ACS Pharmacology and Translational Science最新文献

Background: Parkinson's disease (PD) is intricately linked to gastrointestinal inflammation and the presence of neurotoxins in the gut, integrating α-syn pathologic alterations and subsequent neurodegeneration into the brain. Objectives: This study aimed to explore the enduring impact of dextran sodium sulfate (DSS)-mediated colitis on the vulnerability of central dopaminergic neurons to subsequent rotenone exposure. Methods: To induce chronic colitis, 10-month-old C57BL/6 mice were pre-exposed to 3 cycles of 1 week of 1% (w/v) DSS administration in drinking water followed by 2 weeks of regular drinking water. After colitis induction, animals received a low dose of intragastric rotenone for the next 8 weeks, followed by testing for Parkinsonian behavior and GI phenotypes of inflammation. At the end of the 17th week, colon, brain stem, and midbrain tissue were isolated and analyzed for α-syn, inflammatory markers, and dopaminergic neuronal loss. Gut microbial composition was assessed by 16S rRNA sequencing analysis. Results: We found that chronic rotenone administration in the presence of preexisting colitis led to a further increase in colonic pro-inflammatory mediator expressions, α-syn expression, and reduced colonic tight junction protein expressions. We also found early impairment of GI functions and worsened grip strength in rotenone-exposed colitic mice. Furthermore, α-syn pathology specific to the colitic mice exposed to rotenone showed dopaminergic neurons degeneration and astroglial activation in substantia nigra and striatum, including regions of the brain stem, i.e., dorsal motor of the vagus and locus coeruleus. Finally, the result of 16S rRNA gene sequencing analysis indicated that colitic mice, after being exposed to rotenone, exhibited a discernible trend in their microbiota composition (Catenibacterium, Turicibactor, and clostridium sensue stricto 1), linking it to the development of PD. Conclusions: These findings indicate that prolonged low-dose rotenone exposure, combined with an early inflammatory intestinal milieu, provides a preconditioning effect on α-syn pathology and exerts neurodegeneration in the intragastric rotenone PD mouse model.

mRNA-based therapies can overcome several challenges faced by traditional therapies in treating a variety of diseases by selectively modulating genes and proteins without genomic integration. However, due to mRNA’s poor stability and inherent limitations, nanoparticle (NP) platforms have been developed to deliver functional mRNA into cells. In cancer treatment, mRNA technology has multiple applications, such as restoration of tumor suppressors and activating antitumor immunity. Most of these applications have been evaluated using simple cell-line-based tumor models, which failed to represent the complexity, heterogeneity, and 3D architecture of patient tumors. This discrepancy has led to inconsistencies and failures in clinical translation. Compared to cell line models, patient-derived xenograft (PDX) models more accurately represent patient tumors and are better suitable for modeling. Therefore, for the first time, this study employed two different TNBC PDX tumors to examine the effects of the mRNA-NPs. mRNA-NPs are developed using EGFP-mRNA as a model and studied in TNBC cell lines, ex vivo TNBC PDX organotypic slice cultures, and in vivo TNBC PDX tumors. Our findings show that NPs can effectively accumulate in tumors after intravenous administration, protecting and delivering mRNA to PDX tumors with different genetic and chemosensitivity backgrounds. These studies offer more clinically relevant modeling systems for mRNA nanotherapies in cancer applications.

While identification of epigenetic changes in individuals with psychiatric dysfunctions such as post-traumatic stress disorder (PTSD) is paramount to genomic research, there is no rapid and simplified way to detect an epigenetic marker such as DNA methylation in genes. Here, we introduce a faster, simpler method to detect methylation in the form of 5-methylcytosine (5mC, termed as PTSD-associated base) in known C_pG sites using nanoenhanced surface plasmon resonance imaging-based epigenetic assay (EpiNanoSPRi). This assay platform simultaneously detects a panel of single, site-specific PTSD bases in target genes or regions using an anti-5mC antibody and a universal nanoenhancer on a gold-coated sensing chip. Not only can EpiNanoSPRi identify 5mC at the single-base level, but it also can quantify the extent of DNA methylation. Our method is superior and more practical to bisulfite-based DNA sequencing techniques as it will significantly reduce DNA methylation identification from 4 days (e.g., DNA Sequencing) to 9 h without massive analysis workflow. This platform can potentially be applied to diagnose other psychiatric disorders such as Alzheimer’s, Parkinson’s, dementia, schizophrenia, and Huntington’s diseases.

