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![MMP domain structure and protein fold. (A) The domain organization of each human MMP is illustrated schematically; S, signal peptide; Pro, propeptide; CAT, catalytic domain; F, fibronectin type II repeats; PEX, hemopexin domain; TM, transmembrane domain; GPI, glycophosphatidylinositol membrane anchor; C, cytoplasmic domain; CA, cysteine array; Ig, immunoglobulin-like domain. The flexible, variable length linker between CAT and PEX is shown as a black ribbon. (B) The representative 3D protein fold of proMMP-2 is illustrated; individual domains are colored as in panel A. The flexible linker between CAT and PEX domains, shown as a black dashed line, varies in length among MMPs. The prodomain (gray) inhibits activity by coordinating the catalytic zinc (yellow sphere) and blocking access to substrates. Activation requires proteolysis within the loop indicated by the black arrow, leading to dissociation of the prodomain. Figure was generated with PyMOL (Schrodinger, LLC) from coordinates of PDB ID: 1GXD [Morgunova et al., 2002].](profile/Evette-Radisky/publication/317386396/figure/fig5/AS:660780311666689@1534553711097/MMP-domain-structure-and-protein-fold-A-The-domain-organization-of-each-human-MMP-is.png)
MMP domain structure and protein fold. (A) The domain organization of each human MMP is illustrated schematically; S, signal peptide; Pro, propeptide; CAT, catalytic domain; F, fibronectin type II repeats; PEX, hemopexin domain; TM, transmembrane domain; GPI, glycophosphatidylinositol membrane anchor; C, cytoplasmic domain; CA, cysteine array; Ig, immunoglobulin-like domain. The flexible, variable length linker between CAT and PEX is shown as a black ribbon. (B) The representative 3D protein fold of proMMP-2 is illustrated; individual domains are colored as in panel A. The flexible linker between CAT and PEX domains, shown as a black dashed line, varies in length among MMPs. The prodomain (gray) inhibits activity by coordinating the catalytic zinc (yellow sphere) and blocking access to substrates. Activation requires proteolysis within the loop indicated by the black arrow, leading to dissociation of the prodomain. Figure was generated with PyMOL (Schrodinger, LLC) from coordinates of PDB ID: 1GXD [Morgunova et al., 2002].
Source publication
Matrix metalloproteinases (MMPs) are a family of zinc endopeptidases that cleave nearly all components of the extracellular matrix as well as many other soluble and cell-associated proteins. MMPs have been implicated in normal physiological processes, including development, and in the acquisition and progression of the malignant phenotype. Disappoi...
Context in source publication
Context 1
... MMP catalytic domain is highly conserved among members of the family, and contains key features of the larger metzincin metallopeptidase clan, including the conserved HExxHxxGxxH motif which functions to coordinate the catalytic zinc ion. The MMP catalytic mechanism involves activation of a water molecule by the zinc ion and a conserved Glu residue for nucleophilic attack on the target peptide bond [Tallant et al., 2010;Cerda-Costa and Gomis-Ruth, 2014]. Prior to enzyme activation, the prodomain of the MMP blocks access to the active site cleft (Fig. 1B), usually (with the exception of MMP-26) through interaction of the "cysteine switch" motif containing a conserved PRCGxPD with the catalytic zinc. Activation of the MMP involves disruption of the cysteine switch interaction and cleavage of the prodomain by an activating protease [Rosenblum et al., 2007a;Tallant et al., 2010]. MMP-26, which uniquely possesses a mutated and nonfunctional cysteine switch motif, is also activated proteolytically, although the mechanism by which its zymogen retains latency is not clearly understood [Marchenko et al., 2002]. Many of the MMPs, and all of the transmembrane MMPs, can be activated by furin in the secretory pathway of the cell, while others can be activated once localized to the extracellular space by serine proteases or other MMPs [Ra and Parks, 2007;Tallant et al., ...
