation of GLP-1 analogs have been developed to counter these issues, but these modifications are frequently accompanied by the sacrifice of potency and the induction of immunogenicity. Here, we demonstrated that with the conjugation of a zwitterionic polymer, poly(carboxybetaine) (pCB), the pharmacokinetic properties of native GLP-1 were greatly enhanced without serious negative effects on its potency and secondary structure. The pCB conjugated GLP-1 further provided glycemic control for up to 6 days in a mouse study. These results illustrate that the conjugation of pCB could realize the potential of using native GLP-1 for prolonged glycemic control in treating T2DM.
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Diabetes Mellitus Tipo 2/sangue , Peptídeo 1 Semelhante ao Glucagon/química , Controle Glicêmico/métodos , Hipoglicemiantes/uso terapêutico , Polímeros/química , Animais , Diabetes Mellitus Tipo 2/tratamento farmacológico , Modelos Animais de Doenças , Peptídeo 1 Semelhante ao Glucagon/farmacocinética , Peptídeo 1 Semelhante ao Glucagon/uso terapêutico , Meia-Vida , Hipoglicemiantes/farmacocinética , Camundongos , Estrutura Secundária de ProteínaRESUMO
The lymphatic system provides a major route for the dissemination of many diseases such as tumor metastasis and virus infection. At present, treating these diseases remains a knotty task due to the difficulty of delivering sufficient drugs into lymphatics. After subcutaneous (SC) injection, the transferring of drugs to lymphatic vessels is significantly attenuated by physiological barriers in the interstitial space. Moreover, SC injection represents a highly challenging administration route for biological drugs, as it increases the risk of undesirable immune responses. Here, we demonstrate a simple and effective strategy to address this dilemma by conjugating protein therapeutics with zwitterionic poly(carboxy betaine) (PCB) polymers. PCB conjugation to l-asparaginase (ASP), a highly immunogenic enzyme drug, manifests to significantly promote the diffusion of ASP into the lymphatic system while mitigating its immunogenicity. This platform will facilitate the development of new therapies against diverse lymph-related diseases by enabling safe and efficient lymphatic drug delivery.
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Sistemas de Liberação de Medicamentos , Vasos Linfáticos , Nanoconjugados , Preparações Farmacêuticas , Sistema LinfáticoRESUMO
Pulmonary delivery of protein drugs into the systemic circulation is highly desirable as the lung provides a large absorption surface area and a more favorable environment for biologics compared to other delivery routes. However, pulmonary systemic delivery of proteins presents several challenges such as poor protein stability and limited bioavailability, especially for large proteins (molecular weight > 50 kDa), which exhibit an average bioavailability of 1% to 5% when delivered via the pulmonary route. Here, we demonstrated that with the conjugation of zwitterionic poly(carboxybetaine) (pCB) polymer, the bioavailability of organophosphate hydrolase (OPH) was significantly increased from 5% to 53%. OPH conjugated with pCB delivered through intubation-assisted intratracheal instillation (IAIS) into the lung exhibited improved pharmacokinetic properties and prophylactic efficacy against organophosphate poisoning, showing its application potential. Zwitterionic polymer conjugation provides the possibility for a favorable, non-invasive delivery of biological therapeutics into the systemic circulation.
