Reproductive hormones associated with the hypothalamic-pituitary-gonadal(HPG)axis are closely linked to bone homeostasis.In this study,we demonstrate that Gonadotropin inhibitory hormone(GnIH,one of the key reproducti...Reproductive hormones associated with the hypothalamic-pituitary-gonadal(HPG)axis are closely linked to bone homeostasis.In this study,we demonstrate that Gonadotropin inhibitory hormone(GnIH,one of the key reproductive hormones upstream of the HPG axis)plays an indispensable role in regulating bone homeostasis and maintaining bone mass.We find that deficiency of GnIH or its receptor Gpr147 leads to a significant reduction in bone mineral density(BMD)in mice primarily by enhancement of osteoclast activation in vivo and in vitro.Mechanistically,GnIH/Gpr147 inhibits osteoclastogenesis by the PI3K/AKT,MAPK,NF-κB and Nfatc1 signaling pathways.Furthermore,GnIH treatment was able to alleviate bone loss in aging,ovariectomy(OVX)or LPS-induced mice.Moreover,the therapy using green light promotes the release of GnIH and rescues OVX-induced bone loss.In humans,serum GnIH increases and bone resorption markers decrease after green light exposure.Therefore,our study elucidates that GnIH plays an important role in maintaining bone homeostasis via modulating osteoclast differentiation and demonstrates the potential of GnIH therapy or green light therapy in preventing osteoporosis.展开更多
Prostaglandin E2(PGE2), a major cyclooxygenase-2(COX-2) product, is highly secreted by the osteoblast lineage in the subchondral bone tissue of osteoarthritis(OA) patients. However, NSAIDs, including COX-2 inhibitors,...Prostaglandin E2(PGE2), a major cyclooxygenase-2(COX-2) product, is highly secreted by the osteoblast lineage in the subchondral bone tissue of osteoarthritis(OA) patients. However, NSAIDs, including COX-2 inhibitors, have severe side effects during OA treatment. Therefore, the identification of novel drug targets of PGE2 signaling in OA progression is urgently needed. Osteoclasts play a critical role in subchondral bone homeostasis and OA-related pain. However, the mechanisms by which PGE2 regulates osteoclast function and subsequently subchondral bone homeostasis are largely unknown. Here, we show that PGE2 acts via EP4 receptors on osteoclasts during the progression of OA and OA-related pain. Our data show that while PGE2 mediates migration and osteoclastogenesis via its EP2 and EP4 receptors, tissue-specific knockout of only the EP4 receptor in osteoclasts(EP4 Lys M) reduced disease progression and osteophyte formation in a murine model of OA. Furthermore, OA-related pain was alleviated in the EP4 Lys M mice, with reduced Netrin-1 secretion and CGRP-positive sensory innervation of the subchondral bone. The expression of plateletderived growth factor-BB(PDGF-BB) was also lower in the EP4 Lys Mmice, which resulted in reduced type H blood vessel formation in subchondral bone. Importantly, we identified a novel potent EP4 antagonist, HL-43, which showed in vitro and in vivo effects consistent with those observed in the EP4 Lys Mmice. Finally, we showed that the Gαs/PI3 K/AKT/MAPK signaling pathway is downstream of EP4 activation via PGE2 in osteoclasts. Together, our data demonstrate that PGE2/EP4 signaling in osteoclasts mediates angiogenesis and sensory neuron innervation in subchondral bone, promoting OA progression and pain, and that inhibition of EP4 with HL-43 has therapeutic potential in OA.展开更多
The superfamily of G protein-coupled receptors (GPCRs) contains immense structural and functional diversity and mediates a myriad of biological processes upon activation by various extracellular signals.Critical roles...The superfamily of G protein-coupled receptors (GPCRs) contains immense structural and functional diversity and mediates a myriad of biological processes upon activation by various extracellular signals.Critical roles of GPCRs have been established in bone development,remodeling,and disease.Multiple human GPCR mutations impair bone development or metabolism,resulting in osteopathologies.Here we summarize the disease phenotypes and dysfunctions caused by GPCR gene mutations in humans as well as by deletion in animals.To date,92 receptors (5 glutamate family,67 rhodopsin family,5 adhesion,4 frizzled/taste2 family,5 secretin family,and 6 other 7TM receptors) have been associated with bone diseases and dysfunctions (36 in humans and 72 in animals).By analyzing data from these 92 GPCRs,we found that mutation or deletion of different individual GPCRs could induce similar bone diseases or dysfunctions,and the same individual GPCR mutation or deletion could induce different bone diseases or dysfunctions in different populations or animal models.Data from human diseases or dysfunctions identified 19 genes whose mutation was associated with human BMD:9 genes each for human height and osteoporosis;4 genes each for human osteoarthritis (OA) and fracture risk;and 2 genes each for adolescent idiopathic scoliosis (AIS),periodontitis,osteosarcoma growth,and tooth development.Reports from gene knockout animals found 40 GPCRs whose deficiency reduced bone mass,while deficiency of 22 GPCRs increased bone mass and BMD;deficiency of 8 GPCRs reduced body length,while 5 mice had reduced femur size upon GPCR deletion.Furthermore,deficiency in 6 GPCRs induced osteoporosis;4 induced osteoarthritis;3 delayed fracture healing;3 reduced arthritis severity;and reduced bone strength,increased bone strength,and increased cortical thickness were each observed in 2 GPCR-deficiency models.The ever-expanding number of GPCR mutation-associated diseases warrants accelerated molecular analysis,population studies,and investigation of phenotype correlation with SNPs to elucidate GPCR function in human diseases.展开更多
AIM: To investigate the role of claudin 1 in the regulation of genes involved in cell migration and tumor necrosis factor alpha (TNF-α)-induced gene expression in human gastric adenocarcinoma cells.
