Neural EGFL-like 2(NELL2)is a secreted protein known for its regulatory functions in the nervous and reproductive systems,yet its role in bone biology remains unexplored.In this study,we observed that NELL2 was dimini...Neural EGFL-like 2(NELL2)is a secreted protein known for its regulatory functions in the nervous and reproductive systems,yet its role in bone biology remains unexplored.In this study,we observed that NELL2 was diminished in the bone of aged and ovariectomized(OVX)mice,as well as in the serum of osteopenia and osteoporosis patients.In vitro loss-of-function and gain-offunction studies revealed that NELL2 facilitated osteoblast differentiation and impeded adipocyte differentiation from stromal progenitor cells.In vivo studies further demonstrated that the deletion of NELL2 in preosteoblasts resulted in decreased cancellous bone mass in mice.Mechanistically,NELL2 interacted with the FNI-type domain located at the C-terminus of Fibronectin 1(Fn1).Moreover,we found that NELL2 activated the focal adhesion kinase(FAK)/AKT signaling pathway through Fn1/integrinβ1(ITGB1),leading to the promotion of osteogenesis and the inhibition of adipogenesis.Notably,administration of NELL2-AAV was found to ameliorate bone loss in OVX mice.These findings underscore the significant role of NELL2 in osteoblast differentiation and bone homeostasis,suggesting its potential as a therapeutic target for managing osteoporosis.展开更多
Fibronectin glomerulopathy is a rare autosomal dominant inherited glomerular disease associated with massive deposition of fibronectin.We recently diagnosed fibronectin glomerulopathy in a 29-year-old woman presenting...Fibronectin glomerulopathy is a rare autosomal dominant inherited glomerular disease associated with massive deposition of fibronectin.We recently diagnosed fibronectin glomerulopathy in a 29-year-old woman presenting nephrotic syndrome.Genetic analysis of fibronectin 1 gene showed heterozygosity for the Y973C mutation.However,this mutation was not found in her parents.She had stable renal function but persistent nephrotic proteinuria after one-year follow-up.展开更多
An understanding of osteoblast adhesion and proliferation on biomaterials is crucial to optimizing the surfaces of artificial implants used in clinical practice. Polished, anodic oxidation （AO） and micro-arc oxidati...An understanding of osteoblast adhesion and proliferation on biomaterials is crucial to optimizing the surfaces of artificial implants used in clinical practice. Polished, anodic oxidation （AO） and micro-arc oxidation （MAO） treated titanium （Ti） plates were used as model surfaces to study the adhesion of MG-63 cells. Cells were monitored for 0.5 and 4 h; faster adhesion and spreading of MG-63 ceils were observed on the AO and MAO modified samples. Stimulated secretion of fibronectin （FN） influenced the adhesion rates. In addition, AO and MAO modified surfaces promoted cell proliferation through apparent up-regulation of FN and integrin a5 transcription via outside-in signaling. This strongly suggests that FN secretion by osteoblasts plays an essential role in enhanced cell adhesion, spreading and proliferation on these modified Ti surfaces.展开更多
Objective:While cisplatin-based chemotherapy is pivotal for advanced bladder cancer,acquired resistance remains a major obstacle.This study investigates key molecular drivers of this resistance and potential reversal ...Objective:While cisplatin-based chemotherapy is pivotal for advanced bladder cancer,acquired resistance remains a major obstacle.This study investigates key molecular drivers of this resistance and potential reversal strategies.Methods:We established GC(Gemcitabine and Cisplatin)-resistant T24-R and UC3-R cell lines from T24 and UM-UC-3(UC3)cells.Transcriptomic and proteomic analyses identified differentially expressed molecules.Apoptosis and cell viability were assessed by flow cytometry and CCK-8(Cell Counting Kit-8)assays,while RT-qPCR(Reverse Transcription Quantitative Polymerase Chain Reaction)and Western blot analyzed gene and protein expression.Immunofluorescence evaluated FAK(Focal Adhesion Kinase)phosphorylation,and a xenograft mouse model validated the findings in vivo.Results:Integrated transcriptomic and proteomic analysis identified FN1(fibronectin)as a consistently upregulated top candidate in