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Personalized cancer vaccines targeting specific neoantigens have been envisioned as one of the most promising approaches in cancer immunotherapy. However, the physicochemical variability of the identified neoantigens limits their efficacy as well as vaccine manufacturing in a uniform format. Herein, we developed a uniform nanovaccine platform based on poly(2-oxazoline)s (POx) to chemically conjugate neoantigen peptides, regardless of their physicochemical properties. This vaccine system could self-assemble into nanoparticles with uniform size (around 50 nm) and improve antigen accumulation as well as infiltration in the lymph node to increase antigen presentation. In vivo vaccination using this system conjugated with three predicted peptide neoantigen peptides from the MC38 tumor cell line induced 100% robust CD8+ T cell responses and superior tumor clearance compared to free peptides. This POx-based vaccine carrier represents a generalizable approach to increase the availability and efficacy of screened neoantigen peptides for a personalized cancer vaccine.
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Antígenos de Neoplasias , Vacunas contra el Cáncer , Nanopartículas , Péptidos , Vacunas contra el Cáncer/inmunología , Vacunas contra el Cáncer/administración & dosificación , Vacunas contra el Cáncer/química , Péptidos/química , Péptidos/inmunología , Animales , Antígenos de Neoplasias/inmunología , Antígenos de Neoplasias/química , Ratones , Nanopartículas/química , Humanos , Línea Celular Tumoral , Linfocitos T CD8-positivos/inmunología , Oxazoles/química , Polímeros/química , Inmunoterapia/métodos , NanovacunasRESUMEN
Overweight, with an increasing prevalence worldwide, significantly impairs the clinical outcomes following in vitro fertilization (IVF). Hyperglycemia, hyperlipidemia, and metabolic disorders are always accompanied by the majority of overweight patients. The association between granulosa cell function and metabolic alterations in follicular fluid including lipids, proteins, and growth factors has been extensively documented. However, the effects of higher glucose level on ovarian granulosa cells (GCs), remain largely unknown. In this study, we identified that overweight women had elevated follicular glucose level which profoundly activated NLRP3 inflammasome and pyroptosis. An in vitro correlation between follicular high glucose, NLRP3 inflammasome and pyroptosis was also established. More importantly, in granulosa cells of overweight patients, the activation of the NLRP3 inflammasome and pyroptosis induced by high glucose was involved in the dysregulation of estradiol synthesis. Our study may provide new options to interpretate and improve IVF outcomes in overweight women.
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Inflamasomas , Proteína con Dominio Pirina 3 de la Familia NLR , Humanos , Femenino , Inflamasomas/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Glucosa/farmacología , Piroptosis , Sobrepeso , Células de la Granulosa/metabolismoRESUMEN
Robotic Mobile Fulfillment Systems (RMFSs) face challenges in handling large-scale orders and navigating complex environments, frequently encountering a series of intricate decision-making problems, such as order allocation, shelf selection, and robot scheduling. To address these challenges, this paper integrates Deep Reinforcement Learning (DRL) technology into an RMFS, to meet the needs of efficient order processing and system stability. This study focuses on three key stages of RMFSs: order allocation and sorting, shelf selection, and coordinated robot scheduling. For each stage, mathematical models are established and the corresponding solutions are proposed. Unlike traditional methods, DRL technology is introduced to solve these problems, utilizing a Genetic Algorithm and Ant Colony Optimization to handle decision making related to large-scale orders. Through simulation experiments, performance indicators-such as shelf access frequency and the total processing time of the RMFS-are evaluated. The experimental results demonstrate that, compared to traditional methods, our algorithms excel in handling large-scale orders, showcasing exceptional superiority, capable of completing approximately 110 tasks within an hour. Future research should focus on integrated decision-making modeling for each stage of RMFSs and designing efficient heuristic algorithms for large-scale problems, to further enhance system performance and efficiency.
