关键词： Asporin: Cd109   Spinal cord injury   snRNA-seq   Axonal regeneration   TGF-beta 1  

摘要： Axonal regeneration is restricted in adults and causes irreversible motor dysfunction following spinal cord injury (SCI). In contrast, neonates have prominent regenerative potential and can restore their neural function. Although the distinct cellular responses in neonates have been studied, how they contribute to neural recovery remains unclear. To assess whether the secreted molecules in neonatal SCI can enhance neural regeneration, we re-analyzed the previously performed single-nucleus RNA-seq (snRNA-seq) and focused on Asporin and Cd109, the highly expressed genes in the injured neonatal spinal cord. In the present study, we showed that both these molecules were expressed in the injured spinal cords of adults and neonates. We treated the cortical neurons with recombinant Asporin or CD109 to observe their direct effects on neurons in vitro. We demonstrated that these molecules enhance neurite outgrowth in neurons. However, these molecules did not enhance re-growth of severed axons. Our results suggest that Asporin and CD109 influence neurites at the lesion site, rather than promoting axon regeneration, to restore neural function in neonates after SCI.

关键词： Asporin   PLAP-1   SLRPs   TGF beta   ECM   EMT  

摘要： Asporin (ASPN) has been identified as one of the members of the class I small leucine-rich proteoglycans (SLRPs) family in the extracellular matrix (ECM). It is involved in classic ensigns of cancers such as self-dependent growth, resistance to growth inhibitors, restricting apoptosis, cancer metastasis, and bone-related disorders. ASPN is different from other members of SLRPs, such as decorin (DCN) and biglycan (BGN), in a way that it contains a distinctive length of aspartate (D) residues in the amino (N) -terminal region. These D-repeats residues possess germline polymorphisms and are identified to be linked with cancer progression and osteoarthritis (OA). The polyaspartate stretch in the N-terminal region of the protein and its resemblance to DCN are the reasons it is called asporin. In this review, we comprehensively summarized and updated the dual role of ASPN in various malignancies, its structure in mice and humans, variants, mutations, cancer-associated signalings and functions, the relationship between ASPN and cancer-epithelial, stromal fibroblast crosstalk, immune cells and immunosuppression in cancer and other diseases. In cancer and other bone-related diseases, ASPN is identified to be regulating various signaling pathways such as TGF beta, Wnt/beta-catenin, notch, hedgehog, EGFR, HER2, and CD44mediated Rac1. These pathways promote cancer cell invasion, proliferation, and migration by mediating the epithelial-to-mesenchymal transition (EMT) process. Finally, we discussed mouse models mimicking ASPN in vivo function in cancers and the probability of therapeutic targeting of ASPN in cancer cells, fibrosis, and other bonerelated diseases.

关键词： Pancancer   Bladder cancer   Molecular biomarker   Prognosis   Immune   Cancer therapy  

摘要： Background: Asporin (ASPN) has been identified as a player in tumorigenesis, but its precise roles and modulatory function are largely unknown. Methods: In the present study, ASPN expression was first explored, followed by a prognostic evaluation of ASPN and a comprehensive investigation of the connections between ASPN and immunomodulation, immune cell infiltration and potential compounds on a pancancer level. Finally, ASPN expression was validated in bladder urothelial carcinoma (BLCA) tissues, and the potential function of ASPN, including its effects on migration and invasion capabilities, was investigated in tumor cells. Results: The expression of ASPN exhibited significant variation across cancers and was found to be associated with patient prognosis. In addition, the expression level of APSN was markedly correlated with the abundances of infiltrating immune cells and cancer-associated fibroblasts and the expression levels of immunomodulatory genes based on the results of pancancer analysis. Metastasis and immune-associated signaling pathways were identified in enrichment analysis based on ASPN expression. Finally, we confirmed that ASPN expression increased with the degree of malignancy in BLCA tissues and cell lines and that low expression of ASPN hindered the migration and invasion of cells. Conclusions: ASPN has the potential to be a biomarker of cancer prognosis and a therapeutic target, and it also has predictive capability for the progression of BLCA.