Citations
... In this study, we chose MMPs as targets of the effects of the studied polyphenols since these enzymes play a key role in cancer cell invasion by digesting the ECM, supporting cancer cell growth and tumor metastasis [20,21]. In this contest, numerous attempts have been made to generate drugs that are capable of inhibiting MMPs, mainly with the aim of blocking the invasion and metastasis of tumor cells, but the results of clinical trials have proved disappointing, with unwanted side effects and a lack of effectiveness [22]. ...
Polyphenols, the main antioxidants of diet, have shown anti-inflammatory, antioxidant and anticarcinogenic activities. Here, we compared the effects of four polyphenolic compounds on ROS production and on the levels of matrix metalloproteinase (MMP)-2 and -9, which represent important pathogenetic factors of breast cancer. THP-1 differentiated macrophages were activated by LPS and simultaneously treated with different doses of a green tea extract (GTE), resveratrol (RSV), curcumin (CRC) and an olive fruit extract (oliplus). By using the 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, we found that all of the tested compounds showed antioxidant activity in vitro. In addition, GTE, RSV and CRC were able to counteract ROS production induced by H2O2 in THP-1 cells. As assessed by a zymographic analysis of THP-1 supernatants and by an “in-gel zymography” of a pool of sera from patients with breast cancer, the antioxidant compounds used in this study inhibited both the activity and expression of MMP-2 and MMP-9 through different mechanisms related to their structures and to their ability to scavenge ROS. The results of this study suggest that the used antioxidants could be promising agents for the prevention and complementary treatment of breast cancer and other diseases in which MMPs play a pivotal role.
... Metalloproteinases (MP) are responsible in progression of several diseases due to their critical role in remodeling the extracellular matrix (ECM) and involvement in other cellular signaling pathways [1,2]. MPs consist of matrix metalloproteinases (MMP), a disintegrin, and metalloproteinase (ADAM), and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS), and are tightly regulated by activation of their inactive zymogen form and inhibition via tissue inhibitors of metalloproteinases (TIMPs) [1, [3][4][5]. Significant efforts were focused on the discovery of MMP inhibitors as potential therapeutics. However, the initial classes of MMP inhibitors based on small molecules failed in the later stages of clinical trials due to a lack of selectivity and toxicity [1,3]. ...
... Significant efforts were focused on the discovery of MMP inhibitors as potential therapeutics. However, the initial classes of MMP inhibitors based on small molecules failed in the later stages of clinical trials due to a lack of selectivity and toxicity [1,3]. Given the multifaceted role of MPs in ECM and their wide range of substrate specificity [6,7], recent attention was shifted to MP inhibitors by engineering protein-and peptide-based alternatives [8]. ...
Targeting metalloproteinases (MPs) has been the center of attention for developing therapeutics due to their contribution to a wide range of diseases, including cancer, cardiovascular, neurodegenerative disease, and preterm labor. Protein-based MP inhibitors offer higher stability and selectivity, which is critical for developing efficient therapeutics with low off-target effects. Tissue inhibitors of metalloproteinases (TIMPs), natural inhibitors of MPs, and antibodies provide excellent protein scaffolds for engineering selective or multispecific MP inhibitors. Advances in protein engineering and design techniques, such as rational design and directed evolution using yeast display to develop potent MP inhibitors, are discussed, including but not limited to loop grafting, swapping, and counterselective selection.
... responsible for terminating tissue breakdown and bleeding 40 by inhibiting MMP catalytic activity. However, TIMPs also interact with other signaling molecules in cells to regulate differentiation and proliferation, act as chemokines or growth factors, and play an essential role in maintaining tissue homeostasis.[41][42][43] A previous study highlighted that the persistence of TIMP-1 activity at constant levels, coupled with increasing MMP-2 activity, might determine the duration of AUB.44 ...
Objective
To evaluate gene expression associated with vaginal bleeding in the 52‐mg hormonal intrauterine device (IUD) users.