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Virus-like nanoparticle vaccines can efficiently activate the humoral immune response by cross-linking B cell receptors with their surface multivalent antigen arrays. This structurally dependent mechanism makes it crucial to regulate and optimize structural parameters to enhance the efficacy of nanoparticle vaccines. In this study, we prepared nanoparticle vaccines with different aspect ratios by chemically modifying antigen proteins onto the surfaces of poly(amino acid) nanoparticles of various shapes (spherical, ellipsoidal, and rod-like). This allowed us to investigate the impact of structural anisotropy on the humoral immune activation efficacy of nanoparticle vaccines. Furthermore, the end-group molecules of poly(amino acid) materials possess aggregation-induced emission (AIE) properties, which facilitate monitoring the dynamics of nano-assemblies within the body. Results showed that rod-like nanoparticle vaccines (RLNVax) with a higher aspect ratio (AR = 5) exhibited greater lymph node draining efficiency and could elicit more effective B cell activation compared to conventional isotropic spherical nanoparticle vaccines. In a murine subcutaneous immunization model using ovalbumin (OVA) as a model antigen, RLNVax elicited antigen-specific antibody titers that were about 64 times and 4.6 times higher than those induced by free antigen proteins and spherical nanoparticle vaccines, respectively. Additionally, when combined with an aluminum adjuvant, antibody titers elicited by RLNVax were further enhanced by 4-fold. These findings indicate that the anisotropic rod-like structure is advantageous for improving the humoral immune activation efficacy of nanoparticle vaccines, providing significant insights for the design and optimization of next-generation nanoparticle vaccines.
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Nanoparticles have been regarded as a promising vaccine adjuvant due to their innate immune potentiation and enhanced antigen transport. However, the inefficient infiltration into the lymph node (LN) paracortex of nanoparticles caused by subcapsular sinus (SCS) obstruction is the main challenge in further improvement of nanovaccine immune efficacy. Herein, we propose to overcome paracortex penetration by using nanovaccine to spontaneously and continuously release antigens after retention in the SCS. In detail, we utilized a spontaneous retro-Diels-Alder (r-D-A) reaction linker to connect poly{(2-methyl-2-oxazoline)80-co-[(2-butyl-2-oxazoline)15-r-(2-thioethyl-2-oxazoline)8]} (PMBOxSH) and peptides for the peptide nanovaccine construction. The r-D-A reaction linker can spontaneously break over time, allowing the nanovaccine to release free antigens and adjuvants upon reaching the LN, thereby facilitating the entry of released antigens and adjuvants into the interior of the LNs. We showed that the efficacy of the peptide nanovaccine constructed using this dynamic linker could be significantly improved, thus greatly enhancing the tumor inhibition efficacy in the B16-OVA model. This dynamic-covalent-chemistry-based vaccine strategy may inspire designing more efficient therapeutic vaccines, especially those that require eliciting high-amount T cell responses.
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Inmunidad Celular , Ganglios Linfáticos , Nanopartículas , Péptidos , Animales , Ratones , Ganglios Linfáticos/efectos de los fármacos , Ganglios Linfáticos/inmunología , Nanopartículas/química , Péptidos/química , Péptidos/farmacología , Inmunidad Celular/efectos de los fármacos , Ratones Endogámicos C57BL , Vacunas contra el Cáncer/química , Vacunas contra el Cáncer/inmunología , Vacunas contra el Cáncer/administración & dosificación , Reacción de Cicloadición , Femenino , Tamaño de la Partícula , NanovacunasRESUMEN
Virus-like particle (VLP) vaccines had shown great potential during the COVID-19 pandemic, and was thought to be the next generation of antiviral vaccine technology due to viromimetic structures. However, the time-consuming and complicated processes in establishing a current recombinant-protein-based VLP vaccine has limited its quick launch to the out-bursting pandemic. To simplify and optimize VLP vaccine design, we herein report a kind of viromimetic polymer nanoparticle vaccine (VPNVax), with subunit receptor-binding domain (RBD) proteins conjugated to the surface of polyethylene glycol-b-polylactic acid (PEG-b-PLA) nanoparticles for vaccination against SARS-CoV-2. The preparation of VPNVax based on synthetic polymer particle and chemical post-conjugation makes it possible to rapidly replace the antigens and construct matched vaccines at the emergence of different viruses. Using this modular preparation system, we identified that VPNVax with surface protein coverage of 20%-25% had the best immunostimulatory activity, which could keep high levels of specific antibody titers over 5 months and induce virus neutralizing activity when combined with an aluminum adjuvant. Moreover, the polymer nano-vectors could be armed with more immune-adjuvant functions by loading immunostimulant agents or chemical chirality design. This VPNVax platform provides a novel kind of rapidly producing and efficient vaccine against different variants of SARS-CoV-2 as well as other viral pandemics.