Background: Parkinson’s disease (PD) is intricately linked to gastrointestinal inflammation and the presence of neurotoxins in the gut, integrating α-syn pathologic alterations and subsequent neurodegeneration into the brain. Objectives: This study aimed to explore the enduring impact of dextran sodium sulfate (DSS)-mediated colitis on the vulnerability of central dopaminergic neurons to subsequent rotenone exposure. Methods: To induce chronic colitis, 10-month-old C57BL/6 mice were pre-exposed to 3 cycles of 1 week of 1% (w/v) DSS administration in drinking water followed by 2 weeks of regular drinking water. After colitis induction, animals received a low dose of intragastric rotenone for the next 8 weeks, followed by testing for Parkinsonian behavior and GI phenotypes of inflammation. At the end of the 17th week, colon, brain stem, and midbrain tissue were isolated and analyzed for α-syn, inflammatory markers, and dopaminergic neuronal loss. Gut microbial composition was assessed by 16S rRNA sequencing analysis. Results: We found that chronic rotenone administration in the presence of preexisting colitis led to a further increase in colonic pro-inflammatory mediator expressions, α-syn expression, and reduced colonic tight junction protein expressions. We also found early impairment of GI functions and worsened grip strength in rotenone-exposed colitic mice. Furthermore, α-syn pathology specific to the colitic mice exposed to rotenone showed dopaminergic neurons degeneration and astroglial activation in substantia nigra and striatum, including regions of the brain stem, i.e., dorsal motor of the vagus and locus coeruleus. Finally, the result of 16S rRNA gene sequencing analysis indicated that colitic mice, after being exposed to rotenone, exhibited a discernible trend in their microbiota composition (Catenibacterium, Turicibactor, and clostridium sensue stricto 1), linking it to the development of PD. Conclusions: These findings indicate that prolonged low-dose rotenone exposure, combined with an early inflammatory intestinal milieu, provides a preconditioning effect on α-syn pathology and exerts neurodegeneration in the intragastric rotenone PD mouse model.

mRNA-based therapies can overcome several challenges faced by traditional therapies in treating a variety of diseases by selectively modulating genes and proteins without genomic integration. However, due to mRNA's poor stability and inherent limitations, nanoparticle (NP) platforms have been developed to deliver functional mRNA into cells. In cancer treatment, mRNA technology has multiple applications, such as restoration of tumor suppressors and activating antitumor immunity. Most of these applications have been evaluated using simple cell-line-based tumor models, which failed to represent the complexity, heterogeneity, and 3D architecture of patient tumors. This discrepancy has led to inconsistencies and failures in clinical translation. Compared to cell line models, patient-derived xenograft (PDX) models more accurately represent patient tumors and are better suitable for modeling. Therefore, for the first time, this study employed two different TNBC PDX tumors to examine the effects of the mRNA-NPs. mRNA-NPs are developed using EGFP-mRNA as a model and studied in TNBC cell lines, ex vivo TNBC PDX organotypic slice cultures, and in vivo TNBC PDX tumors. Our findings show that NPs can effectively accumulate in tumors after intravenous administration, protecting and delivering mRNA to PDX tumors with different genetic and chemosensitivity backgrounds. These studies offer more clinically relevant modeling systems for mRNA nanotherapies in cancer applications.

While identification of epigenetic changes in individuals with psychiatric dysfunctions such as post-traumatic stress disorder (PTSD) is paramount to genomic research, there is no rapid and simplified way to detect an epigenetic marker such as DNA methylation in genes. Here, we introduce a faster, simpler method to detect methylation in the form of 5-methylcytosine (5mC, termed as PTSD-associated base) in known C_pG sites using nanoenhanced surface plasmon resonance imaging-based epigenetic assay (EpiNanoSPRi). This assay platform simultaneously detects a panel of single, site-specific PTSD bases in target genes or regions using an anti-5mC antibody and a universal nanoenhancer on a gold-coated sensing chip. Not only can EpiNanoSPRi identify 5mC at the single-base level, but it also can quantify the extent of DNA methylation. Our method is superior and more practical to bisulfite-based DNA sequencing techniques as it will significantly reduce DNA methylation identification from 4 days (e.g., DNA Sequencing) to 9 h without massive analysis workflow. This platform can potentially be applied to diagnose other psychiatric disorders such as Alzheimer's, Parkinson's, dementia, schizophrenia, and Huntington's diseases.