Prostate cancer (PCa) is the most common male cancer [1, 2]. PCa initially depends on androgen receptor (AR) signaling for growth and survival. Androgen deprivation therapy causes a temporary reduction in PCa tumor bu...Prostate cancer (PCa) is the most common male cancer [1, 2]. PCa initially depends on androgen receptor (AR) signaling for growth and survival. Androgen deprivation therapy causes a temporary reduction in PCa tumor burden, but the tumor eventually develops into castrationresistant prostate cancer (CRPC) with the ability to grow again in the absence of androgens [3]. Mechanisms of CRPC progression include AR amplification and overexpression [4], AR gene rearrangement promoting synthesis of constitutively-active truncated AR splice variants (ARVs) [4], and induction of intracrine androgen metabolic enzymes [3]. Current anti-androgen therapies including MDV3100 (Enzalutamide) and abiraterone have focused on the androgen-dependent activation of AR through its ligand-binding domain (LBD), but do not provide a continuing clinical benefit for patients with CRPC and presumably fail due to multiple mechanisms including the expression of AR-Vs lacking the LBD [5]. These AR-Vs signal in the absence of ligand and are therefore resistant to LBD-targeting AR antagonists or agents that repress androgen biosynthesis [6].展开更多
Use of gold nanoparticles(GNPs)in medicine is an emerging field of translational research with vast clinical implications and exciting therapeutic potential.However,the safety of using GNPs in human subjects is an imp...Use of gold nanoparticles(GNPs)in medicine is an emerging field of translational research with vast clinical implications and exciting therapeutic potential.However,the safety of using GNPs in human subjects is an important question that remains unanswered.This study reviews over 20 clinical trials focused on GNP safety and aims to summarize all the clinical studies,completed and ongoing,to identify whether GNPs are safe to use in humans as a therapeutic platform.In these studies,GNPs were implemented as drug delivery devices,for photothermal therapy,and utilized for their intrinsic therapeutic effects by various routes of delivery.These studies revealed no major safety concerns with the use of GNPs;however,the number of trials and total patient number remains limited.Multi-dose,multicenter blinded trials are required to deepen our understanding of the use of GNPs in clinical settings to facilitate translation of this novel,multifaceted therapeutic device.Expanding clinical trials will require collaboration between clinicians,scientists,and biotechnology companies.展开更多
CRISPR/Cas9-mediated site-specific insertion of exogenous genes holds potential for clinical applications.However,it is still infeasible because homologous recombination(HR)is inefficient,especially for nondividing ce...CRISPR/Cas9-mediated site-specific insertion of exogenous genes holds potential for clinical applications.However,it is still infeasible because homologous recombination(HR)is inefficient,especially for nondividing cells.To overcome the challenge,we report that a homology-independent targeted integration(HITI)strategy is used for permanent integration of high-specificity-activity Factor IX variant(F9 Padua,R338L)at the albumin(Alb)locus in a novel hemophilia B(HB)rat model.The knock-in efficiency reaches 3.66%,as determined by droplet digital PCR(dd PCR).The clotting time is reduced to a normal level four weeks after treatment,and the circulating factor IX(FIX)level is gradually increased up to 52%of the normal level over nine months even after partial hepatectomy,demonstrating the amelioration of hemophilia.Through primer-extension-mediated sequencing(PEM-seq),no significant off-target effect is detected.This study not only provides a novel model for HB but also identifies a promising therapeutic approach for rare inherited diseases.展开更多
The KRAS gene is frequently mutated in multiple cancer types,but it fell off the drug discovery radar for many years because of its inherent "undruggable" structure and undefined biological properties.As rep...The KRAS gene is frequently mutated in multiple cancer types,but it fell off the drug discovery radar for many years because of its inherent "undruggable" structure and undefined biological properties.As reported in the paper entitled "Suppression of KRas-mutant cancer through the combined inhibition of KRAS with PLK1 and ROCK" in Nature Communications,we performed a synthetic lethal screening with a combinatorial strategy on a panel of clinical drugs;we found that combined inhibition of polo-like kinase 1 and RhoA/Rho kinase markedly suppressed tumor growth in mice.An increase in the expression of the tumor suppressor P21^(WAF1/CIP1) contributed to the synergistic mechanism of the combination therapy.These findings open a novel avenue for the treatment of KRAS-mutant lung cancer.展开更多
Energy storage is pivotal for the continuous utilization of solar energy suffering from the intermittency issue. Herein, we demonstrate a solar rechargeable flow cell（SRFC） based on photoelectrochemical regeneration...Energy storage is pivotal for the continuous utilization of solar energy suffering from the intermittency issue. Herein, we demonstrate a solar rechargeable flow cell（SRFC） based on photoelectrochemical regeneration of vanadium redox species for in-situ solar energy harvest and storage. In this device, TiO_2 and MWCNT/acetylene black（MWCNT/AB） composite are served as the photoanode and the counter electrode,respectively, with all vanadium redox couples, VO_2~＋/VO^（2＋）and VO^（2＋）/V^（3＋）, as solar energy storage media.Benefitting from solar energy, the cell can be photocharged under a bias as low as 0.1 V, which is much lower than the discharge voltage of ~0.5 V. Photocharged under the optimized condition, the cell delivers a discharge energy of 23.0 mWh/L with 67.4% input electric energy savings. This prototype work may inspire the rational design for cost-effective solar energy storage devices.展开更多
The authors regret that the grant number“21CJ1402200”in the Acknowledgments session should be replaced as“21JC1402200”.The corrected contents areprovided as follows.