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BACKGROUND: Oral gonadotropin-releasing hormone (GnRH) antagonists are promising agents in the treatment of endometriosis-related pain. Here we assessed the safety, pharmacokinetics (PK), and pharmacodynamics (PD) of SHR7280, an oral non-peptide GnRH antagonist in premenopausal women with endometriosis. METHODS: In the Phase 1 part of the randomized, double-blinded, placebo-controlled, dose-ascending, Phase 1/2 trial, premenopausal women with endometriosis were randomized (4:1) to receive SHR7280 or placebo treatment for 21 consecutive days. The treatment dose started from 200 mg QD, and then increased to 300 mg QD and 200 mg BID. Safety, PK, and PD parameters were assessed. RESULTS: In total, 30 patients received assigned treatment, 24 with SHR7280 and 6 with placebo. SHR7280 was well tolerated. Adverse events (AEs) were reported in 19 (79.2%, 19/24) patients in the SHR7280 group and 5 (83.3%, 5/6) patients in the placebo group. Most AEs were mild and no severe AEs occurred. SHR7280 showed a rapid absorption, with a time to maximum plasma concentration (Tmax) of 1.0 h, 1.0 h, and 0.8 h for the 200 mg QD, 300 mg QD, and 200 mg BID regimens, respectively. Plasma concentration of SHR7280 was dose dependent. The mean half-life (t1/2) at steady state was 6.9 h, 7.4 h, and 2.8 h, respectively, and little or no accumulation was observed. Pharmacodynamic analysis showed that SHR7280 could effectively suppress estradiol and luteinizing hormone concentrations and prevent progesterone increase in a dose-dependent manner. SHR7280 at doses of 300 mg QD and 200 mg BID could suppress estradiol levels within the desired therapeutic window of 20-50 pg/mL throughout the treatment period. CONCLUSIONS: SHR7280 showed favorable safety, PK, and PD profiles in the doses of 200 mg QD, 300 mg QD, and 200 mg BID. The results of this study provide evidence to support the further development of SHR7280 as a GnRH antagonist for the treatment of endometriosis-related pain in the subsequent Phase 2 trial. TRIAL REGISTRY: Trial registration number: Clinicaltrials.gov, identifier: NCT04417972. Trial registration date: 5 June 2020.
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Endometriosis , Humanos , Femenino , Endometriosis/tratamiento farmacológico , Antagonistas de Hormonas/efectos adversos , Estradiol/uso terapéutico , Dolor , Método Doble Ciego , Hormona Liberadora de Gonadotropina , Relación Dosis-Respuesta a DrogaRESUMEN
The residues of fipronil and its metabolites, namely fipronil-desulfinyl, fipronil-sulfone, and fipronil-sulfide, have attracted increasing attention since the European egg contamination incident. In this study, by optimizing the pretreatment method and chromatographic conditions, a simple extraction method coupled with high performance liquid chromatography-tandem mass spectrometry was developed for the simultaneous determination of fipronil and its metabolites, including fipronil-desulfinyl, fipronil-sulfone, and fipronil-sulfide, in livestock and poultry liver. The optimal pretreatment method for fipronil and its metabolites was determined by comparing the recoveries obtained with different extraction solvents (methanol, acetonitrile, acetone, and ethyl acetate), and by purification with N-propyl ethylenediamine (PSA) and octadecylsilane (C18). The samples were extracted with 10 mL acetonitrile, then purified with 150 mg PSA and 100 mg C18, following which the extracted solutions were directly injected for analysis. Separation was performed on an Agilent ZORBAX SB-C18 column (150 mm×2.1 mm, 3.5 µm) with gradient elution using acetonitrile and water as the mobile phases. The target compounds were detected by electrospray ionization (ESI) in the negative mode under multiple reaction monitoring, and quantified by the external standard method with matrix-matched standard correction curves. The results indicated that the linear ranges for the four compounds ranged from 0.1 µg/L to 10.0 µg/L with correlation coefficients (r2) higher than 0.995. The limit of detection was 0.2 µg/kg and limit of quantitation was 0.5 µg/kg. The average recoveries were between 81.1% and 99.8% at three spiked levels of 0.5, 1.0, and 10 µg/kg, with relative standard deviations (RSDs) of 6.1%-11.7%. The matrix effect experiment showed that fipronil and its metabolites had matrix inhibition effects. Matrix-matched standard curve correction was performed to eliminate matrix inhibition effects. The proposed method was used for the analysis of 99 liver samples, where fipronil-sulfone was detected in four samples with values of 1.25-2.82 µg/kg. The method is simple, sensitive, and accurate, and is suitable for the determination of fipronil and its metabolites in livestock and poultry liver.