关键词： 食管鳞状细胞癌   无孢蛋白   癌旁正常组织   癌基因  

摘要： 目的探讨无孢蛋白(Asporin)在食管鳞状细胞癌发生发展中的作用及临床意义。方法收集2019年12月⁃2021年11月选取涟水人民医院收治的140例食管鳞状细胞癌患者。随机选取60例新鲜切取的食管鳞状细胞癌(ESCC)组织及癌旁正常组织,利用Western blot、RT⁃PCR技术检测Asporin的蛋白及mRNA表达水平;80例ESCC组织及其相应癌旁组织的石蜡标本,利用免疫组化技术进一步检测Asporin的表达情况;利用卡方检验、Kaplan⁃Meier分析以及COX回归分析验证Asporin的表达与ESCC患者临床病理特征及预后的关系;Transwell侵袭实验研究Asporin沉默后对ESCC细胞株侵袭转移功能的影响;Pearson相关分析评价Asporin与基质金属蛋白酶2(MMP2)之间的相关性。结果Western blot和RT⁃PCR结果显示ESCC的Asporin蛋白和mRNA表达水平明显高于其癌旁正常组织(P<0.05);免疫组化结果显示,Asporin在ESCC组织中的表达明显高于癌旁组织,且Asporin在ESCC的高表达率为65%(52/80);Asporin表达水平与患者的淋巴结转移情况及TNM分期有显著相关性(P<0.05),与其他临床病理特征无显著相关性(P>0.05);Kaplan⁃Meier分析结果表明,高表达Asporin的ESCC患者无病生存期短于低表达ASPN的患者(P<0.01);COX回归分析显示,低表达Asporin的ESCC患者死亡风险低于高表达ASPN的患者,HR=0.106(95%CI:0.024⁃0.469),Asporin是影响ESCC患者预后的独立危险因素(P=0.003);Transwell侵袭实验表明,经siRNA沉默Asporin后,与阴性对照组相比,Asporin敲低组的侵袭能力明显降低;Pearson相关分析显示,Asporin的表达水平与MMP2表达存在中度正相关关系(r=0.61,P<0.01),MMP2在恶性肿瘤侵袭转移过程中起关键作用。结论Asporin可能作为一种癌基因,参与食管鳞状细胞癌的发生发展。

关键词： Asporin   Osteocalcin   Osteopontin   Osteoblast   BMP2   Runx2   RNA-Seq   Anti-miR   Fetuin A   Differentiation  

摘要： BackgroundCalcific aortic valve disease (CAVD) is the second leading cause of adult heart diseases. The purpose of this study is to investigate whether miR-101-3p plays a role in the human aortic valve interstitial cells (HAVICs) calcification and the underlying *** RNA deep sequencing and qPCR analysis were used to determine changes in microRNA expression in calcified human aortic *** data showed that miR-101-3p levels were increased in the calcified human aortic valves. Using cultured primary HAVICs, we demonstrated that the miR-101-3p mimic promoted calcification and upregulated the osteogenesis pathway, while anti-miR-101-3p inhibited osteogenic differentiation and prevented calcification in HAVICs treated with the osteogenic conditioned medium. Mechanistically, miR-101-3p directly targeted cadherin-11 (CDH11) and Sry-related high-mobility-group box 9 (SOX9), key factors in the regulation of chondrogenesis and osteogenesis. Both CDH11 and SOX9 expressions were downregulated in the calcified human HAVICs. Inhibition of miR-101-3p restored expression of CDH11, SOX9 and ASPN and prevented osteogenesis in HAVICs under the calcific ***-101-3p plays an important role in HAVIC calcification through regulation of CDH11/SOX9 expression. The finding is important as it reveals that miR-1013p may be a potential therapeutic target for calcific aortic valve disease.

关键词： alveolar bone   fiber disorder   Flexcell FX-5000 compression system   occlusal hypofunction   overeruption   periodontal ligament   PLAP-1   asporin  