Glioblastoma multiforme(GBM)is the deadliest form of brain tumor,and effective treatments are lacking.Thus,a new generation of effective treatments is urgently needed.B-cell lymphoma 6(BCL6)is a transcription factor t...Glioblastoma multiforme(GBM)is the deadliest form of brain tumor,and effective treatments are lacking.Thus,a new generation of effective treatments is urgently needed.B-cell lymphoma 6(BCL6)is a transcription factor that functions to suppress the transcription of DNA damage response genes,halting cell death in response to DNA damage.Here,we identified BCL6 as a lynchpin in GBM,the expression of which was greater in GBM cells than in normal cells and associated with poor survival in GBM patients.The silencing of BCL6 additionally affected GBM cell proliferation and triggered cellular damage.Furthermore,we reported the identification of YK01,a novel small-molecule inhibitor of BCL6.YK01 exhibited excellent anti-GBM bioactivity and caused apoptosis;importantly,YK01 significantly inhibited the growth of GBM cells both in vitro and in vivo.Moreover,the combination of YK01 and temozolomide treatment significantly suppressed the growth and metastasis of tumors in vivo and prolonged the survival of mice with tumors.In summary,our findings reveal that BCL6 appears to play a crucial role in GBM and may be a therapeutic target for treating this incurable condition.展开更多
In target-based drug design,the manual creation of a poor initial compound library,the time-consuming wetlaboratory experimental screening method,and the weak explainability of their activity against compounds signifi...In target-based drug design,the manual creation of a poor initial compound library,the time-consuming wetlaboratory experimental screening method,and the weak explainability of their activity against compounds significantly limit the efficiency of discovering novel therapeutics.Here we propose an image-guided,interpretability deep learning workflow,named LeadDisFlow,to enable rapid,accurate target drug discovery.Using LeadDisFlow,we identified four potent antagonists with single-nanomolar antagonistic activity against PGE2 receptor subtype 4(EP4),a promising target for tumor im-munotherapy.Remarkably,the most potent EP4 antagonist,ZY001,demonstrated an IC50 value of(0.51±0.02)nM,along with high selectivity.Furthermore,ZY001 effectively impaired the PGE2-induced gene expression of a panel of immunosuppressive molecules in macrophages.The workflow facilitates the discovery of potent EP4 antagonists that enhance anti-tumor immune response,and provides a convenient and quick approach to discover promising therapeutics for a specific drug target.展开更多
CRISPR/Cas9 is a versatile genome-editing tool which is widely used for modifying the genome of both prokaryotic and eukaryotic organisms for basic research and applications. An increasing number of reports have demon...CRISPR/Cas9 is a versatile genome-editing tool which is widely used for modifying the genome of both prokaryotic and eukaryotic organisms for basic research and applications. An increasing number of reports have demonstrated that CRISPR/Cas9-mediated genome editing is a powerful technology for gene therapy. Here, we review the recent advances in CRISPR/Cas9-mediated gene therapy in animal models via different strategies and discuss the challenges as well as future prospects.展开更多
Osteoarthritis(OA)is one of the most common chronic diseases in the world.However,current treatment modalities mainly relieve pain and inhibit cartilage degradation,but do not promote cartilage regeneration.In this st...Osteoarthritis(OA)is one of the most common chronic diseases in the world.However,current treatment modalities mainly relieve pain and inhibit cartilage degradation,but do not promote cartilage regeneration.In this study,we show that G protein-coupled receptor class C group 5 member B(GPRC5B),an orphan G-protein-couple receptor,not only inhibits cartilage degradation,but also increases cartilage regeneration and thereby is protective against OA.We observed that Gprc5b deficient chondrocytes had an upregulation of cartilage catabolic gene expression,along with downregulation of anabolic genes in vitro.Furthermore,mice deficient in Gprc5b displayed a more severe OA phenotype in the destabilization of the medial meniscus(DMM)induced OA mouse model,with upregulation of cartilage catabolic factors and downregulation of anabolic factors,consistent with our in vitro findings.Overexpression of Gprc5b by lentiviral vectors alleviated the cartilage degeneration in DMM-induced OA mouse model by inhibiting cartilage degradation and promoting regeneration.We also assessed the molecular mechanisms downstream of Gprc5b that may mediate these observed effects and identify the role of protein kinase B(AKT)-mammalian target of rapamycin(m TOR)-autophagy signaling pathway.Thus,we demonstrate an integral role of GPRC5B in OA pathogenesis,and activation of GPRC5B has the potential in preventing the progression of OA.展开更多
Organic anion transporting polypeptide 1 B1 and 1 B3(OATP1 B1/3)as important uptake transporters play a fundamental role in the transportation of exogenous drugs and endogenous substances into cells.Rat OATP1 B2,encod...Organic anion transporting polypeptide 1 B1 and 1 B3(OATP1 B1/3)as important uptake transporters play a fundamental role in the transportation of exogenous drugs and endogenous substances into cells.Rat OATP1 B2,encoded by the Slcolb2 gene,is homologous to human OATP1 B1/3.Although OATP1 B1/3 is very important,few animal models can be used to study its properties.In this report,we successfully constructed the Slco1 b2 knockout(KO)rat model via using the CRISPR/Cas9 technology for the first time.The novel rat model showed the absence of OATP1 B2 protein expression,with no offtarget effects as well as compensatory regulation of other transporters.Further pharmacokinetic study of pitavastatin,a typical substrate of OATP1 B2,confirmed the OATP1 B2 function was absent.Since bilirubin and bile acids are the substrates of OATP1 B2,the contents of total bilirubin,direct bilirubin,indirect bilirubin,and total bile acids in serum are significantly higher in Slco1 b2 KO rats than the data of wild-type rats.These results are consistent with the symptoms caused by the absence of OATP1 B1/3 in Rotor syndrome.Therefore,this rat model is not only a powerful tool for the study of OATP1 B2-mediated drug transportation,but also a good disease model to study hyperbilirubinemia-related diseases.展开更多
