scholarly journals The Sp1-mediaded allelic regulation of MMP13 expression by an ESCC susceptibility SNP rs2252070

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Meng Shi ◽  
Jianhong Xia ◽  
Huaixin Xing ◽  
Wenjun Yang ◽  
Xiangyu Xiong ◽  
...  
Author(s):  
Kentaro Takeuchi ◽  
Hiroyasu Ogawa ◽  
Norishige Kuramitsu ◽  
Keisuke Akaike ◽  
Atsushi Goto ◽  
...  

2021 ◽  
Author(s):  
J Krüger ◽  
M Breunig ◽  
T Seufferlein ◽  
M Hohwieler ◽  
A Kleger

Life ◽  
2020 ◽  
Vol 10 (5) ◽  
pp. 58
Author(s):  
Aida Martinez-Sanchez ◽  
Stefano Lazzarano ◽  
Eshita Sharma ◽  
Helen Lockstone ◽  
Christopher L. Murphy

MicroRNAs (miRNAs) play key roles in cartilage development and homeostasis and are dysregulated in osteoarthritis. MiR-145 modulation induces profound changes in the human articular chondrocyte (HAC) phenotype, partially through direct repression of SOX9. Since miRNAs can simultaneously silence multiple targets, we aimed to identify the whole targetome of miR-145 in HACs, critical if miR-145 is to be considered a target for cartilage repair. We performed RIP-seq (RNA-immunoprecipitation and high-throughput sequencing) of miRISC (miRNA-induced silencing complex) in HACs overexpressing miR-145 to identify miR-145 direct targets and used cWords to assess enrichment of miR-145 seed matches in the identified targets. Further validations were performed by RT-qPCR, Western immunoblot, and luciferase assays. MiR-145 affects the expression of over 350 genes and directly targets more than 50 mRNAs through the 3′UTR or, more commonly, the coding region. MiR-145 targets DUSP6, involved in cartilage organization and development, at the translational level. DUSP6 depletion leads to MMP13 upregulation, suggesting a contribution towards the effect of miR-145 on MMP13 expression. In conclusion, miR-145 directly targets several genes involved in the expression of the extracellular matrix and inflammation in primary chondrocytes. Thus, we propose miR-145 as an important regulator of chondrocyte function and a new target for cartilage repair.


Author(s):  
Sean K Bedingfield ◽  
Fang Yu ◽  
Danielle D. Liu ◽  
Meredith A. Jackson ◽  
Lauren E. Himmel ◽  
...  

AbstractOsteoarthritis (OA) is a debilitating and prevalent chronic disease, but there are no approved disease modifying OA drugs (DMOADs), only pharmaceuticals for pain management. OA progression, particularly for post-traumatic osteoarthritis (PTOA), is associated with inflammation and enzymatic degradation of the extracellular matrix. In particular, Matrix Metalloproteinase 13 (MMP13) breaks down collagen type 2 (CII), a key structural component of cartilage extracellular matrix, and consequently, matrix degradation fragments perpetuate inflammation and a degenerative cycle that leads to progressive joint pathology. Here, we tested targeted delivery of endosome-escaping, MMP13 RNA interference (RNAi) nanoparticles (NPs) as a DMOAD. The new targeting approach pursued here deviates from the convention of targeting specific cell types (e.g., through cell surface receptors) and instead leverages a monoclonal antibody (mAbCII) that targets extracellular CII that becomes uniquely accessible at early OA focal defects. Targeted mAbCII-siNPs create an in situ NP depot for retention and potent activity within OA joints. The mAbCII-siNPs loaded with MMP13 siRNA (mAbCII-siNP/siMMP13) potently suppressed MMP13 expression (95% silencing) in TNFα-stimulated chondrocytes in vitro, and the targeted mAbCII-siNPs had higher binding to trypsin-damaged porcine cartilage than untargeted control NPs. In an acute mechanical injury mouse model of PTOA, mAbCII-siNP/siMMP13 achieved 80% reduction in MMP13 expression (p = 0.00231), whereas a non-targeted control achieved only 55% silencing. In a more severe, PTOA model, weekly mAbCII-siNP/siMMP13 long-term treatment provided significant protection of cartilage integrity (0.45+/− .3 vs 1.6+/−.5 on the OARSI scale; p=0.0166), and overall joint structure (1.3+/−.6 vs 2.8+/−.2 on the Degenerative Joint Disease scale; p<0.05). Intra-articular mAbCII-siNPs better protected articular cartilage (OARSI score) relative to either single or weekly treatment with the clinical gold stand steroid treatment methylprednisolone. Finally, multiplexed gene expression analysis of 254 inflammation-related genes showed that MMP13 inhibition suppressed clusters of genes associated with tissue restructuring, angiogenesis (associated with synovial inflammation and thickening), innate immune response, and proteolysis. This work establishes the new concept of targeting unique local extracellular matrix signatures to sustain retention and increase delivery efficacy of biologics with intracellular activity and also validates the promise of MMP13 RNAi as a DMOAD in a clinically-relevant therapeutic context. Abstract Figure:PTOA targeted delivery of MMP13 siRNA to block disease progressionThe top left schematic illustrates the progression (left to right) from healthy knee joint, through inflammation induction following traumatic injury, to cartilage loss and degenerative joint disease (including synovial response). Degradation of cartilage enhances inflammation, inducing a degenerative cycle (middle right). The bottom of the graphic illustrates the concept of the matrix targeted nanocarriers for enhanced retention and activity of MMP13 siRNA at sites of cartilage injury.