Materials and Methods
We conducted a prospective study involving 100 women seeking to use the 52‐mg hormonal IUD for contraception. We excluded women with a history or current condition of abnormal uterine bleeding and who were unable to attend a 1‐year follow up. Women who expelled the device, removed it for reasons unrelated to vaginal bleeding, or were lost to follow up were discontinued. We collected endometrial biopsies immediately before IUD placement and assessed 20 selected genes using reverse transcription quantitative polymerase chain reaction. Users maintained a uterine bleeding diary for 12 months following IUD insertion. For statistical analysis, participants were categorized into groups with or without vaginal bleeding at 3 and 12 months.
Results
Women with elevated CXCL9 expression had an 8.15‐fold higher likelihood of experiencing vaginal bleeding at 3 months (odds ratio [OR] 8.15, 95% confidence interval [CI] 2.24–29.61, P = 0.001). At 12 months of follow up, women with increased TIMP1 expression had a 2.74‐fold higher chance of experiencing vaginal bleeding (OR 2.74, 95% CI 1.08–6.95, P = 0.033). CXCL9 ≥ 1.5 and IL17A ≥ 0.68 were associated with a higher probability of vaginal bleeding at 3 months, while TIMP1 levels ≥0.943 were linked to an increased risk of bleeding at 12 months.
Conclusion
Users of the 52‐mg hormonal IUD with elevated relative CXCL9 expression face an increased risk of vaginal bleeding at 3‐month follow up, whereas those with heightened TIMP1 expression are more likely to experience vaginal bleeding at 12 months.
... The sequences of the top minimal TIMP variants isolated from screening toward MMP-3cd and MMP-9cd all presented the "CXC" motif, with most of them having Ser at the second position (CSC), at the N-terminus. The CXC motif is known to be critical for binding and inhibition of MMPs as Cys1 of TIMPs contact catalytic His residues of MMPs (Radisky et al., 2017;Raeeszadeh-Sarmazdeh et al., 2020). The three cysteines at positions 1, 3, and 14 are present in all screened minimal TIMP variants from MMP-3cd sorts, which suggests formation of disulfide bonds that are known to improve stability, binding, and inhibition in peptide binders (Moore & Cochran, 2012;Pluda et al., 2021; Figures 2 and 3, Table 1). ...
... The inhibition for minimal TIMP library was performed using the method for tight binding inhibition, as described previously (Radisky et al., 2017;Raeeszadeh-Sarmazdeh & Boder, 2022;Raeeszadeh-Sarmazdeh, Patel, et al., 2019). Briefly, MMP inhibition activities were determined via measuring the rate of MMP cleavage of fluorogenic MMP substrate, Mca-Pro-Leu-Gly-Leu-Dap (Dnp)-Ala-Arg-NH2 (AnaSpec) with a range of the inhibitor variants' concentrations. ...
Overexpression of specific matrix metalloproteinases (MMPs) has a key role in development of several diseases, such as cancer, neurological disorders, and cardiovascular diseases due to their critical role in degradation and remodeling of the extracellular matrix (ECM). Tissue inhibitors of metalloproteinases (TIMPs), a family of four in humans, are endogenous inhibitors of MMPs. TIMPs have a high level of sequence and structure homology, with a broad range of binding and inhibition to the family of MMPs. It is important to identify the key motifs of TIMPs responsible for inhibition of MMPs to develop efficient therapeutics targeting specific MMPs. We used DNA shuffling between the human TIMP family to generate a minimal TIMP hybrid library in yeast to identify the dominant minimal MMP inhibitory regions. The minimal TIMP variants screened toward MMP‐3 and MMP‐9 using fluorescent‐activated cell sorting (FACS). Interestingly, several minimal TIMP variants selected after screening toward MMP‐3cd or MMP‐9cd, with lengths as short as 20 amino acids, maintained or improved binding to MMP‐3 and MMP‐9. The TIMP‐MMP binding dissociation constant (KD), in the nM range, and MMP inhibition constants (Ki), in the pM range, of these minimal TIMP variants were similar to the N‐terminal domain of TIMP‐1 on the yeast surface and in solution indicating the potency of these minimal variants as MMP inhibitors. We further used molecular modeling simulation, and molecular docking of the minimal TIMP variants in complex with MMP‐3cd to understand the binding and inhibition mechanism of these variants.