The laboratory rat is a valuable mammalian model organism for basic research and drug discovery. Here we demonstrate an efficient methodology by applying transcription activator-like effector nucleases(TALENs) technol...The laboratory rat is a valuable mammalian model organism for basic research and drug discovery. Here we demonstrate an efficient methodology by applying transcription activator-like effector nucleases(TALENs) technology to generate Leptin receptor(Lepr) knockout rats on the Sprague Dawley(SD) genetic background. Through direct injection of in vitro transcribed m RNA of TALEN pairs into SD rat zygotes, somatic mutations were induced in two of three resulting pups. One of the founders carrying bi-allelic mutation exhibited early onset of obesity and infertility. The other founder carried a chimeric mutation which was efficiently transmitted to the progenies. Through phenotyping of the resulting three lines of rats bearing distinct mutations in the Lepr locus, we found that the strains with a frame-shifted or premature stop codon mutation led to obesity and metabolic disorders. However, no obvious defect was observed in a strain with an in-frame 57 bp deletion in the extracellular domain of Lepr. This suggests the deleted amino acids do not significantly affect Lepr structure and function. This is the first report of generating the Lepr mutant obese rat model in SD strain through a reverse genetic approach. This suggests that TALEN is an efficient and powerful gene editing technology for the generation of disease models.展开更多
Among the most important sensors of extracellular danger signals, purinergic receptors have been demonstrated to play crucial roles in host defense against infection. However, the function of P2 receptors in viral inf...Among the most important sensors of extracellular danger signals, purinergic receptors have been demonstrated to play crucial roles in host defense against infection. However, the function of P2 receptors in viral infection has been little explored. Here we demonstrated that P2Y13 and its ligand ADP play an important role in protecting hosts from viral infections. First, we demonstrate that P2Y13, as a typical interferon-stimulated gene, is induced together with extracellular ADP during viral infection. Most importantly, extracellular ADP restricts the replication of different kinds of viruses, including vesicular stomatitis virus, Newcastle disease virus, herpes simplex virus 1, and murine leukemia virus. This kind of protection is dependent on P2Y13 but not P2Y1 or P2Y12, which are also considered as receptors for ADP. Furthermore, cyclic adenosine monophosphate and EPAC1 are downregu-lated by extracellular ADP through the P2Y13-coupled Gi alpha subunit. Accordingly, inhibition or deletion of EPAC1 significantly eliminates ADP/P2Y13-mediated antiviral activities. Taken together, our results show that P2Y13 and ADP play pivotal roles in the clearance of invaded virus and have the potential as antiviral targets.展开更多
As the most prominent clinical drug targets for the inhibition of platelet aggregation, P2Y12 and P2Y13 have been found to be highly expressed in both platelets and macrophages. However, the roles and function of P2Y1...As the most prominent clinical drug targets for the inhibition of platelet aggregation, P2Y12 and P2Y13 have been found to be highly expressed in both platelets and macrophages. However, the roles and function of P2Y12/13 in the regulation of macrophage-mediated innate immune responses remain unclear. Here, we demonstrate that adenosine 5′-diphosphate (ADP), the endogenous ligand of P2Y1, P2Y12 and P2Y13, was released both in E. coli-infected mice and from macrophages treated with either lipopolysaccharide (LPS) or Pam3CSK4. Furthermore, the expression of P2Y13 was clearly increased in both LPS-treated macrophages and tuberculosis patients. ADP protected mice from E. coli 0111-induced peritonitis by recruiting more macrophages to the infected sites. Consistent with this, ADP and ADP-treated cell culture medium attracted more macrophages in the transwell assay by enhancing the expression of MCP-1. Nevertheless, P2Y1 is dispensable for ADP-mediated protection against bacterial infection. However, either P2Y12/P2Y13 deficiency or blocking the downstream signaling of P2Y12/P2Y13 blocked the ADP-mediated immune response and allowed more bacteria to persist in the infected mice. Furthermore, extracellular signal-regulated kinase (ERK) phosphorylation was clearly increased by ADP, and this type of activation could be blocked by either forskolin or analogs of cyclic AMP (cAMP) (for example, 8-bromo-cAMP). Accordingly, ADP-induced MCP-1 production and protection against bacterial infection could also be reduced by U0126, forskolin and 8-bromo-cAMP. Overall, our study reveals a relationship between danger signals and innate immune responses, which suggests the potential therapeutic significance of ADP-mediated purinergic signaling in infectious diseases.展开更多
Cholesterol is an important precursor of many endogenous molecules.Disruption of cholesterol homeostasis can cause many pathological changes,leading to liver and cardiovascular diseases.CYP1A is widely involved in cho...Cholesterol is an important precursor of many endogenous molecules.Disruption of cholesterol homeostasis can cause many pathological changes,leading to liver and cardiovascular diseases.CYP1A is widely involved in cholesterol metabolic network,but its exact function has not been fully elucidated.Here,we aim to explore how CYP1A regulates cholesterol homeostasis.Our data showed that CYP1A1/2 knockout(KO)rats presented cholesterol deposition in blood and liver.The serum levels of low-density lipoprotein cholesterol,high-density lipoprotein cholesterol and total cholesterol were significantly increased in KO rats.Further studies found that the lipogenesis pathway(LXRa-SREBP1-SCD1)of KO rats was activated,and the key protein of cholesterol ester hydrolysis(CES1)was inhibited.Importantly,lansoprazole can significantly alleviate rat hepatic lipid deposition in hypercholesterolemia models by inducing CYP1A.Our findings reveal the role of CYP1A as a potential regulator of cholesterol homeostasis and provide a new perspective for the treatment of hypercholesterolemia.展开更多
基金National Key Research and Development Program of China(2023YFB3810200 to J.L.)National Natural Science Foundation of China(92168204,82225030 to J.L.)Fundamental Research Funds for the Central Universities(22120210586 to J.L.)。
文摘Reproductive hormones associated with the hypothalamic-pituitary-gonadal(HPG)axis are closely linked to bone homeostasis.In this study,we demonstrate that Gonadotropin inhibitory hormone(GnIH,one of the key reproductive hormones upstream of the HPG axis)plays an indispensable role in regulating bone homeostasis and maintaining bone mass.We find that deficiency of GnIH or its receptor Gpr147 leads to a significant reduction in bone mineral density(BMD)in mice primarily by enhancement of osteoclast activation in vivo and in vitro.Mechanistically,GnIH/Gpr147 inhibits osteoclastogenesis by the PI3K/AKT,MAPK,NF-κB and Nfatc1 signaling pathways.Furthermore,GnIH treatment was able to alleviate bone loss in aging,ovariectomy(OVX)or LPS-induced mice.Moreover,the therapy using green light promotes the release of GnIH and rescues OVX-induced bone loss.In humans,serum GnIH increases and bone resorption markers decrease after green light exposure.Therefore,our study elucidates that GnIH plays an important role in maintaining bone homeostasis via modulating osteoclast differentiation and demonstrates the potential of GnIH therapy or green light therapy in preventing osteoporosis.