2021 ◽  
Author(s):  
Chan XI ◽  
Chuanxi XIONG ◽  
Huiping WANG ◽  
Yuanjun LIU ◽  
Suju Luo

Abstract Matrix metalloproteinase 13 (MMP13) is a zinc-containing endopeptidase secreted by keratinocytes and skin fibroblasts and participates in many inflammatory diseases. Drugs for retinoic acid include tazarotene and acitretin. Tazarotene/acitretin and narrow-band ultraviolet B (NB-UVB) irradiation are used as a general treatment for psoriasis. However, their impact on MMP13 expression has yet to be determined. In this study, we measured the expression of MMP13 in patients with psoriasis, and investigated the effects of tazarotene and/or NB-UVB on MMP13 expression in a mouse model of psoriasis. After exposure to acitretin and/or NB-UVB, immortalized human HaCaT keratinocytes were analyzed for viability and MMP13 expression. Our results showed that MMP13 protein levels increased in skin lesions and serum samples in patients with psoriasis. Treatment with acitretin and NB-UVB irradiation alone or in combination suppressed cell viability and MMP13 expression in HaCaT cells. Consistently, tazarotene treatment and/or NB-UVB irradiation attenuated imiquimod-induced psoriasis-like dermatitis and inhibited MMP13 expression in a mouse model. Taken together, these results indicate that tazarotene/acitretin and NB-UVB irradiation can inhibit the expression of MMP13 in keratinocytes and psoriasis mouse models. Targeting MMP13 may represent a promising therapeutic strategy against psoriasis.


1993 ◽  
Vol 4 (5) ◽  
pp. 1140-1150
Author(s):  
P G Blake ◽  
J Madrenas ◽  
P F Halloran

Ly-6 is a multigene family of murine polymorphic cell membrane proteins that are glycosydlphosphatidylinositol anchored, widely expressed on lymphoid tissue, and homologous to the recently described human CD59. An unexpected feature of Ly-6 is its high level of expression in the kidney. This renal expression and its interferon (IFN)-gamma inducibility in murine strains expressing different Ly-6 haplotypes were studied with monoclonal antibodies and cDNA probes that recognize Ly-6A/E and Ly-6C. Ly-6 expression was much more extensive in the kidney than in other parenchymal organs. Ly-6A.1/E.2 was extensively expressed on vascular endothelium and on tubular epithelium, particularly in the distal nephron. Pattern of expression differed between strains expressing A and E alleles. Ly-6C was not detected by monoclonal antibodies but was detected by oligonucleotide-specific probes. Treatment with recombinant IFN-gamma or IFN-inducing agents increased Ly-6 expression markedly, particularly on the luminal aspect of the proximal tubular epithelium, where Ly-6A/E became prominent. This luminal expression is typical for glycosydlphosphatidylinositol-anchored proteins but contrasts with that of other molecules, such as major histocompatibility classes I and II, which are generally expressed on the basolateral surface of the tubular epithelium. Up-regulation occurred within 6 h of IFN-gamma treatment and returned to normal by 48 h. Similar up-regulation of Ly-6 was seen in murine lupus nephritis and in mercuric chloride nephropathy. The characteristics of renal Ly-6, such as its IFN-gamma responsiveness, endothelial and tubular expression, polymorphism, strong antigenicity, and possible allelic regulation, make it a candidate to be a target molecule in alloresponses. The renal expression of Ly-6 is similar to that of CD59 in the human kidney, supporting the suggestion that these proteins are closely related.