摘要： Background and ObjectiveTeeth overeruption is a problem of clinical significance, but the underlying mechanism how changes in external occlusal force convert to the periodontium remodeling signals has been a largely under explored domain. And recently, periodontal ligament-associated protein-1 (PLAP-1)/asporin was found to play a pivotal role in maintaining periodontal homeostasis. The aim of this study was to explore the function of PLAP-1 in the periodontally hypofunctional tissue turnover. MethodsAfter extracting left maxillary molars in mice, the left and right mandibular molars were distributed into hypofunction group (HG) and control group (CG), respectively. Mice were sacrificed for radiographic, histological, and molecular biological analyses after 1, 4 and 12 weeks. In vitro, dynamic compression was applied using Flexcell FX-5000 Compression System to simulate intermittent occlusal force. The expression of PLAP1 in loaded and unloaded human periodontal ligament cells (hPDLCs) was compared, and its molecular biological effects were further explored by small interfering RNA (siRNA) targeting PLAP1. ResultsIn vivo, fiber disorder in periodontal ligament (PDL), bone apposition at furcation regions, and bone resorption in alveolar bone were illustrated in the HG compared with the CG. In addition, PLAP-1 positive area decreased significantly in PDL following occlusal unloading. In vitro, the loss of compressive loading relatively downregulated PLAP1 expression, which was essential to promote collagen I but inhibit osterix and osteocalcin expression in hPDLCs. ConclusionsPLAP-1 presumably plays a pivotal role in occlusal force-regulated periodontal homeostasis by facilitating collagen fiber synthesis in hPDLCs and suppressing excessive osteoblast differentiation, further preventing teeth from overeruption. Further evidence in PLAP-1 conditional knockout mice is needed.

关键词： 神经内分泌肿瘤   甲状腺髓样癌   恶性增殖   细胞侵袭   遗传特性   甲状腺癌   MTC   放化疗  

摘要： 甲状腺髓样癌(medullary thyroid cancer, MTC)是一种恶性的神经内分泌肿瘤,由甲状腺C细胞恶性增殖产生,约占所有甲状腺癌的2%~3%。根据疾病的遗传特性,MTC可分为遗传性和散发性两大类。尽管局限性不伴有远处转移的散发性MTC具有良好的预后,10年生存率能够达到70%~80%,但是MTC细胞侵袭性高,很多患者初诊时已是晚期,传统的“手术+放化疗模式”获益有限,10年生存率降到仅有10%。

关键词： Asporin   ECM   Cardiac remodeling   Fibrosis   Ischemia  

摘要： Heart failure is accompanied by adverse cardiac remodeling involving extracellular matrix (ECM). Cardiac ECM acts as a major reservoir for many proteins including growth factors, cytokines, collagens, and proteoglycans. Activated fibroblasts during cardiac injury can alter the composition and activity of these ECM proteins. Through unbiased analysis of a microarray dataset of human heart tissue comparing normal hearts (n = 135) to hearts with ischemic cardiomyopathy (n = 94), we identified Asporin (ASPN) as the top differentially regulated gene (DEG) in ischemic cardiomyopathy;its gene-ontology terms relate closely to fibrosis and cell death. ASPN is a Class I small leucine repeat protein member implicated in cancer, osteoarthritis, and periodontal ligament mineralization. However, its role in cardiac remodeling is still unknown. Here, we initially confirmed our big dataset analysis through cells, mice, and clinical atrial biopsy samples to demonstrate increased Aspn expression after pressure overload or cardiac ischemia/reperfusion injury. We tested the hypothesis that Aspn, being a TGFb1 inhibitor, can attenuate fibrosis in mouse models of cardiac injury. We found that Aspn is released by cardiac fibroblasts and attenuates TGFb signaling. Moreover, Aspn(-/-) mice displayed increased fibrosis and decreased cardiac function after pressure overload by transverse aortic constriction (TAC) in mice. In addition, Aspn protected cardiomyocytes from hypoxia/reoxygenation-induced cell death and regulated mitochondrial bioenergetics in cardiomyocytes. Increased infarct size after ischemia/reperfusion injury in Aspn(-/-) mice confirmed Aspn's contribution to cardiomyocyte viability. Echocardiography revealed greater reduction in left ventricular systolic function post-I/R in the Aspn(-/-) animals compared to wild type. Furthermore, we developed an ASPN-mimic peptide using molecular modeling and docking which when administered to mice prevented TAC-induced fibrosis and preserv

关键词： 椎间盘退变   纤维环修复   生物疗法   组织工程   基因治疗  

摘要： 纤维环损伤与椎间盘退行性疾病(IVDD)的发病密切相关,也是导致椎间盘术后疗效不佳的重要因素,恢复纤维环结构和功能的完整性对缓解甚至逆转IVDD有重要意义。传统的椎间盘纤维环修复策略仅完成物理修复而不能恢复纤维环的生物力学特性,并不是理想的修复方式。近年来,应用生物学方法来修复纤维环取得了一些进展。本文综述了目前对椎间盘纤维环损伤的认识,并介绍了包括细胞移植、纤维环组织工程以及基因治疗在内的各种生物疗法在椎间盘纤维环再生策略方面的最新进展,以期为未来的研究方向选择提供帮助。