基金supported by grants from the National Key Research and Development Program of China (2020YFC2002800 to J.L. and 2018YFC1105102 to J.L.)the National Natural Science Foundation of China (91949127, 92168204 to J.L.)the Fundamental Research Funds for the Central Universities (22120210586)
文摘Prostaglandin E2(PGE2), a major cyclooxygenase-2(COX-2) product, is highly secreted by the osteoblast lineage in the subchondral bone tissue of osteoarthritis(OA) patients. However, NSAIDs, including COX-2 inhibitors, have severe side effects during OA treatment. Therefore, the identification of novel drug targets of PGE2 signaling in OA progression is urgently needed. Osteoclasts play a critical role in subchondral bone homeostasis and OA-related pain. However, the mechanisms by which PGE2 regulates osteoclast function and subsequently subchondral bone homeostasis are largely unknown. Here, we show that PGE2 acts via EP4 receptors on osteoclasts during the progression of OA and OA-related pain. Our data show that while PGE2 mediates migration and osteoclastogenesis via its EP2 and EP4 receptors, tissue-specific knockout of only the EP4 receptor in osteoclasts(EP4 Lys M) reduced disease progression and osteophyte formation in a murine model of OA. Furthermore, OA-related pain was alleviated in the EP4 Lys M mice, with reduced Netrin-1 secretion and CGRP-positive sensory innervation of the subchondral bone. The expression of plateletderived growth factor-BB(PDGF-BB) was also lower in the EP4 Lys Mmice, which resulted in reduced type H blood vessel formation in subchondral bone. Importantly, we identified a novel potent EP4 antagonist, HL-43, which showed in vitro and in vivo effects consistent with those observed in the EP4 Lys Mmice. Finally, we showed that the Gαs/PI3 K/AKT/MAPK signaling pathway is downstream of EP4 activation via PGE2 in osteoclasts. Together, our data demonstrate that PGE2/EP4 signaling in osteoclasts mediates angiogenesis and sensory neuron innervation in subchondral bone, promoting OA progression and pain, and that inhibition of EP4 with HL-43 has therapeutic potential in OA.
基金supported by grants from the National Key Research and Development Program of China(2018YFC1105102 to J.L.,2016YFC0902102 to J.L.and J.X.)the National Natural Science Foundation of China(81722020,91749204,81472048 to J.L.,81330049 to M.L.,81330059 and 81572640 to J.X.)+2 种基金the Innovation Program of Shanghai Municipal Education Commission(14ZZ051 to J.L.,2017ZZ01017 to J.X.)the Science and Technology Commission of Shanghai Municipality(12ZR1447900 to J.L.,17JC1400903 and 17411950300 to J.X.)the Fundamental Research Funds for the Central Universities(to J.L.)
文摘The superfamily of G protein-coupled receptors (GPCRs) contains immense structural and functional diversity and mediates a myriad of biological processes upon activation by various extracellular signals.Critical roles of GPCRs have been established in bone development,remodeling,and disease.Multiple human GPCR mutations impair bone development or metabolism,resulting in osteopathologies.Here we summarize the disease phenotypes and dysfunctions caused by GPCR gene mutations in humans as well as by deletion in animals.To date,92 receptors (5 glutamate family,67 rhodopsin family,5 adhesion,4 frizzled/taste2 family,5 secretin family,and 6 other 7TM receptors) have been associated with bone diseases and dysfunctions (36 in humans and 72 in animals).By analyzing data from these 92 GPCRs,we found that mutation or deletion of different individual GPCRs could induce similar bone diseases or dysfunctions,and the same individual GPCR mutation or deletion could induce different bone diseases or dysfunctions in different populations or animal models.Data from human diseases or dysfunctions identified 19 genes whose mutation was associated with human BMD:9 genes each for human height and osteoporosis;4 genes each for human osteoarthritis (OA) and fracture risk;and 2 genes each for adolescent idiopathic scoliosis (AIS),periodontitis,osteosarcoma growth,and tooth development.Reports from gene knockout animals found 40 GPCRs whose deficiency reduced bone mass,while deficiency of 22 GPCRs increased bone mass and BMD;deficiency of 8 GPCRs reduced body length,while 5 mice had reduced femur size upon GPCR deletion.Furthermore,deficiency in 6 GPCRs induced osteoporosis;4 induced osteoarthritis;3 delayed fracture healing;3 reduced arthritis severity;and reduced bone strength,increased bone strength,and increased cortical thickness were each observed in 2 GPCR-deficiency models.The ever-expanding number of GPCR mutation-associated diseases warrants accelerated molecular analysis,population studies,and investigation of phenotype correlation with SNPs to elucidate GPCR function in human diseases.
基金Supported by Grants-in-Aid for Young Scientists(B)No.22791295,No.23791557 and No.24791440Grant-in-Aid for Scientific Research(C)from the Japan Society for the Promotion of Science,No.22591464 and No.24591957
文摘AIM: To investigate the role of claudin 1 in the regulation of genes involved in cell migration and tumor necrosis factor alpha (TNF-α)-induced gene expression in human gastric adenocarcinoma cells.
基金partially supported by Major State Basic Research Development Program of China(Nos.2015CB910400 and 2012CB910400)National Natural Science Foundation of China(Nos.81472788 and 81272463)
文摘Prostate cancer (PCa) is the most common male cancer [1, 2]. PCa initially depends on androgen receptor (AR) signaling for growth and survival. Androgen deprivation therapy causes a temporary reduction in PCa tumor burden, but the tumor eventually develops into castrationresistant prostate cancer (CRPC) with the ability to grow again in the absence of androgens [3]. Mechanisms of CRPC progression include AR amplification and overexpression [4], AR gene rearrangement promoting synthesis of constitutively-active truncated AR splice variants (ARVs) [4], and induction of intracrine androgen metabolic enzymes [3]. Current anti-androgen therapies including MDV3100 (Enzalutamide) and abiraterone have focused on the androgen-dependent activation of AR through its ligand-binding domain (LBD), but do not provide a continuing clinical benefit for patients with CRPC and presumably fail due to multiple mechanisms including the expression of AR-Vs lacking the LBD [5]. These AR-Vs signal in the absence of ligand and are therefore resistant to LBD-targeting AR antagonists or agents that repress androgen biosynthesis [6].
