Disruption of myelin, the fatty sheath-insulating nerve fibers in the white matter, blocks or slows the rapid transmission of electrical signals along nerve cells and contributes to several neurodegenerative diseases such as multiple sclerosis. Traditionally, research has focused on neuronal dysfunction as the primary factor, including autoimmunity, infections, inflammation, and genetic disorders causing demyelination. However, recent insights emphasize the critical role of pericytes, non-neuronal cells that regulate blood flow and maintain the health of blood vessels within white matter. This Perspective explores the principal mechanisms through which pericyte dysfunction contributes to damage and demyelination, including impaired communication with neurons (neurovascular uncoupling), excessive formation of scar tissue (fibrosis), and the infiltration of detrimental substances from the bloodstream. Understanding these mechanisms of pericyte-driven demyelination may lead to the creation of new therapeutic strategies for tackling a range of neurodegenerative conditions.
{"title":"The Hidden Hand in White Matter: Pericytes and the Puzzle of Demyelination","authors":"Siddharth Raj, Priyabrata Sarangi, Divya Goyal, Hemant Kumar","doi":"10.1021/acsptsci.4c00192","DOIUrl":"https://doi.org/10.1021/acsptsci.4c00192","url":null,"abstract":"Disruption of myelin, the fatty sheath-insulating nerve fibers in the white matter, blocks or slows the rapid transmission of electrical signals along nerve cells and contributes to several neurodegenerative diseases such as multiple sclerosis. Traditionally, research has focused on neuronal dysfunction as the primary factor, including autoimmunity, infections, inflammation, and genetic disorders causing demyelination. However, recent insights emphasize the critical role of pericytes, non-neuronal cells that regulate blood flow and maintain the health of blood vessels within white matter. This Perspective explores the principal mechanisms through which pericyte dysfunction contributes to damage and demyelination, including impaired communication with neurons (neurovascular uncoupling), excessive formation of scar tissue (fibrosis), and the infiltration of detrimental substances from the bloodstream. Understanding these mechanisms of pericyte-driven demyelination may lead to the creation of new therapeutic strategies for tackling a range of neurodegenerative conditions.","PeriodicalId":501473,"journal":{"name":"ACS Pharmacology & Translational Science","volume":"7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142266167","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-17DOI: 10.1021/acsptsci.4c00293
Mohammad F. Khan, Mohammad M. Rahman, Yue Xin, Abdur Mustafa, Brian J. Smith, Karen M. Ottemann, Anna Roujeinikova
Quantification of protein–ligand interactions is crucial for understanding the protein’s biological function and for drug discovery. In this study, we employed three distinct approaches for determination of protein–ligand binding affinities by a thermal shift assay using a single ligand concentration. We present the results of the comparison of the performance of the conventional curve fitting (CF) method and two newly introduced methods - assuming zero heat capacity change across small temperature ranges (ZHC) and utilizing the unfolding equilibrium constant (UEC); the latter has the advantage of reducing calculations by obtaining the unfolding equilibrium constant directly from the experimental data. Our results highlight superior performance of the ZHC and UEC methods over the conventional CF method in estimating the binding affinity, irrespective of the ligand concentration. In addition, we evaluated how the new methods can be applied to high-throughput screening for potential binders, when the enthalpy (ΔHL) and molar heat capacity change (ΔCPL) of ligand binding are unknown. Our results suggest that, in this scenario, using the −300 cal K–1 mol–1 assumption for ΔCpL and either −5 kcal mol–1 or the average enthalpy efficiency-based estimation for ΔHL(T) can still provide reasonable estimates of the binding affinity. Incorporating the new methods into the workflow for screening of small drug-like molecules, typically conducted using single-concentration libraries, could greatly simplify and streamline the drug discovery process.
{"title":"Determination of Protein–Ligand Binding Affinities by Thermal Shift Assay","authors":"Mohammad F. Khan, Mohammad M. Rahman, Yue Xin, Abdur Mustafa, Brian J. Smith, Karen M. Ottemann, Anna Roujeinikova","doi":"10.1021/acsptsci.4c00293","DOIUrl":"https://doi.org/10.1021/acsptsci.4c00293","url":null,"abstract":"Quantification of protein–ligand interactions is crucial for understanding the protein’s biological function and for drug discovery. In this study, we employed three distinct approaches for determination of protein–ligand binding affinities by a thermal shift assay using a single ligand concentration. We present the results of the comparison of the performance of the conventional curve fitting (CF) method and two newly introduced methods - assuming zero heat capacity change across small temperature ranges (ZHC) and utilizing the unfolding equilibrium constant (UEC); the latter has the advantage of reducing calculations by obtaining the unfolding equilibrium constant directly from the experimental data. Our results highlight superior performance of the ZHC and UEC methods over the conventional CF method in estimating the binding affinity, irrespective of the ligand concentration. In addition, we evaluated how the new methods can be applied to high-throughput screening for potential binders, when the enthalpy (Δ<i>H</i><sub>L</sub>) and molar heat capacity change (Δ<i>C</i><sub>PL</sub>) of ligand binding are unknown. Our results suggest that, in this scenario, using the −300 cal K<sup>–1</sup> mol<sup>–1</sup> assumption for Δ<i>C</i><sub>pL</sub> and either −5 kcal mol<sup>–1</sup> or the average enthalpy efficiency-based estimation for Δ<i>H</i><sub>L</sub>(<i>T</i>) can still provide reasonable estimates of the binding affinity. Incorporating the new methods into the workflow for screening of small drug-like molecules, typically conducted using single-concentration libraries, could greatly simplify and streamline the drug discovery process.","PeriodicalId":501473,"journal":{"name":"ACS Pharmacology & Translational Science","volume":"25 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142266169","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-17DOI: 10.1021/acsptsci.4c00375