resistant cells(T24-R transcript log_(2)FC=2.8,protein log_(2)FC=0.9;UC3-R transcript log_(2)FC=3.7;all p<0.001).Knockdown of FN1 reduced chemoresistance(Resistance Index:5.2 in T24-R and 2.0 in UC3-R cells,p<0.001)and enhanced apoptosis(approximately 4.5-fold in T24-R and 7.5-fold in UC3-R,p<0.001).ITGB4(Integrin Subunit Beta 4)was upregulated in resistant cells(transcript log_(2)FC:4.2 in T24-R and 3.03 in UC3-R;protein log_(2)FC:0.67 in T24-R;all p<0.01).Critically,ITGB4 knockdown abolished the chemoresistance promoted by exogenous FN1,which was associated with increased FAK(Y397)phosphorylation.Conclusion:Our results demonstrate that the FN1-ITGB4 axis drives chemoresistance in bladder cancer via FAK signaling.Targeting this axis represents a promising strategy to overcome chemoresistance.展开更多
miRNAs play important roles in the post-transcriptional regulation of most transcripts and control several biological processes. We have shown that miR-200b regulates VEGF and miR-146a regulates ECM （extracellular ma...miRNAs play important roles in the post-transcriptional regulation of most transcripts and control several biological processes. We have shown that miR-200b regulates VEGF and miR-146a regulates ECM （extracellular matrix） protein, FN （fibronectin） production. We examined the effects of these two miRNAs in wound healing in vitro and in animals with or without diabetes. Microvascular ECs （endothelial cells） with or without miR inhibitor （antagomir） transfection were tested for VEGF and FN production. Scratch assays and angiogenesis assays were performed. Wounds in the back of mice with or without streptozotocin-induced diabetes were treated with antagomirs alone or in combinations. The wounds were measured and tissues were examined for mRNAs, proteins and miRNAs and examined histologically. In the ECs, miR-200b or miR-146a inhibitors increased VEGF and FN production, cell migration and tube formation. Wound healing in the animals, along with increased production of specific proteins were accelerated by miR-200b or miR-146a inhibitor treatment and was pronounced when these two were combined In diabetes, although delayed, similar patterns were seen. These results indicated that combination of miR-146a and miR-200b inhibitor treatment is useful in wound healing both in normal and in diabetic conditions, miRNA mediated wound healing may potentially constitute a novel therapeutic approach.展开更多
基金supported by grants from National Natural Science Foundation of China(82272444,81972031,81972033)China Postdoctoral Science Foundation(2022M722382)Tianjin Key Medical Discipline(Specialty)Construction Project(TJYXZDXK-032A)。
文摘Neural EGFL-like 2(NELL2)is a secreted protein known for its regulatory functions in the nervous and reproductive systems,yet its role in bone biology remains unexplored.In this study,we observed that NELL2 was diminished in the bone of aged and ovariectomized(OVX)mice,as well as in the serum of osteopenia and osteoporosis patients.In vitro loss-of-function and gain-offunction studies revealed that NELL2 facilitated osteoblast differentiation and impeded adipocyte differentiation from stromal progenitor cells.In vivo studies further demonstrated that the deletion of NELL2 in preosteoblasts resulted in decreased cancellous bone mass in mice.Mechanistically,NELL2 interacted with the FNI-type domain located at the C-terminus of Fibronectin 1(Fn1).Moreover,we found that NELL2 activated the focal adhesion kinase(FAK)/AKT signaling pathway through Fn1/integrinβ1(ITGB1),leading to the promotion of osteogenesis and the inhibition of adipogenesis.Notably,administration of NELL2-AAV was found to ameliorate bone loss in OVX mice.These findings underscore the significant role of NELL2 in osteoblast differentiation and bone homeostasis,suggesting its potential as a therapeutic target for managing osteoporosis.
文摘Fibronectin glomerulopathy is a rare autosomal dominant inherited glomerular disease associated with massive deposition of fibronectin.We recently diagnosed fibronectin glomerulopathy in a 29-year-old woman presenting nephrotic syndrome.Genetic analysis of fibronectin 1 gene showed heterozygosity for the Y973C mutation.However,this mutation was not found in her parents.She had stable renal function but persistent nephrotic proteinuria after one-year follow-up.