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Ganado , Espectrometría de Masas en Tándem , Animales , Cromatografía Líquida de Alta Presión , Hígado , Aves de Corral , Pirazoles , Extracción en Fase SólidaRESUMEN
Decidualization is driven by differentiation of human endometrial stromal cells (ESCs), and is a prerequisite for successful implantation and establishment of pregnancy. The critical role of impaired decidualization in women suffered recurrent implantation failure (RIF) has been established, while the underlying mechanism is poorly understood. In the present study, we verified the essential role of Sirtuin1 (SIRT1) in regulating differentiation and maintaining reactive oxygen species (ROS) homeostasis of human ESCs during decidualization. The abundance of SIRT1 was decreased in RIF patients both in the endometria during window of implantation phase and in the decidualized ESCs. Downregulation of SIRT1 disrupted the intracellular ROS homeostasis during decidualization of ESC, manifested as the accumulation of intracellular ROS level and the reduction of antioxidant stress molecules. Elimination of ROS with N-acetyl-L-cysteine (NAC) could rescued the decidualization inhibition caused by SIRT1 knockdown. Further, we explored the insufficient expression of SIRT1 in ESC affected the deacetylation of forkhead box O1 (FOXO1), and thus inhibited the transcriptional activity of FOXO1. This could account for the dysregulation of intracellular ROS homeostasis during decidualization and decreased expression of decidual markers. Collectively, our findings provided insight into the role of down-regulated SIRT1 in the poor decidual response of ESCs in RIF patients.
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BACKGROUND: Polycystic ovary syndrome (PCOS) is the most common metabolic and endocrine disorder among reproductive-age women, and the leading cause of anovulatory infertility. 11ß-hydroxysteroid dehydrogenases-1 (11ß-HSD1) catalysing the conversion of inactive cortisone to active cortisol plays a crucial role in various metabolic diseases. However, whether 11ß-HSD1 is associated with the pathogenesis of PCOS and whether 11ß-HSD1 can be a treating target of PCOS remain unknown. METHODS: This study was first designed to explore the role of 11ß-HSD1 in PCOS development and the effect of selective 11ß-HSD1 inhibitor administration on PCOS treatment. Follicular fluid and granulosa cells (GCs) were collected from 32 non-PCOS patients and 37 patients with PCOS to measure cortisol and 11ß-HSDs levels. Female Sprague-Dawley rats (3-week-old) were injected with dehydroepiandrosterone (DHEA) to induce PCOS and their ovaries were collected to measure the abundance of corticosterone (CORT) and 11ß-HSDs. To determine the role of 11ß-HSD1 in PCOS development, we overexpressed 11ß-HSD1 in the ovaries of female rats (5-week-old) or knocked down the expression of 11ß-HSD1 in the ovaries from PCOS rats via lentivirus injection. After lentivirus infection, the body weights, ovarian weights, estrous cycles, reproductive hormones and morphology of the ovary were analysed in rats from different experimental groups. Then to figure out the translational potential of the selective 11ß-HSD1 inhibitor in treating PCOS, PCOS rats were treated with BVT.2733, a selective 11ß-HSD1 inhibitor and a cluster of PCOS-like traits were analysed, including insulin sensitivity, ovulatory function and fertility of rats from the Control, PCOS and PCOS+BVT groups. Rat ovarian explants and human GCs were used to explore the effect of CORT or cortisol on ovarian extracellular matrix remodelling. RESULTS: The elevated expression of 11ß-HSD1 contributed to the increased cortisol and corticosterone (CORT) concentrations observed in the ovaries of PCOS patients and PCOS rats respectively. Our results showed that ovarian overexpression of 11ß-HSD1 induced a cluster of PCOS phenotypes in rats including irregular estrous cycles, reproductive hormone dysfunction and polycystic ovaries. While knockdown of ovarian 11ß-HSD1 of PCOS rats reversed these PCOS-like changes. Additionally, the selective 11ß-HSD1 inhibitor BVT.2733 alleviated PCOS symptoms such as insulin resistance (IR), irregular estrous cycles, reproductive hormone dysfunction, polycystic ovaries, ovulatory dysfunction and subfertility. Moreover, we showed that cortisol target ovarian insulin signalling pathway and ovarian extracellular matrix (ECM) remodelling in vivo, in ovarian explants and in GCs. CONCLUSION: Elevated 11ß-HSD1 abundance in ovarian is involved in the pathogenesis of PCOS by impairing insulin signalling pathway and ECM remodelling. Selective inhibition of 11ß-HSD1 ameliorates a cluster of PCOS phenotypes. Our study demonstrates the selective 11ß-HSD1 inhibitor as a novel and promising strategy for the treatment of PCOS.