基金supported by grants from Canadian Institutes of Health Research(Grant Nos.:MOP-42546,MOP-119514,PJT148847).
文摘Use of gold nanoparticles(GNPs)in medicine is an emerging field of translational research with vast clinical implications and exciting therapeutic potential.However,the safety of using GNPs in human subjects is an important question that remains unanswered.This study reviews over 20 clinical trials focused on GNP safety and aims to summarize all the clinical studies,completed and ongoing,to identify whether GNPs are safe to use in humans as a therapeutic platform.In these studies,GNPs were implemented as drug delivery devices,for photothermal therapy,and utilized for their intrinsic therapeutic effects by various routes of delivery.These studies revealed no major safety concerns with the use of GNPs;however,the number of trials and total patient number remains limited.Multi-dose,multicenter blinded trials are required to deepen our understanding of the use of GNPs in clinical settings to facilitate translation of this novel,multifaceted therapeutic device.Expanding clinical trials will require collaboration between clinicians,scientists,and biotechnology companies.
基金supported by grants from National Key R&D Program of China(2019YFA0110802 and 2019YFA0802800)the National Natural Science Foundation of China(32025023,31971366)+1 种基金grants from the Shanghai Municipal Commission for Science and Technology(21CJ1402200,20140900200)the Innovation Program of Shanghai Municipal Education Commission(2019-01-07-00-05-E00054)。
文摘CRISPR/Cas9-mediated site-specific insertion of exogenous genes holds potential for clinical applications.However,it is still infeasible because homologous recombination(HR)is inefficient,especially for nondividing cells.To overcome the challenge,we report that a homology-independent targeted integration(HITI)strategy is used for permanent integration of high-specificity-activity Factor IX variant(F9 Padua,R338L)at the albumin(Alb)locus in a novel hemophilia B(HB)rat model.The knock-in efficiency reaches 3.66%,as determined by droplet digital PCR(dd PCR).The clotting time is reduced to a normal level four weeks after treatment,and the circulating factor IX(FIX)level is gradually increased up to 52%of the normal level over nine months even after partial hepatectomy,demonstrating the amelioration of hemophilia.Through primer-extension-mediated sequencing(PEM-seq),no significant off-target effect is detected.This study not only provides a novel model for HB but also identifies a promising therapeutic approach for rare inherited diseases.
基金supported by grants from the National Natural Science Foundation of China(Grant 81672758)the Science and Technology Commission of Shanghai Municipality(Grants 16ZR1410400 and 14DZ2270100)the Fundamental Research Funds for the Central Universities(Grant 78260029)
文摘The KRAS gene is frequently mutated in multiple cancer types,but it fell off the drug discovery radar for many years because of its inherent "undruggable" structure and undefined biological properties.As reported in the paper entitled "Suppression of KRas-mutant cancer through the combined inhibition of KRAS with PLK1 and ROCK" in Nature Communications,we performed a synthetic lethal screening with a combinatorial strategy on a panel of clinical drugs;we found that combined inhibition of polo-like kinase 1 and RhoA/Rho kinase markedly suppressed tumor growth in mice.An increase in the expression of the tumor suppressor P21^(WAF1/CIP1) contributed to the synergistic mechanism of the combination therapy.These findings open a novel avenue for the treatment of KRAS-mutant lung cancer.
基金financially supported by the National Natural Science Foundation of China(grant no.21573230)973 National Basic Research Program of the Ministry of Science and Technology(grant no.2014CB239400)
文摘Energy storage is pivotal for the continuous utilization of solar energy suffering from the intermittency issue. Herein, we demonstrate a solar rechargeable flow cell（SRFC） based on photoelectrochemical regeneration of vanadium redox species for in-situ solar energy harvest and storage. In this device, TiO_2 and MWCNT/acetylene black（MWCNT/AB） composite are served as the photoanode and the counter electrode,respectively, with all vanadium redox couples, VO_2~＋/VO^（2＋）and VO^（2＋）/V^（3＋）, as solar energy storage media.Benefitting from solar energy, the cell can be photocharged under a bias as low as 0.1 V, which is much lower than the discharge voltage of ~0.5 V. Photocharged under the optimized condition, the cell delivers a discharge energy of 23.0 mWh/L with 67.4% input electric energy savings. This prototype work may inspire the rational design for cost-effective solar energy storage devices.
基金National Key R&D Program of China(2019YFA0110802 and 2019YFA0802800)the National Natural Science Foundation of China(32025023,31971366)+1 种基金the Shanghai Municipal Commission for Science and Technology(21JC1402200,20140900200)the Innovation Program of Shanghai Municipal Education Commission(2019-01-07-00-05-E00054)。
文摘The authors regret that the grant number“21CJ1402200”in the Acknowledgments session should be replaced as“21JC1402200”.The corrected contents areprovided as follows.
基金supported by the grants from the National Natural Science Foundation of China(No.82073310,82373146,82202897)The Jointed PI Program from Shanghai Changning Maternity and Infant Health Hospital(China)(No.PI202430)+3 种基金Shanghai Rising-Star Program(No.23QB1405600)The Science and Technology Commission of Shanghai Municipality,China(No.22QB1405600)Biomedical Projects of Yunnan Key Science and Technology Program(202502AA310002)ECNU(East China Normal University)Construction Fund of Innovation and Entrepreneurship Laboratory(Shanghai,China)(No.44400-20201-532300/021).
文摘Glioblastoma multiforme(GBM)is the deadliest form of brain tumor,and effective treatments are lacking.Thus,a new generation of effective treatments is urgently needed.B-cell lymphoma 6(BCL6)is a transcription factor that functions to suppress the transcription of DNA damage response genes,halting cell death in response to DNA damage.Here,we identified BCL6 as a lynchpin in GBM,the expression of which was greater in GBM cells than in normal cells and associated with poor survival in GBM patients.The silencing of BCL6 additionally affected GBM cell proliferation and triggered cellular damage.Furthermore,we reported the identification of YK01,a novel small-molecule inhibitor of BCL6.YK01 exhibited excellent anti-GBM bioactivity and caused apoptosis;importantly,YK01 significantly inhibited the growth of GBM cells both in vitro and in vivo.Moreover,the combination of YK01 and temozolomide treatment significantly suppressed the growth and metastasis of tumors in vivo and prolonged the survival of mice with tumors.In summary,our findings reveal that BCL6 appears to play a crucial role in GBM and may be a therapeutic target for treating this incurable condition.
基金supported by the National Natural Science Foundation of China (62425204, 62122025, U22A2037, 62450002,62432011, 62250028, 81972828, 82172644, and 81830083)Hunan Provincial Natural Science Foundation of China(2021JJ10020)+1 种基金National Key Scientific Infrastructure for Translational Medicine (Shanghai)(TMSK-2021-120)ECNU Multifunctional Platform for Innovation (011)
文摘In target-based drug design,the manual creation of a poor initial compound library,the time-consuming wetlaboratory experimental screening method,and the weak explainability of their activity against compounds significantly limit the efficiency of discovering novel therapeutics.Here we propose an image-guided,interpretability deep learning workflow,named LeadDisFlow,to enable rapid,accurate target drug discovery.Using LeadDisFlow,we identified four potent antagonists with single-nanomolar antagonistic activity against PGE2 receptor subtype 4(EP4),a promising target for tumor im-munotherapy.Remarkably,the most potent EP4 antagonist,ZY001,demonstrated an IC50 value of(0.51±0.02)nM,along with high selectivity.Furthermore,ZY001 effectively impaired the PGE2-induced gene expression of a panel of immunosuppressive molecules in macrophages.The workflow facilitates the discovery of potent EP4 antagonists that enhance anti-tumor immune response,and provides a convenient and quick approach to discover promising therapeutics for a specific drug target.
基金supported by the National Natural Science Foundation of China (31371455, 31171318 to Dali Li, 81330049 to Mingyao Liu)the Science and Technology Commission of Shanghai Municipality (14140900300 to Dali Li)
文摘CRISPR/Cas9 is a versatile genome-editing tool which is widely used for modifying the genome of both prokaryotic and eukaryotic organisms for basic research and applications. An increasing number of reports have demonstrated that CRISPR/Cas9-mediated genome editing is a powerful technology for gene therapy. Here, we review the recent advances in CRISPR/Cas9-mediated gene therapy in animal models via different strategies and discuss the challenges as well as future prospects.
基金supported by grants from the National Key Research and Development Program of China(2020YFC2002800 and 2018YFC1105102 to Jian Luo)the National Natural Science Foundation of China(82225030,92168204 and 9194910271 to Jian Luo)+2 种基金Shanghai Municipal Health Commission Excellent Young Medical Talents Training Program(2022XD034 to Jian Luo,China)the Fundamental Research Funds for the Central Universities(22120210586 to Jian Luo,China)the East China Normal University(ECNU)Multifunctional Platform for Innovation(011)。
文摘Osteoarthritis(OA)is one of the most common chronic diseases in the world.However,current treatment modalities mainly relieve pain and inhibit cartilage degradation,but do not promote cartilage regeneration.In this study,we show that G protein-coupled receptor class C group 5 member B(GPRC5B),an orphan G-protein-couple receptor,not only inhibits cartilage degradation,but also increases cartilage regeneration and thereby is protective against OA.We observed that Gprc5b deficient chondrocytes had an upregulation of cartilage catabolic gene expression,along with downregulation of anabolic genes in vitro.Furthermore,mice deficient in Gprc5b displayed a more severe OA phenotype in the destabilization of the medial meniscus(DMM)induced OA mouse model,with upregulation of cartilage catabolic factors and downregulation of anabolic factors,consistent with our in vitro findings.Overexpression of Gprc5b by lentiviral vectors alleviated the cartilage degeneration in DMM-induced OA mouse model by inhibiting cartilage degradation and promoting regeneration.We also assessed the molecular mechanisms downstream of Gprc5b that may mediate these observed effects and identify the role of protein kinase B(AKT)-mammalian target of rapamycin(m TOR)-autophagy signaling pathway.Thus,we demonstrate an integral role of GPRC5B in OA pathogenesis,and activation of GPRC5B has the potential in preventing the progression of OA.
基金supported in whole or part by grants from the National Natural Science Foundation of China(No.81773808)the Science and Technology Commission of Shanghai Municipality(Nos.17140901000,17140901001 and 18430760400)supported from ECNU Multifunctional Platform for Innovation(011,China)
文摘Organic anion transporting polypeptide 1 B1 and 1 B3(OATP1 B1/3)as important uptake transporters play a fundamental role in the transportation of exogenous drugs and endogenous substances into cells.Rat OATP1 B2,encoded by the Slcolb2 gene,is homologous to human OATP1 B1/3.Although OATP1 B1/3 is very important,few animal models can be used to study its properties.In this report,we successfully constructed the Slco1 b2 knockout(KO)rat model via using the CRISPR/Cas9 technology for the first time.The novel rat model showed the absence of OATP1 B2 protein expression,with no offtarget effects as well as compensatory regulation of other transporters.Further pharmacokinetic study of pitavastatin,a typical substrate of OATP1 B2,confirmed the OATP1 B2 function was absent.Since bilirubin and bile acids are the substrates of OATP1 B2,the contents of total bilirubin,direct bilirubin,indirect bilirubin,and total bile acids in serum are significantly higher in Slco1 b2 KO rats than the data of wild-type rats.These results are consistent with the symptoms caused by the absence of OATP1 B1/3 in Rotor syndrome.Therefore,this rat model is not only a powerful tool for the study of OATP1 B2-mediated drug transportation,but also a good disease model to study hyperbilirubinemia-related diseases.
基金supported by the State Key Development Programs of China (2012CB910400 to Mingyao Liu)the National Natural Science Foundation of China (31371455, 31171318 and 81330049)the Science and Technology Commission of Shanghai Municipality (14140900300)
文摘The laboratory rat is a valuable mammalian model organism for basic research and drug discovery. Here we demonstrate an efficient methodology by applying transcription activator-like effector nucleases(TALENs) technology to generate Leptin receptor(Lepr) knockout rats on the Sprague Dawley(SD) genetic background. Through direct injection of in vitro transcribed m RNA of TALEN pairs into SD rat zygotes, somatic mutations were induced in two of three resulting pups. One of the founders carrying bi-allelic mutation exhibited early onset of obesity and infertility. The other founder carried a chimeric mutation which was efficiently transmitted to the progenies. Through phenotyping of the resulting three lines of rats bearing distinct mutations in the Lepr locus, we found that the strains with a frame-shifted or premature stop codon mutation led to obesity and metabolic disorders. However, no obvious defect was observed in a strain with an in-frame 57 bp deletion in the extracellular domain of Lepr. This suggests the deleted amino acids do not significantly affect Lepr structure and function. This is the first report of generating the Lepr mutant obese rat model in SD strain through a reverse genetic approach. This suggests that TALEN is an efficient and powerful gene editing technology for the generation of disease models.
基金the National Key R&D Program of China (2018YFA0507000 to B.D.)the National Natural Science Foundation of China (31570896 and 31770969 to B.D., 81672811 to M.Q.)+2 种基金Joint Research Institute for Science and Society (JoRISS)(14JORISS01 to B.D.)Science and Technology Commission of Shanghai Municipality (15JC14O15OO to B.D.)Innovation Program of Shanghai Municipal Educati on Commissi on (2017-01- 07-00-05-E00011 to A/LL.).
文摘Among the most important sensors of extracellular danger signals, purinergic receptors have been demonstrated to play crucial roles in host defense against infection. However, the function of P2 receptors in viral infection has been little explored. Here we demonstrated that P2Y13 and its ligand ADP play an important role in protecting hosts from viral infections. First, we demonstrate that P2Y13, as a typical interferon-stimulated gene, is induced together with extracellular ADP during viral infection. Most importantly, extracellular ADP restricts the replication of different kinds of viruses, including vesicular stomatitis virus, Newcastle disease virus, herpes simplex virus 1, and murine leukemia virus. This kind of protection is dependent on P2Y13 but not P2Y1 or P2Y12, which are also considered as receptors for ADP. Furthermore, cyclic adenosine monophosphate and EPAC1 are downregu-lated by extracellular ADP through the P2Y13-coupled Gi alpha subunit. Accordingly, inhibition or deletion of EPAC1 significantly eliminates ADP/P2Y13-mediated antiviral activities. Taken together, our results show that P2Y13 and ADP play pivotal roles in the clearance of invaded virus and have the potential as antiviral targets.
基金This work was supported by the National Basic Research Program of China(2012CB910400)the National Natural Science Foundation of China(81272369,31470040,31570896 and 81672811)+1 种基金the Doctoral Fund of Ministry of Education of China(20130076110013)Science and Technology Commission of Shanghai Municipality(15JC1401500).
文摘As the most prominent clinical drug targets for the inhibition of platelet aggregation, P2Y12 and P2Y13 have been found to be highly expressed in both platelets and macrophages. However, the roles and function of P2Y12/13 in the regulation of macrophage-mediated innate immune responses remain unclear. Here, we demonstrate that adenosine 5′-diphosphate (ADP), the endogenous ligand of P2Y1, P2Y12 and P2Y13, was released both in E. coli-infected mice and from macrophages treated with either lipopolysaccharide (LPS) or Pam3CSK4. Furthermore, the expression of P2Y13 was clearly increased in both LPS-treated macrophages and tuberculosis patients. ADP protected mice from E. coli 0111-induced peritonitis by recruiting more macrophages to the infected sites. Consistent with this, ADP and ADP-treated cell culture medium attracted more macrophages in the transwell assay by enhancing the expression of MCP-1. Nevertheless, P2Y1 is dispensable for ADP-mediated protection against bacterial infection. However, either P2Y12/P2Y13 deficiency or blocking the downstream signaling of P2Y12/P2Y13 blocked the ADP-mediated immune response and allowed more bacteria to persist in the infected mice. Furthermore, extracellular signal-regulated kinase (ERK) phosphorylation was clearly increased by ADP, and this type of activation could be blocked by either forskolin or analogs of cyclic AMP (cAMP) (for example, 8-bromo-cAMP). Accordingly, ADP-induced MCP-1 production and protection against bacterial infection could also be reduced by U0126, forskolin and 8-bromo-cAMP. Overall, our study reveals a relationship between danger signals and innate immune responses, which suggests the potential therapeutic significance of ADP-mediated purinergic signaling in infectious diseases.
基金supported in whole or part by grants from the National Natural Science Foundation of China(81773808,82274010)the Science and Technology Commission of Shanghai Municipality(18430760400,China)+4 种基金the Jointed PI Program from Shanghai Changning Maternity and Infant Health Hospital(2019CNECNUPI02,China)the Fundamental Research Funds for the Central Universities(China)ECNU Construction Fund of Innovation and Entrepreneurship Laboratory(China)supported from ECNU Multifunctional Platform for Innovation(011,China)the Instruments Sharing Platform of School of Life Sciences,East China Normal University(Shanghai,China)。
文摘Cholesterol is an important precursor of many endogenous molecules.Disruption of cholesterol homeostasis can cause many pathological changes,leading to liver and cardiovascular diseases.CYP1A is widely involved in cholesterol metabolic network,but its exact function has not been fully elucidated.Here,we aim to explore how CYP1A regulates cholesterol homeostasis.Our data showed that CYP1A1/2 knockout(KO)rats presented cholesterol deposition in blood and liver.The serum levels of low-density lipoprotein cholesterol,high-density lipoprotein cholesterol and total cholesterol were significantly increased in KO rats.Further studies found that the lipogenesis pathway(LXRa-SREBP1-SCD1)of KO rats was activated,and the key protein of cholesterol ester hydrolysis(CES1)was inhibited.Importantly,lansoprazole can significantly alleviate rat hepatic lipid deposition in hypercholesterolemia models by inducing CYP1A.Our findings reveal the role of CYP1A as a potential regulator of cholesterol homeostasis and provide a new perspective for the treatment of hypercholesterolemia.