Skin cancer represents a major health concern due to its rising incidence and limited treatment options. Current treatments (surgery, chemotherapy, radiotherapy, immunotherapy, and targeted therapy) often entail high costs, patient inconvenience, significant adverse effects, and limited therapeutic efficacy. The search for novel treatment options is also marked by the high capital investment and extensive development involved in the drug discovery process. In response to these challenges, repurposing existing drugs for topical application and optimizing their delivery through nanotechnology could be the answer. This innovative strategy aims to combine the advantages of the known pharmacological background of commonly used drugs to expedite therapeutic development, with nanosystem-based formulations, which among other advantages allow for improved skin permeation and retention and overall higher therapeutic efficacy and safety. The present review provides a critical analysis of repurposed drugs such as doxycycline, itraconazole, niclosamide, simvastatin, leflunomide, metformin, and celecoxib, formulated into different nanosystems, namely, nanoemulsions and nanoemulgels, nanodispersions, solid lipid nanoparticles, nanostructured lipid carriers, polymeric nanoparticles, hybrid lipid–polymer nanoparticles, hybrid electrospun nanofibrous scaffolds, liposomes and liposomal gels, ethosomes and ethosomal gels, and aspasomes, for improved outcomes in the battle against skin cancer. Enhanced antitumor effects on melanoma and nonmelanoma research models are highlighted, with some nanoparticles even showing intrinsic anticancer properties, leading to synergistic effects. The explored research findings highly evidence the potential of these approaches to complement the currently available therapeutic strategies in the hope that these treatments might one day reach the pharmaceutical market.

Fusion proteins constitute a class of engineered therapeutics and have emerged as promising candidates for disease treatment. However, the structural complexity and heterogeneity of fusion proteins make their characterization extremely challenging, and thus, an innovative and comprehensive analytical toolbox is needed. Here, for the first time, we demonstrate a novel and robust workflow to evaluate charge variants for a highly glycosylated fusion protein with heavy sialylation using imaged capillary isoelectric focusing (icIEF). In the development of the icIEF method, key factors that were systematically investigated include the desialylation level, the stability of the desialylated molecule, incubation time and temperature of desialylation, protein concentrations, urea and l-arginine effects on the tertiary structure, and instrumental comparability. Multivariate and correlation analyses were subsequently applied to confirm the impacts of the parameters evaluated. Furthermore, a microfluidic chip-based icIEF system coupled with ultraviolet detection and mass spectrometry (icIEF-UV/MS) was utilized to identify critical post-translational modifications and ameliorate the understanding of charge variants. Our study demonstrates that this workflow enables a mechanistic understanding of charge variants for heavily sialylated therapeutics.

Skin cancer represents a major health concern due to its rising incidence and limited treatment options. Current treatments (surgery, chemotherapy, radiotherapy, immunotherapy, and targeted therapy) often entail high costs, patient inconvenience, significant adverse effects, and limited therapeutic efficacy. The search for novel treatment options is also marked by the high capital investment and extensive development involved in the drug discovery process. In response to these challenges, repurposing existing drugs for topical application and optimizing their delivery through nanotechnology could be the answer. This innovative strategy aims to combine the advantages of the known pharmacological background of commonly used drugs to expedite therapeutic development, with nanosystem-based formulations, which among other advantages allow for improved skin permeation and retention and overall higher therapeutic efficacy and safety. The present review provides a critical analysis of repurposed drugs such as doxycycline, itraconazole, niclosamide, simvastatin, leflunomide, metformin, and celecoxib, formulated into different nanosystems, namely, nanoemulsions and nanoemulgels, nanodispersions, solid lipid nanoparticles, nanostructured lipid carriers, polymeric nanoparticles, hybrid lipid-polymer nanoparticles, hybrid electrospun nanofibrous scaffolds, liposomes and liposomal gels, ethosomes and ethosomal gels, and aspasomes, for improved outcomes in the battle against skin cancer. Enhanced antitumor effects on melanoma and nonmelanoma research models are highlighted, with some nanoparticles even showing intrinsic anticancer properties, leading to synergistic effects. The explored research findings highly evidence the potential of these approaches to complement the currently available therapeutic strategies in the hope that these treatments might one day reach the pharmaceutical market.

Fusion proteins constitute a class of engineered therapeutics and have emerged as promising candidates for disease treatment. However, the structural complexity and heterogeneity of fusion proteins make their characterization extremely challenging, and thus, an innovative and comprehensive analytical toolbox is needed. Here, for the first time, we demonstrate a novel and robust workflow to evaluate charge variants for a highly glycosylated fusion protein with heavy sialylation using imaged capillary isoelectric focusing (icIEF). In the development of the icIEF method, key factors that were systematically investigated include the desialylation level, the stability of the desialylated molecule, incubation time and temperature of desialylation, protein concentrations, urea and l-arginine effects on the tertiary structure, and instrumental comparability. Multivariate and correlation analyses were subsequently applied to confirm the impacts of the parameters evaluated. Furthermore, a microfluidic chip-based icIEF system coupled with ultraviolet detection and mass spectrometry (icIEF-UV/MS) was utilized to identify critical post-translational modifications and ameliorate the understanding of charge variants. Our study demonstrates that this workflow enables a mechanistic understanding of charge variants for heavily sialylated therapeutics.