... Furthermore, enzymes such as matrix metalloproteinases are upregulated in TME. These enzymes not only promote cancer cell invasion and metastasis by clearing the pathways of invading cancer cells but also provide nutrients for vascular tumours (Overall and Kleifeld, 2006;Radisky et al., 2017). In recent decades, substantial advances in the development of TME-responsive therapy have offered promising diagnostic and therapeutic strategies. ...
The treatment of breast cancer (BC) is a serious challenge due to its heterogeneous nature, multidrug resistance (MDR), and limited therapeutic options. Nanoparticle-based drug delivery systems (NDDSs) represent a promising tool for overcoming toxicity and chemotherapy drug resistance in BC treatment. No bibliometric studies have yet been published on the research landscape of NDDS-based treatment of BC. In this review, we extracted data from 1,752 articles on NDDS-based treatment of BC published between 2012 and 2022 from the Web of Science Core Collection (WOSCC) database. VOSviewer, CiteSpace, and some online platforms were used for bibliometric analysis and visualization. Publication trends were initially observed: in terms of geographical distribution, China and the United States had the most papers on this subject. The highest contributing institution was Sichuan University. In terms of authorship and co-cited authorship, the most prolific author was Yu Zhang. Furthermore, Qiang Zhang and co-workers have made tremendous achievements in the field of NDDS-based BC treatment. The article titled “Nanomedicine in cancer therapy: challenges, opportunities, and clinical applications” had the most citations. The Journal of Controlled Release was one of the most active publishers in the field. “Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries” was the most cited reference. We also analysed “hot” and cutting-edge research for NDDSs in BC treatment. There were nine topic clusters: “tumour microenvironment,” “nanoparticles (drug delivery),” “breast cancer/triple-negative breast cancer,” “combination therapy,” “drug release (pathway),” “multidrug resistance,” “recent advance,” “targeted drug delivery”, and “cancer nanomedicine.” We also reviewed the core themes of research. In summary, this article reviewed the application of NDDSs in the treatment of BC.
... 46,47 Tissue inhibitors of matrix metalloproteinase (TIMPs) are responsible for inhibiting the activity of MMPs, and major functional components include TIMP-1 and TIMP-2. 48,49 The expression of MMPs and TIMPs at the mRNA and protein levels was assessed by RT-qPCR and WB, respectively. Notably, the expression levels of MMP-2 and MMP-12 genes were decreased, while the MMP-8, MMP-9 and MMP-25 gene levels were increased after DENV-2 infection, although TIMPs did not change significantly (Fig. 5a). ...
Background:
Dengue virus (DENV) infection during pregnancy increases the risk of adverse fetal outcomes, which has become a new clinical challenge. However, the underlying mechanism remains unknown.
Methods:
The effect of DENV-2 infection on fetuses was investigated using pregnant interferon α/β receptor-deficient (Ifnar1-/-) mice. The histopathological changes in the placentas were analyzed by morphological techniques. A mouse inflammation array was used to detect the cytokine and chemokine profiles in the serum and placenta. The infiltration characteristics of inflammatory cells in the placentas were evaluated by single-cell RNA sequencing.
Findings:
Fetal growth restriction observed in DENV-2 infection was mainly caused by the destruction of the placental vasculature rather than direct damage from the virus in our mouse model. After infection, neutrophil infiltration into the placenta disrupts the expression profile of matrix metalloproteinases, which leads to placental dysvascularization and insufficiency. Notably, similar histopathological changes were observed in the placentas from DENV-infected puerperae.
Interpretation:
Neutrophils play key roles in placental histopathological damage during DENV infection, which indicates that interfering with aberrant neutrophil infiltration into the placenta may be an important therapeutic target for adverse pregnancy outcomes in DENV infection.