Elizabeth Grego, Sean M. Kelly, Jodi L. McGill, Michael Wannemuehler, Balaji Narasimhan
With limited therapies and vaccines available, human respiratory syncytial virus (HRSV) has a significant negative health impact on all age groups but particularly on infants, young children, and older adults. Bovine respiratory syncytial virus (BRSV) is pathogenically and antigenically similar to HRSV. Building upon previous studies using a BRSV nanovaccine coencapsulating multiple proteins, this work demonstrates the development and comparative evaluation of a coencapsulated nanovaccine to a cocktail nanovaccine formulation composed of polyanhydride nanoparticles encapsulating BRSV postfusion (F) glycoprotein and CpG ODN 1668 coadjuvant delivered simultaneously with nanoparticles encapsulating BRSV attachment glycoprotein (G) and CpG ODN 1668. These nanovaccine formulations were administered to C57BL/6 mice by one of two prime-boost regimens (i.e., intranasal/intranasal or intranasal/subcutaneous) followed by assessment of humoral immunity. The cocktail nanovaccine induced sustained anti-F and anti-G serum IgG antibody responses for 12 weeks postprimary immunization. Using polyanhydride particles to deliver G protein in a prime-boost regime also significantly induced serum anti-G antibodies compared to protein and coadjuvant alone. Serum IgG induced by the nanovaccine demonstrated virus-neutralizing capability from 42 to 119 days postprimary immunization. Further, anti-F IgG antibodies were detected in the bronchoalveolar lavage fluid of vaccinated animals. Finally, the nanovaccine induced long-lived anti-F antibody secreting plasma cells that were detectable in the bone marrow 205 days postprimary immunization. Overall, the BRSV nanovaccine(s) successfully induced long-lived humoral immune responses capable of virus neutralization, making this a promising vaccine candidate for further evaluation in other relevant animal models.
人类呼吸道合胞病毒（HRSV）的治疗方法和疫苗都很有限，它对所有年龄段的人群都有很大的负面健康影响，尤其是对婴儿、幼儿和老年人。牛呼吸道合胞病毒（BRSV）在致病性和抗原性上与 HRSV 相似。在之前使用包囊多种蛋白的 BRSV 纳米疫苗的研究基础上，本研究展示了包囊纳米疫苗与鸡尾酒纳米疫苗制剂的开发和比较评估，鸡尾酒纳米疫苗制剂由包囊 BRSV 融合后（F）糖蛋白和 CpG ODN 1668 佐剂的聚酸酐纳米颗粒与包囊 BRSV 附着糖蛋白（G）和 CpG ODN 1668 的纳米颗粒同时递送组成。C57BL/6 小鼠接种了这些纳米疫苗制剂，并采用了两种原位激活方案（即鼻内/鼻外或鼻内/皮下）中的一种，随后对体液免疫进行了评估。鸡尾酒纳米疫苗可在初次免疫后的12周内诱导持续的抗F和抗G血清IgG抗体反应。与单独使用蛋白质和辅助剂相比，使用多酸酐颗粒在原代增强机制中递送 G 蛋白也能显著诱导血清抗 G 抗体。纳米疫苗诱导的血清 IgG 在初次免疫后的 42 天至 119 天内都具有中和病毒的能力。此外，在接种动物的支气管肺泡灌洗液中也检测到了抗 F IgG 抗体。最后，纳米疫苗诱导了长效的抗 F 抗体分泌浆细胞，在初次免疫后 205 天的骨髓中可检测到这种浆细胞。总之，BRSV 纳米疫苗成功诱导了能够中和病毒的长效体液免疫反应，使其成为一种有希望在其他相关动物模型中进一步评估的候选疫苗。
{"title":"Bovine Respiratory Syncytial Virus Nanovaccine Induces Long-Lasting Humoral Immunity in Mice","authors":"Elizabeth Grego, Sean M. Kelly, Jodi L. McGill, Michael Wannemuehler, Balaji Narasimhan","doi":"10.1021/acsptsci.4c00375","DOIUrl":"https://doi.org/10.1021/acsptsci.4c00375","url":null,"abstract":"With limited therapies and vaccines available, human respiratory syncytial virus (HRSV) has a significant negative health impact on all age groups but particularly on infants, young children, and older adults. Bovine respiratory syncytial virus (BRSV) is pathogenically and antigenically similar to HRSV. Building upon previous studies using a BRSV nanovaccine coencapsulating multiple proteins, this work demonstrates the development and comparative evaluation of a coencapsulated nanovaccine to a cocktail nanovaccine formulation composed of polyanhydride nanoparticles encapsulating BRSV postfusion (F) glycoprotein and CpG ODN 1668 coadjuvant delivered simultaneously with nanoparticles encapsulating BRSV attachment glycoprotein (G) and CpG ODN 1668. These nanovaccine formulations were administered to C57BL/6 mice by one of two prime-boost regimens (i.e., intranasal/intranasal or intranasal/subcutaneous) followed by assessment of humoral immunity. The cocktail nanovaccine induced sustained anti-F and anti-G serum IgG antibody responses for 12 weeks postprimary immunization. Using polyanhydride particles to deliver G protein in a prime-boost regime also significantly induced serum anti-G antibodies compared to protein and coadjuvant alone. Serum IgG induced by the nanovaccine demonstrated virus-neutralizing capability from 42 to 119 days postprimary immunization. Further, anti-F IgG antibodies were detected in the bronchoalveolar lavage fluid of vaccinated animals. Finally, the nanovaccine induced long-lived anti-F antibody secreting plasma cells that were detectable in the bone marrow 205 days postprimary immunization. Overall, the BRSV nanovaccine(s) successfully induced long-lived humoral immune responses capable of virus neutralization, making this a promising vaccine candidate for further evaluation in other relevant animal models.","PeriodicalId":501473,"journal":{"name":"ACS Pharmacology & Translational Science","volume":"105 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142266168","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-13DOI: 10.1021/acsptsci.4c00348
Sylvain Auvity, Bala Attili, Fabien Caillé, Maud Goislard, Jérôme Cayla, Françoise Hinnen, Stéphane Demphel, Vincent Brulon, Michel Bottlaender, Claire Leroy, Guy Bormans, Bertrand Kuhnast, Marie-Anne Peyronneau
We have previously developed seven fluorinated analogues of A-836339 as new PET tracers for cannabinoid type 2 receptor (CB2R) imaging, among which (Z)-N-(3-(2-(2-[18F]fluoroethoxy)ethyl)-4,5-dimethylthiazol-2(3H)-ylidene)-2,2,3,3-tetramethylcyclopropane-1-carboxamide ([18F]FC0324) displayed high affinity and selectivity for CB2R in healthy rats. In the present study, we have further evaluated the imaging and metabolic properties of [18F]FC0324 in a rat model of human CB2R overexpression in the brain (AAV-hCB2) and in non-human primates (NHPs). Autoradiography with AAV-hCB2 rat brain sections exhibited a signal of [18F]FC0324 8-fold higher in the ipsilateral region than in the contralateral region. Blocking with NE40, a CB2R-specific agonist, resulted in a 91% decrease in the radioactivity. PET experiments showed a signal 7-fold higher in the ipsilateral region, and the specificity of [18F]FC0324 for hCB2R in vivo was confirmed by the 80% decrease after blocking with NE40. In NHPs, brain time-activity curves displayed a fast and homogeneous distribution followed by a rapid washout, in accordance with the low amount of CB2Rs in healthy brain. Whole-body PET-CT suggested a high and specific uptake of the radiotracer in the spleen, a CB2R-rich organ, and in the organs involved in metabolism and excretion, with a low bone uptake. In vitro metabolism with monkey liver microsomes (MLMs) led to the formation of six main hydroxylated metabolites of FC0324. Five of them were produced by human liver microsomes, being much less active than MLMs. In vivo, in NHPs, the main radiometabolite was likely to result from further oxidation of hydroxylated compounds, and parent [18F]FC0324 accounted for 8 ± 3% of plasma radioactivity (at 120 min p.i.) with a low level of potential interfering radiometabolites. Furthermore, this metabolism should be significantly reduced in humans due to species differences. In conclusion, [18F]FC0324 appears to be a promising candidate for further human studies with suitable kinetics, selectivity, and metabolic profile for CB2R PET imaging.
{"title":"Translational Preclinical PET Imaging and Metabolic Evaluation of a New Cannabinoid 2 Receptor (CB2R) Radioligand, (Z)-N-(3-(2-(2-[18F]Fluoroethoxy)ethyl)-4,5-dimethylthiazol-2(3H)-ylidene)-2,2,3,3-tetramethylcyclopropane-1-carboxamide","authors":"Sylvain Auvity, Bala Attili, Fabien Caillé, Maud Goislard, Jérôme Cayla, Françoise Hinnen, Stéphane Demphel, Vincent Brulon, Michel Bottlaender, Claire Leroy, Guy Bormans, Bertrand Kuhnast, Marie-Anne Peyronneau","doi":"10.1021/acsptsci.4c00348","DOIUrl":"https://doi.org/10.1021/acsptsci.4c00348","url":null,"abstract":"We have previously developed seven fluorinated analogues of A-836339 as new PET tracers for cannabinoid type 2 receptor (CB<sub>2</sub>R) imaging, among which (<i>Z</i>)-<i>N</i>-(3-(2-(2-[<sup>18</sup>F]fluoroethoxy)ethyl)-4,5-dimethylthiazol-2(3<i>H</i>)-ylidene)-2,2,3,3-tetramethylcyclopropane-1-carboxamide ([<sup>18</sup>F]FC0324) displayed high affinity and selectivity for CB<sub>2</sub>R in healthy rats. In the present study, we have further evaluated the imaging and metabolic properties of [<sup>18</sup>F]FC0324 in a rat model of human CB<sub>2</sub>R overexpression in the brain (AAV-<i>h</i>CB<sub>2</sub>) and in non-human primates (NHPs). Autoradiography with AAV-<i>h</i>CB<sub>2</sub> rat brain sections exhibited a signal of [<sup>18</sup>F]FC0324 8-fold higher in the ipsilateral region than in the contralateral region. Blocking with NE40, a CB<sub>2</sub>R-specific agonist, resulted in a 91% decrease in the radioactivity. PET experiments showed a signal 7-fold higher in the ipsilateral region, and the specificity of [<sup>18</sup>F]FC0324 for <i>h</i>CB<sub>2</sub>R in vivo was confirmed by the 80% decrease after blocking with NE40. In NHPs, brain time-activity curves displayed a fast and homogeneous distribution followed by a rapid washout, in accordance with the low amount of CB<sub>2</sub>Rs in healthy brain. Whole-body PET-CT suggested a high and specific uptake of the radiotracer in the spleen, a CB<sub>2</sub>R-rich organ, and in the organs involved in metabolism and excretion, with a low bone uptake. In vitro metabolism with monkey liver microsomes (MLMs) led to the formation of six main hydroxylated metabolites of FC0324. Five of them were produced by human liver microsomes, being much less active than MLMs. In vivo, in NHPs, the main radiometabolite was likely to result from further oxidation of hydroxylated compounds, and parent [<sup>18</sup>F]FC0324 accounted for 8 ± 3% of plasma radioactivity (at 120 min p.i.) with a low level of potential interfering radiometabolites. Furthermore, this metabolism should be significantly reduced in humans due to species differences. In conclusion, [<sup>18</sup>F]FC0324 appears to be a promising candidate for further human studies with suitable kinetics, selectivity, and metabolic profile for CB<sub>2</sub>R PET imaging.","PeriodicalId":501473,"journal":{"name":"ACS Pharmacology & Translational Science","volume":"9 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142266172","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-13DOI: 10.1021/acsptsci.4c00197
Anna Bérot, Ofelia Maniti, Saïd El Alaoui, Thierry Granjon, Meddy El Alaoui
Nanocarriers have found their interests in many fields including drug delivery and labeling of cells with the aim to target and eradicate tumor cells. One of the approaches to specifically address nanocarriers, such as liposomes, to their target is to attach antibodies of interest to their surface. To date, the development of immunoliposomes has been widely explored but has mainly involved chemical and unspecific reactions that could impair antibody stability, integrity, and orientation, thus reducing optimized immunoliposomes generation. In this study, we report the use of the patented COVISOLINK technology and the strain-promoted alkyne–azide cycloaddition (SPAAC) to generate immunoliposomes that target HER2 positive breast cancer with Trastuzumab as the antibody to be coupled. The efficacy of our two-step functionalization strategy and the successful specific coupling of the antibodies were validated by high-performance liquid chromatography-size exclusion chromatography (HPLC-SEC), which allowed a precise quantification of antibodies conjugated to liposomes and confirmed by cryo-TEM and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analyses. We also demonstrate by flow cytometry and epifluorescence microscopy that the produced anti-HER2 immunoliposomes were able to interact specifically with their target cells (SK-BR-3) while remaining negative with cells that express HER2 at a low level (MDA-MB-231). Hence, for the first time, our COVISOLINK strategy using microbial transglutaminase (mTG) enables the preparation and production of well-characterized immunoliposomes that could be used in different applications, including therapies.
{"title":"Generation of Anti-Epidermal Growth Factor Receptor-2 (HER2) Immunoliposomes Using Microbial Transglutaminase (mTG)-Mediated Site-Specific Conjugated Antibodies","authors":"Anna Bérot, Ofelia Maniti, Saïd El Alaoui, Thierry Granjon, Meddy El Alaoui","doi":"10.1021/acsptsci.4c00197","DOIUrl":"https://doi.org/10.1021/acsptsci.4c00197","url":null,"abstract":"Nanocarriers have found their interests in many fields including drug delivery and labeling of cells with the aim to target and eradicate tumor cells. One of the approaches to specifically address nanocarriers, such as liposomes, to their target is to attach antibodies of interest to their surface. To date, the development of immunoliposomes has been widely explored but has mainly involved chemical and unspecific reactions that could impair antibody stability, integrity, and orientation, thus reducing optimized immunoliposomes generation. In this study, we report the use of the patented COVISOLINK technology and the strain-promoted alkyne–azide cycloaddition (SPAAC) to generate immunoliposomes that target HER2 positive breast cancer with Trastuzumab as the antibody to be coupled. The efficacy of our two-step functionalization strategy and the successful specific coupling of the antibodies were validated by high-performance liquid chromatography-size exclusion chromatography (HPLC-SEC), which allowed a precise quantification of antibodies conjugated to liposomes and confirmed by cryo-TEM and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analyses. We also demonstrate by flow cytometry and epifluorescence microscopy that the produced anti-HER2 immunoliposomes were able to interact specifically with their target cells (SK-BR-3) while remaining negative with cells that express HER2 at a low level (MDA-MB-231). Hence, for the first time, our COVISOLINK strategy using microbial transglutaminase (mTG) enables the preparation and production of well-characterized immunoliposomes that could be used in different applications, including therapies.","PeriodicalId":501473,"journal":{"name":"ACS Pharmacology & Translational Science","volume":"5 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142266265","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Atherosclerosis is a major contributor to cardiovascular diseases with a high global prevalence. It is characterized by the formation of lipid-laden plaques in the arteries, which eventually lead to plaque rupture and thrombosis. While the current lipid-lowering therapies are generally effective in lowering the risk of cardiovascular events, they do not address the underlying causes of disease. Defective resolution of inflammation and impaired efferocytosis are the main driving forces of atherosclerosis. Macrophages recognize cells for clearance by the expression of “eat me” and “do not eat me” signals, including the CD47-SIRPα axis. However, the “do not eat me” signal CD47 is overexpressed in atherosclerotic plaques, leading to compromised efferocytosis and secondary necrosis. In this context, prophagocytic antibodies have been explored to stimulate the clearance of apoptotic cells, but they are nonspecific and impact healthy tissues. In macrophages, downstream of signal regulatory protein α, lie protein tyrosine phosphatases, SHP 1/2, which can serve as effective targets for selectively phagocytosing apoptotic cells. While increasing the efferocytosis targets the end stages of lesion development, the underlying issue of inflammation still persists. Simultaneously increasing efferocytosis and reducing inflammation can be effective therapeutic strategies for managing atherosclerosis. For instance, IL-10 is a key anti-inflammatory mediator that enhances efferocytosis via phosphoSTAT3 (pSTAT3) activation. In this study, we developed a combination nanotherapy by encapsulating an SHP-1 inhibitor (NSC 87877) and IL-10 in a single nanoparticle platform [(S + IL)-NPs] to enhance efferocytosis and inflammation resolution. Our studies suggest that (S + IL)-NPs successfully encapsulated both agents, entered the macrophages, and delivered the agents into intracellular compartments. Additionally, (S + IL)-NPs decreased inflammation by suppressing pro-inflammatory markers and enhancing anti-inflammatory mediators. They also exhibited the potential for improved phagocytic activity via pSTAT3 activation. Our nanomedicine-mediated upregulation of the anti-inflammatory and efferocytic responses in macrophages shows promise for the treatment of atherosclerosis.
{"title":"Development of Pro-resolving and Pro-efferocytic Nanoparticles for Atherosclerosis Therapy","authors":"Yukta Patel, Shireesha Manturthi, Saras Tiwari, Esha Gahunia, Amandine Courtemanche, Michelle Gandelman, Marceline Côté, Suresh Gadde","doi":"10.1021/acsptsci.4c00292","DOIUrl":"https://doi.org/10.1021/acsptsci.4c00292","url":null,"abstract":"Atherosclerosis is a major contributor to cardiovascular diseases with a high global prevalence. It is characterized by the formation of lipid-laden plaques in the arteries, which eventually lead to plaque rupture and thrombosis. While the current lipid-lowering therapies are generally effective in lowering the risk of cardiovascular events, they do not address the underlying causes of disease. Defective resolution of inflammation and impaired efferocytosis are the main driving forces of atherosclerosis. Macrophages recognize cells for clearance by the expression of “eat me” and “do not eat me” signals, including the CD47-SIRPα axis. However, the “do not eat me” signal CD47 is overexpressed in atherosclerotic plaques, leading to compromised efferocytosis and secondary necrosis. In this context, prophagocytic antibodies have been explored to stimulate the clearance of apoptotic cells, but they are nonspecific and impact healthy tissues. In macrophages, downstream of signal regulatory protein α, lie protein tyrosine phosphatases, SHP 1/2, which can serve as effective targets for selectively phagocytosing apoptotic cells. While increasing the efferocytosis targets the end stages of lesion development, the underlying issue of inflammation still persists. Simultaneously increasing efferocytosis and reducing inflammation can be effective therapeutic strategies for managing atherosclerosis. For instance, IL-10 is a key anti-inflammatory mediator that enhances efferocytosis via phosphoSTAT3 (pSTAT3) activation. In this study, we developed a combination nanotherapy by encapsulating an SHP-1 inhibitor (NSC 87877) and IL-10 in a single nanoparticle platform [(S + IL)-NPs] to enhance efferocytosis and inflammation resolution. Our studies suggest that (S + IL)-NPs successfully encapsulated both agents, entered the macrophages, and delivered the agents into intracellular compartments. Additionally, (S + IL)-NPs decreased inflammation by suppressing pro-inflammatory markers and enhancing anti-inflammatory mediators. They also exhibited the potential for improved phagocytic activity via pSTAT3 activation. Our nanomedicine-mediated upregulation of the anti-inflammatory and efferocytic responses in macrophages shows promise for the treatment of atherosclerosis.","PeriodicalId":501473,"journal":{"name":"ACS Pharmacology & Translational Science","volume":"33 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142266171","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-12DOI: 10.1021/acsptsci.4c00316
Linlin Li, Rongxi Wang, Li He, Hua Guo, Lei Fu, Guochang Wang, Jiarou Wang, Ziying Chen, Xingtong Peng, Xinyu Lu, Huimin Sui, Yuanyuan Jiang, Jie Zang, Lianghui Gao, Zhaohui Zhu
Angiotensin-converting enzyme 2 (ACE2) is not only a key to the renin–angiotensin–aldosterone system and related diseases, but also the main entry point on cell surfaces for certain coronaviruses, including severe acute respiratory syndrome coronavirus (SARS-CoV) and SARS-CoV-2. By analyzing the different key binding sites from the receptor-binding domain (RBD) of SARS-CoV and SARS-CoV-2, nine new ACE2-targeting peptides (A1 to A9) were designed, synthesized and connected with a chelator, 1,4,7-triazacyclononane-N,N′,N’’-triacetic acid (NOTA). NOTA-A1, NOTA-A2, NOTA-A4, NOTA-A5, and NOTA-A8 were successfully labeled with [68Ga]Ga3+ and were used for biological evaluation. [68Ga]Ga-NOTA-A2, [68Ga]Ga-NOTA-A5, and [68Ga]Ga-NOTA-A8 showed specific binding to ACE2 via cell assays, and their binding sites and binding capacity were calculated by molecular docking and molecular dynamics simulations. In tumor-bearing mice, A549 tumors were visualized 60 min postinjection of [68Ga]Ga-NOTA-A2, [68Ga]Ga-NOTA-A5, or [68Ga]Ga-NOTA-A8. These peptides also accumulated in the organs with high-level ACE2 expression, confirmed by immunohistochemical stain. Among them, [68Ga]Ga-NOTA-A5 exhibited the highest tumor uptake and tumor/background ratio, and it successfully tracked the increased ACE2 levels in mice tissues after excessive Losartan treatment. In a first-in-human study, the distribution of [68Ga]Ga-NOTA-A5 was evaluated with positron emission tomography/computed tomography (PET/CT) in three participants without adverse events. 68Ga-labeled peptides originated from the coronavirus RBD, with [68Ga]Ga-NOTA-A5 as a typical representative, seem to be safe and effective for the evaluation of ACE2 expression in vivo with PET/CT, facilitating further mechanism investigation and clinical evaluation of ACE2-related diseases.
{"title":"Evaluation of Angiotensin-Converting Enzyme 2 Expression In Vivo with Novel 68Ga-Labeled Peptides Originated from the Coronavirus Receptor-Binding Domain","authors":"Linlin Li, Rongxi Wang, Li He, Hua Guo, Lei Fu, Guochang Wang, Jiarou Wang, Ziying Chen, Xingtong Peng, Xinyu Lu, Huimin Sui, Yuanyuan Jiang, Jie Zang, Lianghui Gao, Zhaohui Zhu","doi":"10.1021/acsptsci.4c00316","DOIUrl":"https://doi.org/10.1021/acsptsci.4c00316","url":null,"abstract":"Angiotensin-converting enzyme 2 (ACE2) is not only a key to the renin–angiotensin–aldosterone system and related diseases, but also the main entry point on cell surfaces for certain coronaviruses, including severe acute respiratory syndrome coronavirus (SARS-CoV) and SARS-CoV-2. By analyzing the different key binding sites from the receptor-binding domain (RBD) of SARS-CoV and SARS-CoV-2, nine new ACE2-targeting peptides (A<b>1</b> to A<b>9</b>) were designed, synthesized and connected with a chelator, 1,4,7-triazacyclononane-<i>N,N′,N’</i>’-triacetic acid (NOTA). NOTA-A<b>1</b>, NOTA-A<b>2</b>, NOTA-A<b>4</b>, NOTA-A<b>5</b>, and NOTA-A<b>8</b> were successfully labeled with [<sup>68</sup>Ga]Ga<sup>3+</sup> and were used for biological evaluation. [<sup>68</sup>Ga]Ga-NOTA-A<b>2</b>, [<sup>68</sup>Ga]Ga-NOTA-A<b>5</b>, and [<sup>68</sup>Ga]Ga-NOTA-A<b>8</b> showed specific binding to ACE2 via cell assays, and their binding sites and binding capacity were calculated by molecular docking and molecular dynamics simulations. In tumor-bearing mice, A549 tumors were visualized 60 min postinjection of [<sup>68</sup>Ga]Ga-NOTA-A<b>2</b>, [<sup>68</sup>Ga]Ga-NOTA-A<b>5</b>, or [<sup>68</sup>Ga]Ga-NOTA-A<b>8</b>. These peptides also accumulated in the organs with high-level ACE2 expression, confirmed by immunohistochemical stain. Among them, [<sup>68</sup>Ga]Ga-NOTA-A<b>5</b> exhibited the highest tumor uptake and tumor/background ratio, and it successfully tracked the increased ACE2 levels in mice tissues after excessive Losartan treatment. In a first-in-human study, the distribution of [<sup>68</sup>Ga]Ga-NOTA-A<b>5</b> was evaluated with positron emission tomography/computed tomography (PET/CT) in three participants without adverse events. <sup>68</sup>Ga-labeled peptides originated from the coronavirus RBD, with [<sup>68</sup>Ga]Ga-NOTA-A<b>5</b> as a typical representative, seem to be safe and effective for the evaluation of ACE2 expression <i>in vivo</i> with PET/CT, facilitating further mechanism investigation and clinical evaluation of ACE2-related diseases.","PeriodicalId":501473,"journal":{"name":"ACS Pharmacology & Translational Science","volume":"11 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142218133","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-12DOI: 10.1021/acsptsci.4c00402
Jie Zhang, Sara S. Rinne, Wen Yin, Charles Dahlsson Leitao, Elvira Björklund, Ayman Abouzayed, Stefan Ståhl, John Löfblom, Anna Orlova, Torbjörn Gräslund, Anzhelika Vorobyeva
The outcome of clinical trials evaluating drugs targeting the human epidermal growth factor receptor 3 (HER3) has been poor, with primary concerns related to lack of efficacy. HER3 is considered a difficult target since its overexpression on tumors is relatively low and there is normal expression in many different organs. However, a significant number of patients across different cancer indications have overexpression of HER3 and the development of novel modalities targeting HER3 is therefore warranted. Here, we have investigated the properties of affibody-based drug conjugates targeting HER3. The HER3-targeting affibody molecule ZHER3 was fused in a mono- and bivalent format to an engineered albumin-binding domain (ABD) for in vivo half-life extension and was coupled to the cytotoxic drug DM1 via a non-cleavable maleimidocaproyl (mc) linker. In vivo, a moderate uptake was observed for [99mTc]Tc-labeled ZHER3-ABD-ZHER3-mcDM1 in HER3 expressing BxPC3 tumors (3.5 ± 0.3%IA/g) at 24 h after injection, and clearance was predominately renal-mediated. Treatment of mice with BxPC3 human pancreatic cancer xenografts showed that a combination of ZHER3-ABD-ZHER3-mcDM1 and its cytostatic analog ZHER3-ABD-ZHER3 was efficacious and superior to treatment with only ZHER3-ABD-ZHER3, providing tumor growth inhibition and longer median survival (90 d) in comparison to monotherapy (68 d) and vehicle control (49 d). ZHER3-ABD-ZHER3-mcDM1 was found to be a potent drug conjugate for the treatment of HER3-expressing tumors in mice.
{"title":"Affibody-Drug Conjugates Targeting the Human Epidermal Growth Factor Receptor-3 Demonstrate Therapeutic Efficacy in Mice Bearing Low Expressing Xenografts","authors":"Jie Zhang, Sara S. Rinne, Wen Yin, Charles Dahlsson Leitao, Elvira Björklund, Ayman Abouzayed, Stefan Ståhl, John Löfblom, Anna Orlova, Torbjörn Gräslund, Anzhelika Vorobyeva","doi":"10.1021/acsptsci.4c00402","DOIUrl":"https://doi.org/10.1021/acsptsci.4c00402","url":null,"abstract":"The outcome of clinical trials evaluating drugs targeting the human epidermal growth factor receptor 3 (HER3) has been poor, with primary concerns related to lack of efficacy. HER3 is considered a difficult target since its overexpression on tumors is relatively low and there is normal expression in many different organs. However, a significant number of patients across different cancer indications have overexpression of HER3 and the development of novel modalities targeting HER3 is therefore warranted. Here, we have investigated the properties of affibody-based drug conjugates targeting HER3. The HER3-targeting affibody molecule Z<sub>HER3</sub> was fused in a mono- and bivalent format to an engineered albumin-binding domain (ABD) for <i>in vivo</i> half-life extension and was coupled to the cytotoxic drug DM1 <i>via</i> a non-cleavable maleimidocaproyl (mc) linker. <i>In vivo</i>, a moderate uptake was observed for [<sup>99m</sup>Tc]Tc-labeled Z<sub>HER3</sub>-ABD-Z<sub>HER3</sub>-mcDM1 in HER3 expressing BxPC3 tumors (3.5 ± 0.3%IA/g) at 24 h after injection, and clearance was predominately renal-mediated. Treatment of mice with BxPC3 human pancreatic cancer xenografts showed that a combination of Z<sub>HER3</sub>-ABD-Z<sub>HER3</sub>-mcDM1 and its cytostatic analog Z<sub>HER3</sub>-ABD-Z<sub>HER3</sub> was efficacious and superior to treatment with only Z<sub>HER3</sub>-ABD-Z<sub>HER3</sub>, providing tumor growth inhibition and longer median survival (90 d) in comparison to monotherapy (68 d) and vehicle control (49 d). Z<sub>HER3</sub>-ABD-Z<sub>HER3</sub>-mcDM1 was found to be a potent drug conjugate for the treatment of HER3-expressing tumors in mice.","PeriodicalId":501473,"journal":{"name":"ACS Pharmacology & Translational Science","volume":"37 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142218144","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-12DOI: 10.1021/acsptsci.4c00265
Eleonora Panfili, Sarah Jane Rezzi, Annalisa Adamo, Daniele Mazzoletti, Alberto Massarotti, Riccardo Miggiano, Silvia Fallarini, Sara Ambrosino, Alice Coletti, Pasquale Molinaro, Michele Milella, Salvatore Paiella, Antonio Macchiarulo, Stefano Ugel, Tracey Pirali, Maria Teresa Pallotta
Indoleamine 2,3-dioxygenase 1 (IDO1) plays a key role in tumor immune escape. Besides being a metabolic enzyme that catalyzes the first step of tryptophan catabolism, it also acts as a signal-transducing protein, whose partnering with tyrosine phosphatase Src homology 2 (SH2) domain-containing protein tyrosine phosphatase substrate (SHPs) and phosphatidylinositol-3-kinase (PI3K) regulatory subunit p85 promotes the establishment of a sustained immunosuppressive phenotype. While IDO1 inhibitors typically interfere with its enzymatic activity, we aimed to discover a more effective modulator capable of blocking not only the enzymatic but also the signaling-mediated functions of IDO1. By virtual screening, we identified the compound VS-15, which selectively binds the heme-free form of IDO1, inhibits its enzymatic activity, and reduces the IDO1-mediated signaling pathway by negatively interfering with its partnership with SHPs and PI3K regulatory subunit p85 as well as with the IDO1 anchoring to the early endosomes in tumor cells. Moreover, VS-15 counteracts the TGF-β─mediated immunosuppressive phenotype in dendritic cells and reduces the level of inhibition of T cell proliferation by suppressive monocytes isolated from patients affected by pancreatic cancer. Herein, we describe the discovery and characterization of a small molecule with an unprecedented mechanism of action, capable of inhibiting both the enzymatic and nonenzymatic activities of IDO1 by binding to its apo-form. These results pave the way for the development of next-generation IDO1 inhibitors with a unique competitive advantage over the currently available modulators, thereby opening therapeutic opportunities in cancer immunotherapy.