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Preparações Farmacêuticas , Polímeros , Pulmão , Estabilidade Proteica , ProteínasRESUMO
The therapeutic potential of protein drugs has been hindered by difficulties with long-term stability and rapid clearance from the body. Recombinant fusion proteins provide a scalable platform for engineered biologics, whereby a polypeptide domain is appended to alter the physical characteristics of a therapeutic protein and enhance its pharmaceutical viability. Two simple design principles for recombinant fusion proteins, based on the physical properties of the polypeptide domain, have been separately applied to address issues with the stability and delivery of biologics. "Conformationally disordered" peptides, exemplified by the homo amino acid peptide polyG, have been shown to increase the circulation half-life and bioactivity of protein therapeutics in vivo. Superhydrophilic peptides, exemplified by the alternating-charge peptide poly(EK), have been shown to increase the thermostability of proteins in vitro. The combination of superhydrophilicity and conformational disorder in a single fusion peptide could simultaneously address concerns regarding the stability and therapeutic lifetime of biologics. In the current work, we use enhanced sampling molecular dynamics (MD) simulations to investigate the conformational ensemble of poly(EK) and glycine-substituted poly(EK) variants and validate our structural predictions with circular dichroism (CD). We find the (EK)15 peptide exhibits a high propensity for forming antiparallel ß-strand secondary structures, which are stabilized by extensive salt bridging of the positive and negative side chains. MD simulations predict that limited glycine substitutions effectively disrupt the secondary structure and promote disordered conformations at physiologically relevant temperatures. We conclude that the conformational disorder of alternating-charge peptides should be taken into account to improve their suitability for drug delivery applications. We also contribute a computational approach to quantify conformational disorder in polypeptides, which should facilitate the de novo design of effective fusion proteins.
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Peptídeos/química , Engenharia de Proteínas/métodos , Dicroísmo Circular , Glicina/química , Simulação de Dinâmica Molecular , Mutação , Peptídeos/genética , Estrutura Secundária de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , SoluçõesRESUMO
Materials that resist nonspecific protein adsorption are needed for many applications. However, few are able to achieve ultralow fouling in complex biological milieu. Zwitterionic polymers emerge as a class of highly effective ultralow fouling materials due to their superhydrophilicity, outperforming other hydrophilic materials such as poly(ethylene glycol). Unfortunately, there are only three major classes of zwitterionic materials based on poly(phosphorylcholine), poly(sulfobetaine), and poly(carboxybetaine) currently available. Inspired by trimethylamine N-oxide (TMAO), a zwitterionic osmolyte and the most effective protein stabilizer, we here report TMAO-derived zwitterionic polymers (PTMAO) as a new class of ultralow fouling biomaterials. The nonfouling properties of PTMAO were demonstrated under highly challenging conditions. The mechanism accounting for the extraordinary hydration of PTMAO was elucidated by molecular dynamics simulations. The discovery of PTMAO polymers demonstrates the power of molecular understanding in the design of new biomimetic materials and provides the biomaterials community with another class of nonfouling zwitterionic materials.
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Materiais Biocompatíveis/química , Incrustação Biológica/prevenção & controle , Metilaminas/química , Polímeros/química , Adsorção , Animais , Materiais Biocompatíveis/metabolismo , Materiais Biocompatíveis/farmacologia , Adesão Celular/efeitos dos fármacos , Humanos , Metilaminas/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Simulação de Dinâmica Molecular , Células NIH 3T3 , Polímeros/metabolismo , Polímeros/farmacologia , Albumina Sérica/química , Ressonância de Plasmônio de SuperfícieRESUMO
Nerve agents are a class of organophosphorus compounds (OPs) that blocks communication between nerves and organs. Because of their acute neurotoxicity, it is extremely difficult to rescue the victims after exposure. Numerous efforts have been devoted to search for an effective prophylactic nerve agent bioscavenger to prevent the deleterious effects of these compounds. However, low scavenging efficiency, unfavorable pharmacokinetics, and immunological problems have hampered the development of effective drugs. Here, we report the development and testing of a nanoparticle-based nerve agent bioscavenger (nanoscavenger) that showed long-term protection against OP intoxication in rodents. The nanoscavenger, which catalytically breaks down toxic OP compounds, showed a good pharmacokinetic profile and negligible immune response in a rat model of OP intoxication. In vivo administration of the nanoscavenger before or after OP exposure in animal models demonstrated protective and therapeutic efficacy. In a guinea pig model, a single prophylactic administration of the nanoscavenger effectively prevented lethality after multiple sarin exposures over a 1-week period. Our results suggest that the prophylactic administration of the nanoscavenger might be effective in preventing the toxic effects of OP exposure in humans.