2021 ◽  
Author(s):  
Antonio Lentini ◽  
Huaitao Cheng ◽  
Joyce Carol Noble ◽  
Natali Papanicolaou ◽  
Christos Coucoravas ◽  
...  

X-chromosome inactivation (XCI) and upregulation (XCU) are the major opposing chromosome-wide modes of gene regulation that collectively achieve dosage compensation in mammals, but the regulatory link between the two remains elusive. Here, we use allele-resolved single-cell RNA-seq combined with chromatin accessibility profiling to finely dissect the separate effects of XCI and XCU on RNA levels during mouse development. We uncover that balanced X dosage is flexibly attained through expression tuning by XCU in a sex- and lineage-specific manner along varying degrees of XCI and across developmental and cellular states. Male blastomeres achieve XCU upon zygotic genome activation while females experience two distinct waves of XCU, upon imprinted- and random XCI, and ablation of Xist impedes female XCU. Contrary to widely established models of mammalian dosage compensation, naïve female embryonic cells carrying two active X chromosomes do not exhibit upregulation but express both alleles at basal level, yet collectively exceeding the RNA output of a single hyperactive allele. We show, in vivo and in vitro, that XCU is kinetically driven by X-specific modulation of transcriptional burst frequency, coinciding with increased compartmentalization of the hyperactive allele. Altogether, our data provide unprecedented insights into the dynamics of mammalian XCU, prompting a revised model of the chain in events of allelic regulation by XCU and XCI in unitedly achieving stable cellular levels of X-chromosome transcripts.


2020 ◽  
Author(s):  
Aida Martinez-Sanchez ◽  
Stefano Lazzarano ◽  
Eshita Sharma ◽  
Chris L. Murphy

ABSTRACTObjectiveMicroRNAs play a key role in biological processes, including cartilage development and homeostasis and are dysregulated in many diseases, including osteoarthritis. MiR-145 modulation induces profound changes in the human articular chondrocyte (HAC) phenotype, partially through direct repression of SOX9. Since miRNAs can simultaneously silence multiple targets, we aimed to identify the whole targetome of miR-145 in HACs. This information is critical if miR-145 is to be considered a target for cartilage repair.MethodsRIP-seq (RNA-immunoprecipitation plus HT-sequencing) of miRISC (miRNA-induced silencing complex) was performed in HACs overexpressing miR-145 to identify miR-145 direct targets. The motif discovery method cWords was used to assess enrichment on miR-145 seed matches in the identified targets. RT-qPCR, Western (immuno-)blot and luciferase assays were used to validate miRNA-target interactions.ResultsMiR-145 overexpression affects the expression of over 350 genes and directly targets more than 50 mRNAs through the 3’UTR or, more commonly, the coding region.We also demonstrate that miR-145 targets DUSP6, involved in cartilage organization and development, at the translational level. DUSP6 depletion using specific siRNAs lead to MMP13 up-regulation, suggesting that miR-145-mediated DUSP6 depletion contributes to the effect of miR-145 on MMP13 expression.ConclusionWe demonstrate that miR-145 directly targets several genes in primary chondrocytes including those involved in the expression of the extracellular matrix and inflammation. Thus, we propose miR-145 as an important regulator of chondrocyte function and a new target for cartilage repair.


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