基金Project (2010DFA32270) supported by the Ministry of Science and Technology of China (International Science & Technology Cooperation Program of China) Project (51102090) supported by the National Natural Science Foundation of ChinaProject (NCET-12-0170) supported by the Program for New Century Excellent Talents in University of China
文摘An understanding of osteoblast adhesion and proliferation on biomaterials is crucial to optimizing the surfaces of artificial implants used in clinical practice. Polished, anodic oxidation （AO） and micro-arc oxidation （MAO） treated titanium （Ti） plates were used as model surfaces to study the adhesion of MG-63 cells. Cells were monitored for 0.5 and 4 h; faster adhesion and spreading of MG-63 ceils were observed on the AO and MAO modified samples. Stimulated secretion of fibronectin （FN） influenced the adhesion rates. In addition, AO and MAO modified surfaces promoted cell proliferation through apparent up-regulation of FN and integrin a5 transcription via outside-in signaling. This strongly suggests that FN secretion by osteoblasts plays an essential role in enhanced cell adhesion, spreading and proliferation on these modified Ti surfaces.
基金supported by grants from the National Natural Science Foundation of China(82372881 to Weiyang He)the Chongqing Biomedicine Key R&D Project(CSTB2021TIAD-KPX0041 to Weiyang He).
文摘Objective:While cisplatin-based chemotherapy is pivotal for advanced bladder cancer,acquired resistance remains a major obstacle.This study investigates key molecular drivers of this resistance and potential reversal strategies.Methods:We established GC(Gemcitabine and Cisplatin)-resistant T24-R and UC3-R cell lines from T24 and UM-UC-3(UC3)cells.Transcriptomic and proteomic analyses identified differentially expressed molecules.Apoptosis and cell viability were assessed by flow cytometry and CCK-8(Cell Counting Kit-8)assays,while RT-qPCR(Reverse Transcription Quantitative Polymerase Chain Reaction)and Western blot analyzed gene and protein expression.Immunofluorescence evaluated FAK(Focal Adhesion Kinase)phosphorylation,and a xenograft mouse model validated the findings in vivo.Results:Integrated transcriptomic and proteomic analysis identified FN1(fibronectin)as a consistently upregulated top candidate in resistant cells(T24-R transcript log_(2)FC=2.8,protein log_(2)FC=0.9;UC3-R transcript log_(2)FC=3.7;all p<0.001).Knockdown of FN1 reduced chemoresistance(Resistance Index:5.2 in T24-R and 2.0 in UC3-R cells,p<0.001)and enhanced apoptosis(approximately 4.5-fold in T24-R and 7.5-fold in UC3-R,p<0.001).ITGB4(Integrin Subunit Beta 4)was upregulated in resistant cells(transcript log_(2)FC:4.2 in T24-R and 3.03 in UC3-R;protein log_(2)FC:0.67 in T24-R;all p<0.01).Critically,ITGB4 knockdown abolished the chemoresistance promoted by exogenous FN1,which was associated with increased FAK(Y397)phosphorylation.Conclusion:Our results demonstrate that the FN1-ITGB4 axis drives chemoresistance in bladder cancer via FAK signaling.Targeting this axis represents a promising strategy to overcome chemoresistance.
文摘miRNAs play important roles in the post-transcriptional regulation of most transcripts and control several biological processes. We have shown that miR-200b regulates VEGF and miR-146a regulates ECM （extracellular matrix） protein, FN （fibronectin） production. We examined the effects of these two miRNAs in wound healing in vitro and in animals with or without diabetes. Microvascular ECs （endothelial cells） with or without miR inhibitor （antagomir） transfection were tested for VEGF and FN production. Scratch assays and angiogenesis assays were performed. Wounds in the back of mice with or without streptozotocin-induced diabetes were treated with antagomirs alone or in combinations. The wounds were measured and tissues were examined for mRNAs, proteins and miRNAs and examined histologically. In the ECs, miR-200b or miR-146a inhibitors increased VEGF and FN production, cell migration and tube formation. Wound healing in the animals, along with increased production of specific proteins were accelerated by miR-200b or miR-146a inhibitor treatment and was pronounced when these two were combined In diabetes, although delayed, similar patterns were seen. These results indicated that combination of miR-146a and miR-200b inhibitor treatment is useful in wound healing both in normal and in diabetic conditions, miRNA mediated wound healing may potentially constitute a novel therapeutic approach.