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Nanopartículas/química , Agentes Neurotóxicos/toxicidade , Substâncias Protetoras/farmacologia , Administração Intravenosa , Animais , Feminino , Cobaias , Masculino , Nanopartículas/administração & dosagem , Paraoxon/toxicidade , Substâncias Protetoras/administração & dosagem , Substâncias Protetoras/farmacocinética , Ratos Sprague-Dawley , Sarina/toxicidade , Análise de Sobrevida , Fatores de Tempo , Distribuição TecidualRESUMO
Here, we report a simple yet effective surface-modification approach to imparting hydrophobic surfaces with superhydrophilicity using ultralow fouling/functionalizable carboxybetaine (CB) copolymers via a dip-coating technique. A new series of CB random copolymers with varying amphiphilicities were synthesized and coated on hydrophobic polypropylene (PP) and polystyrene (PS) surfaces. The nonfouling capability of each coating was screened by an enzyme-linked immunosorbent assay (ELISA) and further comprehensively assessed against 100% human serum by a Micro BCA protein assay kit. The random copolymer containing â¼30 mol % CB units showed superhydrophilicity with the highest air contact angle of more than 165° in DI water and the best nonfouling capability against 100% human blood serum. Surfaces of a 96-well plate coated with the optimal CB random copolymer had a significantly better nonfouling capability than those of a commercial 96-well plate with an ultralow attachment surface. The adhesion of mouse embryonic fibroblast cells (NIH3T3) was completely inhibited on surfaces coated with CB random copolymers. Furthermore, the optimal nonfouling CB copolymer surface was functionalized with an antigen via covalent bonding where its specific interactions with its antibody were verified. Thus, this CB random copolymer is capable of imparting both ultralow fouling and functionalizable capabilities to hydrophobic surfaces for blood-contacting devices.
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Resinas Acrílicas/química , Incrustação Biológica/prevenção & controle , Compostos de Amônio Quaternário/química , Resinas Acrílicas/síntese química , Resinas Acrílicas/metabolismo , Adsorção , Animais , Proteínas Sanguíneas/metabolismo , Humanos , Camundongos , Células NIH 3T3 , Polipropilenos/química , Poliestirenos/química , Ligação Proteica , Compostos de Amônio Quaternário/síntese química , Compostos de Amônio Quaternário/metabolismoRESUMO
Much attention has been drawn to targeted nanodrug delivery systems due to their high therapeutic efficacy in cancer treatment. In this work, doxorubicin (DOX) was incorporated into a zwitterionic arginyl-glycyl-aspartic acid (RGD)-conjugated polypeptide by an emulsion solvent evaporation technique with high drug loading content (45%) and high drug loading efficiency (95%). This zwitterionic nanoformulation showed excellent colloidal stability at high dilution and in serum. The pH-induced disintegration and enzyme-induced degradation of the nanoformulation were confirmed by dynamic light scattering and gel permeation chromatography. Efficient internalization of DOX in the cells and high antitumor activity in vitro was observed. Compared with the free drug, this nanoformulation showed higher accumulation in tumor and lower systemic toxicity in vivo. The DOX-loaded zwitterionic RGD-conjugated polypeptide vesicles show potential application for targeted drug delivery in the clinic.
