Investigation of the functions of Matrix Metalloproteinase-8 (MMP-8) in Mammary Carcinoma Cells

Abstract Breast cancer is the most common cancer in the UK today, with incidence rates rising steadily. Prognosis is improving but patients with metastatic disease only have a 15% chance of surviving 5 years beyond prognosis. Therefore, factors influencing the metastatic spread of breast cancer require better understanding. The matrix metalloproteinases (MMPs) are a family of proteases thought to promote metastasis due to their matrix degradation capabilities. MMP-8 however has been discovered to be anti-tumourigenic in many cancers, and anti-metastatic in breast cancer. This occurs through unknown mechanisms, so this work sought to gain further insight into the functional effects of MMP-8. In the literature it has been suggested that MMP-8 is tumour protective through its role in regulating innate immune responses and preventing chronic inflammation. This occurs through activational cleavage of a pro-inflammatory chemokine, Interleukin-8 (IL-8). Using an in vitro over-expression model it is shown in this thesis that MMP-8 reduced 2D random migration and scratch wound closure. It also increased cell adhesion and reduced colony formation and prevented primary tumour growth in mice. These data indicate an anti-tumourigenic role for MMP-8. Biochemically, MMP-8 induced expression of IL-8 in mammary carcinoma cells, and also the expression of a pro-inflammatory cytokine IL-6, dependent on its catalytic activity. This required NFĸB signalling and IL-6 also required Protease Activated receptor-2. This occurred following transient expression of MMP-8, and also in rare stably transfected clones that expressed MMP-8 long-term. However, wild-type MMP-8 was not tolerated by breast cancer cells and was epigenetically silenced, potentially as a mechanism to overcome growth inhibitory effects exerted by wild-type MMP-8. In these rare “long-term” MMP-8 expressing cells phenotypic alterations occurred, including elevated IL-6 and IL- 8 expression independent of MMP-8, and a self-reinforcing loop between MMP- 8, IL-6 and IL-8. 3 This pathway may contribute to the anti-tumourigenic and metastasis suppressive effects of MMP-8, or it may represent a cellular response to overcome the anti-tumour actions of the protease

Pleiotropic functions of the tumor- and metastasis-suppressing Matrix Metalloproteinase-8 in mammary cancer in MMTV-PyMT transgenic mice

Matrix metalloproteinase-8 (MMP-8; neutrophil collagenase) is an important regulator of innate immunity which has onco-suppressive actions in numerous tumor types

To which world regions does the valence–dominance model of social perception apply?

Over the past 10 years, Oosterhof and Todorov’s valence–dominance model has emerged as the most prominent account of how people evaluate faces on social dimensions. In this model, two dimensions (valence and dominance) underpin social judgements of faces. Because this model has primarily been developed and tested in Western regions, it is unclear whether these findings apply to other regions. We addressed this question by replicating Oosterhof and Todorov’s methodology across 11 world regions, 41 countries and 11,570 participants. When we used Oosterhof and Todorov’s original analysis strategy, the valence–dominance model generalized across regions. When we used an alternative methodology to allow for correlated dimensions, we observed much less generalization. Collectively, these results suggest that, while the valence–dominance model generalizes very well across regions when dimensions are forced to be orthogonal, regional differences are revealed when we use different extraction methods and correlate and rotate the dimension reduction solution.C.L. was supported by the Vienna Science and Technology Fund (WWTF VRG13-007); L.M.D. was supported by ERC 647910 (KINSHIP); D.I.B. and N.I. received funding from CONICET, Argentina; L.K., F.K. and Á. Putz were supported by the European Social Fund (EFOP-3.6.1.-16-2016-00004; ‘Comprehensive Development for Implementing Smart Specialization Strategies at the University of Pécs’). K.U. and E. Vergauwe were supported by a grant from the Swiss National Science Foundation (PZ00P1_154911 to E. Vergauwe). T.G. is supported by the Social Sciences and Humanities Research Council of Canada (SSHRC). M.A.V. was supported by grants 2016-T1/SOC-1395 (Comunidad de Madrid) and PSI2017-85159-P (AEI/FEDER UE). K.B. was supported by a grant from the National Science Centre, Poland (number 2015/19/D/HS6/00641). J. Bonick and J.W.L. were supported by the Joep Lange Institute. G.B. was supported by the Slovak Research and Development Agency (APVV-17-0418). H.I.J. and E.S. were supported by a French National Research Agency ‘Investissements d’Avenir’ programme grant (ANR-15-IDEX-02). T.D.G. was supported by an Australian Government Research Training Program Scholarship. The Raipur Group is thankful to: (1) the University Grants Commission, New Delhi, India for the research grants received through its SAP-DRS (Phase-III) scheme sanctioned to the School of Studies in Life Science; and (2) the Center for Translational Chronobiology at the School of Studies in Life Science, PRSU, Raipur, India for providing logistical support. K. Ask was supported by a small grant from the Department of Psychology, University of Gothenburg. Y.Q. was supported by grants from the Beijing Natural Science Foundation (5184035) and CAS Key Laboratory of Behavioral Science, Institute of Psychology. N.A.C. was supported by the National Science Foundation Graduate Research Fellowship (R010138018). We acknowledge the following research assistants: J. Muriithi and J. Ngugi (United States International University Africa); E. Adamo, D. Cafaro, V. Ciambrone, F. Dolce and E. Tolomeo (Magna Græcia University of Catanzaro); E. De Stefano (University of Padova); S. A. Escobar Abadia (University of Lincoln); L. E. Grimstad (Norwegian School of Economics (NHH)); L. C. