Coronavirus disease 2019(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),is ongoing in human populations.According to the data fromtheWorld Health Organization(WHO),COVID-19 has resulte...Coronavirus disease 2019(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),is ongoing in human populations.According to the data fromtheWorld Health Organization(WHO),COVID-19 has resulted in 770 million confirmed cases and over 7 million deaths globally as of February 2025,profoundly impacting public health and socioeconomic development worldwide.展开更多
Background:The chicken chorioallantoic membrane(CAM)model is a potential alter-native to the mouse model based on the 3R principles.However,its value for deter-mination of the in vivo behaviors of radiolabeled peptide...Background:The chicken chorioallantoic membrane(CAM)model is a potential alter-native to the mouse model based on the 3R principles.However,its value for deter-mination of the in vivo behaviors of radiolabeled peptides through positron emission tomography(PET)imaging needed investigation.Herein,the chicken CAM tumor models were established,and their feasibility was evaluated for evaluating the imag-ing properties of radiolabeled peptides using a 68Ga-labeled HER2 affibody.Methods:Two human breast cancer cell lines were inoculated into chicken CAM and mice,respectively.The tumor-targeting potential and pharmacokinetic profile of a 68Ga-labeled affibody,68Ga-MZHER,in both tumor models were also determined.Results:The tumor-formation time in chicken CAM model was shorter than that of mouse model.The uptake values of human epithelial growth factor receptor-2(HER2)-positive Bcap37 tumors in chicken CAM and mouse models were 5.36±0.26%ID/g and 5.26±0.43%ID/g at 30 min postinjection of 68Ga-MZHER,respectively.At the same time points,the uptake values of HER2-negative MDA-MB-231 tumors in the chicken CAM models and mouse models were 1.57±0.15%ID/g and 1.67±0.25%ID/g,respectively.Ex vivo biodistribution confirmed that more radioactivity accu-mulated in Bcap37 tumors than in MDA-MD-231 tumors in both CAM and mouse models.Conclusion:In this study,the CAM tumor model was successfully prepared.The chicken CAM model is a novel tool for quickly determining the in vivo properties of radiolabeled peptides targeting biomarkers.It may be beneficial for early monitoring of the therapeutic effect of a new drug through PET imaging with specific peptides.展开更多
Affibody is a new class of small non-immunoglobulin affinity proteins that possesses high affinity at the picomole level to several tumor overexpressed receptors.Owing to the simple framework,affibody is flexible for ...Affibody is a new class of small non-immunoglobulin affinity proteins that possesses high affinity at the picomole level to several tumor overexpressed receptors.Owing to the simple framework,affibody is flexible for modification with payload,but the relatively low molecular weight of this construction simultaneously results in short half-life time which hinders its application in cancer therapy.In this work,we prepared a nanomedicine self-assembled from the conjugate of affibody(ZPDGFRβ:09591,PDGFRβ:platelet-derived growth factor receptorβ)with monomethyl auristatin E(MMAE)through cathepsin B cleavable dipeptide linker for targeted colorectal cancer therapy.The nanoscale characteristics of ZPDGFRβ:09591-MMAE affibody-drug conjugate nanomedicine(ZPDGFRβ:09591-M ADCN)resulted in enhanced pharmacokinetics,improved drug accumulation,and promoted biosecurity performance than those of free drugs.As a result,ZPDGFRβ:09591-M ADCN exhibited excellent antitumor efficacy with tumor inhibition rates(TIR)over 99.0%in PDGFRβ-positive tumor models with small solid tumors(~150 mm^(3))or large established tumors(~500 mm^(3)),indicating that ZPDGFRβ:09591-MMAE ADCN is promising for the clinic application in colorectal cancer therapy.展开更多
Follicular thyroid carcinoma(FTC)is the second most common form of thyroid malignancy,and it is associated with more aggressive growth and worse long-term survival outcomes relative to papillary thyroid carcinoma(PTC)...Follicular thyroid carcinoma(FTC)is the second most common form of thyroid malignancy,and it is associated with more aggressive growth and worse long-term survival outcomes relative to papillary thyroid carcinoma(PTC).Reliable approaches to preoperative FTC detection,however,remain to be established.Herein,a targeted Affibody-Au-Tripod nanoprobe was developed and successfully utilized to facilitate the targeted photoacoustic imaging(PAI)of epidermal growth factor receptor(EGFR)-positive cells and tumors.These Affibody-Au-Tripods were found to be highly sensitive and specific for cells expressing EGFR when used as a PA contrast agent in vitro,and studies conducted in an FTC-133 subcutaneous tumor model system in mice further revealed that these Affibody-Au-Tripods were able to specifically target these EGFR-expressing tumors while providing a strong photoacoustic signal in vivo.Importantly,these nanoprobes exhibited negligible cytotoxicity and robust chemical and physical stability,making Affibody-Au-Tripods promising candidates for targeted PAI-based FTC diagnosis.In addition,these nanoprobes have the potential to facilitate the individualized treatment of patients harboring EGFRpositive tumors.展开更多