Funding:
The National Key Research and Development Plans of China (2021YFC2300200-02 to J.A., 2019YFC0121905 to Q.Z.C.), the National Natural Science Foundation of China (NSFC) (U1902210 and 81972979 to J. A., 81902048 to Z. Y. S., and 82172266 to P.G.W.), and the Support Project of High-level Teachers in Beijing Municipal Universities in the Period of 13th Five-year Plan, China (IDHT20190510 to J. A.).
... birgə (2013) öz tədqiqatlarında MMP9/MPTİ1 nisbətinin xəstəlyin proqnozu ilə əlaqəsi olmadığını göstərmişlər [27]. Abdollahi A. və həmkarları öz tədqiqatlarında MMP2 və MPTİ1 genlərinin ekspressiyası ilə limfa düyünlərinə metastaz arasında asılılıq və gen lərin ekspressiyasında isə etnik və coğrafi fərqlərin olduğunu müəyyən etmişlər [26] Bir neçə tədqiqatda aşkar edilmişdir ki, pro MMP9/MPTİ1 kompleksi MMP9 profermentini həm in vitro, həm də in vivo şəaritdə digər MMP ilə fəallaşmasından qoruyan mexanizmdir [28,29]. ...
Tissue metalloproteinase inhibitors (MPTIs) play an important role in the pathogenesis of breast cancer (BC). Although recent studies have shown the dual role of MPTIs in the development of this pathology, it has been proven that they inhibit the activity of metalloproteinases prevent invasiveness and tumor recurrence. Conducting extensive research towards studying the antitumor effect of these protease inhibitors, especially MPTI-1, may allow the development of new drugs for the treatment of breast cancer.
... MMP inhibitors as cancer therapeutics have been investigated in many studies including one phase III trial, which tested the broad-spectrum MMP inhibitor marimastat in mBC, but no therapeutic benefit was found when it was used after first-line chemotherapy [229]. Although results from clinical trials of small-molecule, broad-spectrum MMP inhibitors as cancer therapeutics were disappointing [230,231], MMP2selective inhibitors were shown to prevent bone metastasis of breast cancer [232]. A recent mega-analysis also suggested that many MMP family members were differentially expressed in patients, and MMP1 and MMP9 were particularly indicated to be potential therapeutic targets [233]. ...
Simple Summary
Metastasis is the root cause of cancer death, responsible for roughly 90% of all cancer-related fatalities. Therefore, treating metastatic cancer is essential for optimal clinical management of these patients. Although there is currently no effective treatment for metastatic breast cancer, high-throughput approaches have identified novel molecules that are critical to tumor growth and metastasis in recent years. Novel therapies that target these molecules, such as immunotherapy, have been evaluated in both preclinical and clinical settings and have proven to be very promising in prolonging survival and relieving symptoms of metastatic disease, thereby enhancing the quality of life of patients. Due to the high intertumoral and intratumoral heterogeneity of cancer, the development of subtype-specific therapeutics and the use of combination treatments to simultaneously target multiple oncogenic signaling pathways may overcome drug resistance and achieve better clinical outcome for metastatic breast cancer.
Abstract
Breast cancer is now the most common cancer worldwide, and it is also the main cause of cancer-related death in women. Survival rates for female breast cancer have significantly improved due to early diagnosis and better treatment. Nevertheless, for patients with advanced or metastatic breast cancer, the survival rate is still low, reflecting a need for the development of new therapies. Mechanistic insights into metastatic breast cancer have provided excellent opportunities for developing novel therapeutic strategies. Although high-throughput approaches have identified several therapeutic targets in metastatic disease, some subtypes such as triple-negative breast cancer do not yet have an apparent tumor-specific receptor or pathway to target. Therefore, exploring new druggable targets in metastatic disease is a high clinical priority. In this review, we summarize the emerging intrinsic therapeutic targets for metastatic breast cancer, including cyclin D-dependent kinases CDK4 and CDK6, the PI3K/AKT/mTOR pathway, the insulin/IGF1R pathway, the EGFR/HER family, the JAK/STAT pathway, poly(ADP-ribose) polymerases (PARP), TROP-2, Src kinases, histone modification enzymes, activated growth factor receptors, androgen receptors, breast cancer stem cells, matrix metalloproteinases, and immune checkpoint proteins. We also review the latest development in breast cancer immunotherapy. Drugs that target these molecules/pathways are either already FDA-approved or currently being tested in clinical trials.