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11-beta-Hidroxiesteroide Deshidrogenasa de Tipo 1/antagonistas & inhibidores , 11-beta-Hidroxiesteroide Deshidrogenasa de Tipo 1/fisiología , Piperazinas/uso terapéutico , Síndrome del Ovario Poliquístico/tratamiento farmacológico , Sulfonamidas/uso terapéutico , Tiazoles/uso terapéutico , 11-beta-Hidroxiesteroide Deshidrogenasa de Tipo 1/metabolismo , Animales , Células Cultivadas , Modelos Animales de Enfermedad , Evaluación Preclínica de Medicamentos , Inhibidores Enzimáticos/farmacología , Inhibidores Enzimáticos/uso terapéutico , Femenino , Humanos , Infertilidad Femenina/tratamiento farmacológico , Infertilidad Femenina/metabolismo , Infertilidad Femenina/patología , Resistencia a la Insulina/fisiología , Ovario/enzimología , Ovario/metabolismo , Piperazinas/farmacología , Síndrome del Ovario Poliquístico/etiología , Síndrome del Ovario Poliquístico/metabolismo , Síndrome del Ovario Poliquístico/patología , Ratas , Ratas Sprague-Dawley , Sulfonamidas/farmacología , Tiazoles/farmacologíaRESUMEN
Polycystic ovary syndrome (PCOS) is one of the most common endocrine disorders in reproductive-age women. Reduced progesterone levels are associated with luteal phase deficiency in women with PCOS. The levels of C-X-C motif chemokine ligand-14 (CXCL14) were previously reported to be decreased in human-luteinized granulosa (hGL) cells derived from PCOS patients. However, the function of CXCL14 in hGL cells and whether CXCL14 affects the synthesis of progesterone in hGL cells remain unclear. In the present study, the levels of CXCL14 were reduced in follicular fluid and hGL cells in PCOS patients, accompanied by decreased progesterone levels in follicular fluid and decreased steroidogenic acute regulatory (STAR) expression in hGL cells. CXCL14 administration partially reversed the low progesterone production and STAR expression in hGL cells obtained from PCOS patients. In primary hGL cells, CXCL14 upregulated STAR expression and progesterone production. CXCL14 activated the phosphorylation of cyclic adenosine monophosphate response element-binding protein (CREB) and CREB inhibitor attenuated the modulation of StAR expression by CXCL14. P38 and Jun N-terminal kinase (JNK) pathways were also activated by CXCL14 and inhibition of p38 and JNK attenuated the increase of phosphorylation of CREB, STAR expression and progesterone production caused by CXCL14. Our findings revealed the novel role of CXCL14 in upregulation of STAR expression and progesterone synthesis through CREB phosphorylation via activation of p38 and JNK pathways in hGL cells. This is likely contributing to the dysfunction in steroidogenesis in granulosa cells from PCOS patients.
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Quimiocinas CXC/farmacología , Células de la Granulosa/efectos de los fármacos , Células de la Granulosa/metabolismo , Fosfoproteínas/metabolismo , Progesterona/biosíntesis , Adulto , Antracenos/farmacología , Células Cultivadas , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/genética , Enzima de Desdoblamiento de la Cadena Lateral del Colesterol/metabolismo , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/genética , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/metabolismo , Inhibidores Enzimáticos/farmacología , Femenino , Flavonoides/farmacología , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/fisiología , Humanos , Imidazoles/farmacología , Fosfoproteínas/genética , Síndrome del Ovario Poliquístico , Progesterona Reductasa/genética , Progesterona Reductasa/metabolismo , Piridinas/farmacologíaRESUMEN
A high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method was developed for the simultaneous determination of 11 mycotoxins in feeds. The samples were extracted with acetonitrile, then cleaned up by multifunctional purification column filler and PRIME HLB solid phase extraction column. The 11 mycotoxins were separated on an Agilent Zorbax SB-C18 column (150 mm×2.1 mm, 3.5 µ m) with gradient elution program, and methanol-5 mmol/L ammonium acetate solution containing 0.1%(v/v) formic acid were used as mobile phases. The target compounds were detected under electrospray ionization (ESI) both in positive and negative modes with multiple reaction monitoring mode, and quantified by internal standard method. The results indicated that the 11 mycotoxins showed good linear relationships in their respective linear ranges, and the correlation coefficients were greater than 0.99. The limits of quantification (LOQs) were between 2.0 and 50.0 µ g/kg. The average recoveries were between 79.3% and 101.6% at three spiked levels (1, 2 and 5 times LOQs) with relative standard deviations (RSDs) of 5.9%-13.2% (n=5). The method is simple, rapid, sensitive, and can be used for the analysis of the 11 mycotoxins in feeds.