Traditional development of small protein scaffolds has relied on display technologies and mutation-based engineering,which limit sequence and functional diversity,thereby constraining their therapeutic and application...Traditional development of small protein scaffolds has relied on display technologies and mutation-based engineering,which limit sequence and functional diversity,thereby constraining their therapeutic and application potential.Protein design tools have significantly advanced the creation of novel protein sequences,structures,and functions.However,further improvements in design strategies are still needed to more efficiently optimize the functional performance of protein-based drugs and enhance their druggability.Here,we extended an evolution-based design protocol to create a novel minibinder,BindHer,against the human epidermal growth factor receptor 2(HER2).It not only exhibits super stability and binding selectivity but also demonstrates remarkable properties in tissue specificity.Radiolabeling experiments with ^(99)mTc,^(68)Ga,and ^(18)F revealed that BindHer efficiently targets tumors in HER2-positive breast cancer mouse models,with minimal nonspecific liver absorption,outperforming scaffolds designed through traditional engineering.These findings highlight a new rational approach to automated protein design,offering significant potential for large-scale applications in therapeutic mini-protein development.展开更多
基金supported by the National Natural Science Foundation of China(grant number 32192452 to Y.S.)the National Key Research&Development Program of China(grant number 2023YFC2307800 to M.W.)+3 种基金the Beijing Life Science Academy(grant number 2023000CA0030 to Y.S.)the Major Project of Guangzhou National Laboratory(grant number GZNL2025C01041 to Y.S.)the National KeyR&D Program of China(grant number 2022YFC2303700 to Z.C.)Guangdong Basic and Applied Basic Research Foundation(grant number 2022B1515020059 to Z.C.).
文摘Coronavirus disease 2019(COVID-19),caused by severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),is ongoing in human populations.According to the data fromtheWorld Health Organization(WHO),COVID-19 has resulted in 770 million confirmed cases and over 7 million deaths globally as of February 2025,profoundly impacting public health and socioeconomic development worldwide.
基金This study was supported by the National Natural Science Foundation of China(grant numbers:31972644,32272959)the University Synergy Innovation Program of Anhui Province(grant number:GXXT-2019-035)+1 种基金the Jiangsu Provincial Medical Innovation Team(grant number:CXTDA2017024)the leading technology foundation research project of Jiangsu Province(grant number:BK20192005).
文摘Background:The chicken chorioallantoic membrane(CAM)model is a potential alter-native to the mouse model based on the 3R principles.However,its value for deter-mination of the in vivo behaviors of radiolabeled peptides through positron emission tomography(PET)imaging needed investigation.Herein,the chicken CAM tumor models were established,and their feasibility was evaluated for evaluating the imag-ing properties of radiolabeled peptides using a 68Ga-labeled HER2 affibody.Methods:Two human breast cancer cell lines were inoculated into chicken CAM and mice,respectively.The tumor-targeting potential and pharmacokinetic profile of a 68Ga-labeled affibody,68Ga-MZHER,in both tumor models were also determined.Results:The tumor-formation time in chicken CAM model was shorter than that of mouse model.The uptake values of human epithelial growth factor receptor-2(HER2)-positive Bcap37 tumors in chicken CAM and mouse models were 5.36±0.26%ID/g and 5.26±0.43%ID/g at 30 min postinjection of 68Ga-MZHER,respectively.At the same time points,the uptake values of HER2-negative MDA-MB-231 tumors in the chicken CAM models and mouse models were 1.57±0.15%ID/g and 1.67±0.25%ID/g,respectively.Ex vivo biodistribution confirmed that more radioactivity accu-mulated in Bcap37 tumors than in MDA-MD-231 tumors in both CAM and mouse models.Conclusion:In this study,the CAM tumor model was successfully prepared.The chicken CAM model is a novel tool for quickly determining the in vivo properties of radiolabeled peptides targeting biomarkers.It may be beneficial for early monitoring of the therapeutic effect of a new drug through PET imaging with specific peptides.