... 32,218 In pursuit of selectivity, RNA-interference (RNAi)-based therapeutics, inhibitory antibodies and peptides, modified versions of endogenous protease inhibitors, and complex natural molecules may serve as potential alternatives for traditional small molecule enzyme inhibitors. 212,[219][220][221][222][223] Proteases can also serve as targets for therapeutic vaccines. In particular, KLK2 and KLK3, which show high expression primarily in the prostate, are attractive targets in this respect. ...
We aimed to study mRNA levels and prognostic impact of all 15 human kallikrein‐related peptidases (KLKs) and their targets, proteinase‐activated receptors (PARs), in surgically treated prostate cancer (PCa). Seventy‐nine patients with localized grade group 2‐4 PCas represented aggressive cases, based on metastatic progression during median follow‐up of 11 years. Eighty‐six patients with similar baseline characteristics, but no metastasis during follow‐up, were assigned as controls. Transcript counts were detected with nCounter technology. KLK12 protein expression was investigated with immunohistochemistry. The effects of KLK12 and KLK15 were studied in LNCaP cells using RNA interference. KLK3, ‐2, ‐4, ‐11, ‐15, ‐10 and ‐12 mRNA, in decreasing order, were expressed over limit of detection (LOD). The expression of KLK2, ‐3, ‐4 and ‐15 was decreased and KLK12 increased in aggressive cancers, compared to controls (P < .05). Low KLK2, ‐3 and ‐15 expression was associated with short metastasis‐free survival (P < .05) in Kaplan‐Meier analysis. PAR1 and ‐2 were expressed over LOD, and PAR1 expression was higher, and PAR2 lower, in aggressive cases than controls. Together, KLKs and PARs improved classification of metastatic and lethal disease over grade, pathological stage and prostate‐specific antigen combined, in random forest analyses. Strong KLK12 immunohistochemical staining was associated with short metastasis‐free and PCa‐specific survival in Kaplan‐Meier analysis (P < .05). Knock‐down of KLK15 reduced colony formation of LNCaP cells grown on Matrigel basement membrane preparation. These results support the involvement of several KLKs in PCa progression, highlighting, that they may serve as prognostic PCa biomarkers.
... It is important to note that inhibitors of MMP activities have given very disappointing results in many clinical trials [453][454][455]. Part of the reason why MMP inhibitors did not succeed as expected is the presence of many members of the MMP family and their overlapping activities [456,457]. In addition to targeting ECM components, many signaling cascades that are activated or downregulated by ECM proteins can be regulated. ...
The extracellular matrix (ECM) is a ubiquitous member of the body and is key to the maintenance of tissue and organ integrity. Initially thought to be a bystander in many cellular processes, the extracellular matrix has been shown to have diverse components that regulate and activate many cellular processes and ultimately influence cell phenotype. Importantly, the ECM's composition, architecture, and stiffness/elasticity influence cellular phenotypes. Under normal conditions and during development, the synthesized ECM constantly undergoes degradation and remodeling processes via the action of matrix proteases that maintain tissue homeostasis. In many pathological conditions including fibrosis and cancer, ECM synthesis, remodeling, and degradation is dysregulated, causing its integrity to be altered. Both physical and chemical cues from the ECM are sensed via receptors including integrins and play key roles in driving cellular proliferation and differentiation and in the progression of various diseases such as cancers. Advances in 'omics' technologies have seen an increase in studies focusing on bidirectional cell-matrix interactions, and here, we highlight the emerging knowledge on the role played by the ECM during normal development and in pathological conditions. This review summarizes current ECM-targeted therapies that can modify ECM tumors to overcome drug resistance and better cancer treatment.