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Antineoplásicos/farmacologia , Doxorrubicina/farmacologia , Portadores de Fármacos/química , Peptídeos Cíclicos/química , Ácido Poliglutâmico/análogos & derivados , Polilisina/análogos & derivados , Linhagem Celular Tumoral , Portadores de Fármacos/toxicidade , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Humanos , Nanopartículas/química , Nanopartículas/toxicidade , Peptídeos Cíclicos/toxicidade , Ácido Poliglutâmico/química , Ácido Poliglutâmico/toxicidade , Polilisina/química , Polilisina/toxicidadeRESUMO
Conjugation with poly(ethylene glycol) (PEG) or PEGylation is a widely used tool to overcome the shortcomings of native proteins, such as poor stability, inadequate pharmacokinetic (PK) profiles, and immunogenicity. However, PEGylation is often accompanied by an unwanted detrimental effect on bioactivity, particularly, resulting from the amphiphilic nature of PEG. This is especially true for PEGylated proteins with large binding targets. Pegasys, a PEGylated interferon alpha-2a (IFN-α2a) bearing a 40 kDa branched PEG, is a typical example that displays only 7% in vitro activity of the unmodified IFN-α2a. In this work, by employing IFN-α2a as a model protein, we demonstrated that a protein conjugated with zwitterionic polymers (or zwitterlation) could significantly mitigate the antiproliferative bioactivity loss in vitro after polymer conjugation. The retained antiproliferative activity of zwitterlated IFN-α2a is 4.4-fold higher than that of the PEGylated IFN-α2a with the same polymer molecular weight, or 3-fold higher than that of the PEGylated IFN-α2a with a similar hydrodynamic size. It is hypothesized that nonspecific interactions between zwitterionic polymers and IFN-α2a/IFN-α2a receptors can be mitigated due to the super-hydrophilic nature of zwitterionic polymers. This, in turn, reduces the 'nonspecific blocking' between IFN-α2a and IFN-α2a receptors. In addition, we demonstrated that zwitterlated IFN-α2a showed a prolonged circulation time and a mitigated accelerated blood clearance after repeated injections in rats.
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The undesirable immune response poses a life-threatening challenge to human health. It not only deteriorates the therapeutic performance of biologic drugs but also contributes to various diseases such as allergies and autoimmune diseases. Inspired by the role of chromatin in the maintenance of natural immune tolerance, here we report a DNA-protein polymeric nanocomplex that can mimic the tolerogenic function of chromatin and induce an immune tolerance to its protein cargos. We first proved that the chromatin-mimetic nanomedicine loaded with keyhole limpet hemocyanin (KLH), a highly immunogenic model protein, could elicit a durable antigen-specific immune tolerance to KLH lasting for at least five weeks in mice. Following the proof-of-concept study, we demonstrated that this nanomedicine could be applied to improve the safety and efficacy of a biologic drug, PEGylated uricase, by attenuating the relevant antibody (Ab) responses. Moreover, we also demonstrated that prophylactic treatments with this nanomedicine could tolerize the immune system with the allergen of ovalbumin (OVA) and thus inhibit the occurrence of airway inflammation in an OVA-induced allergic asthma murine model. Collectively, our work illustrates a nature-inspired concept of immune tolerance induction and establishes a useful tool to specifically suppress unwanted immune responses for therapeutic purposes.
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Asma/imunologia , Cromatina/imunologia , Hemocianinas/imunologia , Tolerância Imunológica/imunologia , Nanomedicina , Animais , Asma/induzido quimicamente , Cromatina/química , Modelos Animais de Doenças , Hemocianinas/química , Masculino , Camundongos , Camundongos Endogâmicos C57BL , OvalbuminaRESUMO
Poly(ethylene glycol) (PEG) conjugation has been the gold standard to ameliorate the pharmacokinetic (PK) and immunological profiles of proteins. PEG polymer does become immunogenic once attached to proteins, evoking PEG-specific antibody (Ab) responses. The anti-PEG Abs could cause PEGylated biologic treatments to fail and even result in lethal adverse reactions. Thus the zwitterionic poly(carboxybetaine) (PCB) has been introduced as a PEG substitute for protein modification. Addressed herein is anti-polymer Ab induction by conjugating PEG and PCB polymers to a series of carrier proteins with escalating immunogenicity. Results indicate that titers of PEG-specific Abs were quantitatively correlated to the immunogenicity of carrier proteins, whereas the generation of PCB-specific Abs was minimal and insensitive to increased protein immunogenicity. This work provides insight into the immunological properties of PEG and PCB and has far-reaching implications for the development of polymer-protein conjugates.