基金supported by the National Key Research and Development Plan of China(No.2020YFA0907702)the National Facility for Translational Medicine(Shanghai)(No.TMST-2020-001).
文摘Affibody is a new class of small non-immunoglobulin affinity proteins that possesses high affinity at the picomole level to several tumor overexpressed receptors.Owing to the simple framework,affibody is flexible for modification with payload,but the relatively low molecular weight of this construction simultaneously results in short half-life time which hinders its application in cancer therapy.In this work,we prepared a nanomedicine self-assembled from the conjugate of affibody(ZPDGFRβ:09591,PDGFRβ:platelet-derived growth factor receptorβ)with monomethyl auristatin E(MMAE)through cathepsin B cleavable dipeptide linker for targeted colorectal cancer therapy.The nanoscale characteristics of ZPDGFRβ:09591-MMAE affibody-drug conjugate nanomedicine(ZPDGFRβ:09591-M ADCN)resulted in enhanced pharmacokinetics,improved drug accumulation,and promoted biosecurity performance than those of free drugs.As a result,ZPDGFRβ:09591-M ADCN exhibited excellent antitumor efficacy with tumor inhibition rates(TIR)over 99.0%in PDGFRβ-positive tumor models with small solid tumors(~150 mm^(3))or large established tumors(~500 mm^(3)),indicating that ZPDGFRβ:09591-MMAE ADCN is promising for the clinic application in colorectal cancer therapy.
基金supported by the National Natural Science Foundation of China(81421004,81301268)Beijing Nova Program Interdisciplinary Cooperation Project (xxjc201812)+2 种基金International S&T Cooperation Program of China(2015DFA30440)Beijing Nova Program(Z131107000413063)CAMS Innovation Fund for Medical Sciences(CIFMS 2020-I2M-C&T-B-035)。
文摘Follicular thyroid carcinoma(FTC)is the second most common form of thyroid malignancy,and it is associated with more aggressive growth and worse long-term survival outcomes relative to papillary thyroid carcinoma(PTC).Reliable approaches to preoperative FTC detection,however,remain to be established.Herein,a targeted Affibody-Au-Tripod nanoprobe was developed and successfully utilized to facilitate the targeted photoacoustic imaging(PAI)of epidermal growth factor receptor(EGFR)-positive cells and tumors.These Affibody-Au-Tripods were found to be highly sensitive and specific for cells expressing EGFR when used as a PA contrast agent in vitro,and studies conducted in an FTC-133 subcutaneous tumor model system in mice further revealed that these Affibody-Au-Tripods were able to specifically target these EGFR-expressing tumors while providing a strong photoacoustic signal in vivo.Importantly,these nanoprobes exhibited negligible cytotoxicity and robust chemical and physical stability,making Affibody-Au-Tripods promising candidates for targeted PAI-based FTC diagnosis.In addition,these nanoprobes have the potential to facilitate the individualized treatment of patients harboring EGFRpositive tumors.
基金supported in part by the National Science and Technology Major Project(2019ZX09201003-003,China)the Key Research and Development Program of Sichuan Province(2020YFS0271,China)+2 种基金the National Natural Science Foundation of China(81973243)the Natural Science Foundation of Sichuan Province(2025ZNSFSC0663,China)the National University of Singapore(A-8001129-00-00).
文摘Traditional development of small protein scaffolds has relied on display technologies and mutation-based engineering,which limit sequence and functional diversity,thereby constraining their therapeutic and application potential.Protein design tools have significantly advanced the creation of novel protein sequences,structures,and functions.However,further improvements in design strategies are still needed to more efficiently optimize the functional performance of protein-based drugs and enhance their druggability.Here,we extended an evolution-based design protocol to create a novel minibinder,BindHer,against the human epidermal growth factor receptor 2(HER2).It not only exhibits super stability and binding selectivity but also demonstrates remarkable properties in tissue specificity.Radiolabeling experiments with ^(99)mTc,^(68)Ga,and ^(18)F revealed that BindHer efficiently targets tumors in HER2-positive breast cancer mouse models,with minimal nonspecific liver absorption,outperforming scaffolds designed through traditional engineering.These findings highlight a new rational approach to automated protein design,offering significant potential for large-scale applications in therapeutic mini-protein development.