{"title":"Identification of a Compound Inhibiting Both the Enzymatic and Nonenzymatic Functions of Indoleamine 2,3-Dioxygenase 1","authors":"Eleonora Panfili, Sarah Jane Rezzi, Annalisa Adamo, Daniele Mazzoletti, Alberto Massarotti, Riccardo Miggiano, Silvia Fallarini, Sara Ambrosino, Alice Coletti, Pasquale Molinaro, Michele Milella, Salvatore Paiella, Antonio Macchiarulo, Stefano Ugel, Tracey Pirali, Maria Teresa Pallotta","doi":"10.1021/acsptsci.4c00265","DOIUrl":"https://doi.org/10.1021/acsptsci.4c00265","url":null,"abstract":"Indoleamine 2,3-dioxygenase 1 (IDO1) plays a key role in tumor immune escape. Besides being a metabolic enzyme that catalyzes the first step of tryptophan catabolism, it also acts as a signal-transducing protein, whose partnering with tyrosine phosphatase Src homology 2 (SH2) domain-containing protein tyrosine phosphatase substrate (SHPs) and phosphatidylinositol-3-kinase (PI3K) regulatory subunit p85 promotes the establishment of a sustained immunosuppressive phenotype. While IDO1 inhibitors typically interfere with its enzymatic activity, we aimed to discover a more effective modulator capable of blocking not only the enzymatic but also the signaling-mediated functions of IDO1. By virtual screening, we identified the compound VS-15, which selectively binds the heme-free form of IDO1, inhibits its enzymatic activity, and reduces the IDO1-mediated signaling pathway by negatively interfering with its partnership with SHPs and PI3K regulatory subunit p85 as well as with the IDO1 anchoring to the early endosomes in tumor cells. Moreover, VS-15 counteracts the TGF-β─mediated immunosuppressive phenotype in dendritic cells and reduces the level of inhibition of T cell proliferation by suppressive monocytes isolated from patients affected by pancreatic cancer. Herein, we describe the discovery and characterization of a small molecule with an unprecedented mechanism of action, capable of inhibiting both the enzymatic and nonenzymatic activities of IDO1 by binding to its apo-form. These results pave the way for the development of next-generation IDO1 inhibitors with a unique competitive advantage over the currently available modulators, thereby opening therapeutic opportunities in cancer immunotherapy.","PeriodicalId":501473,"journal":{"name":"ACS Pharmacology & Translational Science","volume":"197 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142266170","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-09-11DOI: 10.1021/acsptsci.4c00372
Junwei Zhao, Monica Baiula, Elisabetta Cuna, Marco Francescato, Joanna Matalińska, Piotr F.J. Lipiński, Andrea Bedini, Luca Gentilucci
Recently, the fungus secondary metabolite cyclotetrapetide c[Trp-Phe-D-Pro-Phe] (CJ-15,208) and its derivatives deserved some attention for their unusual structure and distinctive in vitro and in vivo activity. These tryptophan-containing noncationic opioid peptides can be truly regarded as versatile picklocks capable of activating all opioid receptors. Intriguingly, minimal modification of the potent κ-opioid receptor (KOR) agonist c[D-Trp-Phe-Gly-β-Ala] (3) yielded c[D-Trp-Phe-β-Ala-β-Ala] (11), the first KOR-specific negative allosteric modulator (NAM) reported to-date. KOR exerts control over numerous functions in the central nervous system, including pain, depression, stress, mood, and reward. Hence, this KOR-selective NAM looks promising for modulating the KOR in addiction and neuropsychiatric disorders.
最近，真菌次生代谢物环四肽 c[Trp-Phe-D-Pro-Phe] （CJ-15,208）及其衍生物因其不同寻常的结构和独特的体内外活性而备受关注。这些含色氨酸的非阳离子类阿片肽可被真正视为能激活所有阿片受体的多功能螯合剂。有趣的是，对强效κ-阿片受体（KOR）激动剂c[D-Trp-Phe-Gly-β-Ala] (3)进行最小化修饰后，产生了c[D-Trp-Phe-β-Ala-β-Ala] (11)，这是迄今为止报道的首个KOR特异性负异调节剂（NAM）。KOR 可控制中枢神经系统的多种功能，包括疼痛、抑郁、压力、情绪和奖赏。因此，这种 KOR 选择性 NAM 对于调节成瘾和神经精神疾病中的 KOR 很有希望。
{"title":"Identification of c[D-Trp-Phe-β-Ala-β-Ala], the First κ-Opioid Receptor-Specific Negative Allosteric Modulator","authors":"Junwei Zhao, Monica Baiula, Elisabetta Cuna, Marco Francescato, Joanna Matalińska, Piotr F.J. Lipiński, Andrea Bedini, Luca Gentilucci","doi":"10.1021/acsptsci.4c00372","DOIUrl":"https://doi.org/10.1021/acsptsci.4c00372","url":null,"abstract":"Recently, the fungus secondary metabolite cyclotetrapetide c[Trp-Phe-D-Pro-Phe] (CJ-15,208) and its derivatives deserved some attention for their unusual structure and distinctive in vitro and in vivo activity. These tryptophan-containing noncationic opioid peptides can be truly regarded as versatile picklocks capable of activating all opioid receptors. Intriguingly, minimal modification of the potent κ-opioid receptor (KOR) agonist c[D-Trp-Phe-Gly-β-Ala] (<b>3</b>) yielded c[D-Trp-Phe-β-Ala-β-Ala] (<b>11</b>), the first KOR-specific negative allosteric modulator (NAM) reported to-date. KOR exerts control over numerous functions in the central nervous system, including pain, depression, stress, mood, and reward. Hence, this KOR-selective NAM looks promising for modulating the KOR in addiction and neuropsychiatric disorders.","PeriodicalId":501473,"journal":{"name":"ACS Pharmacology & Translational Science","volume":"10 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2024-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142218135","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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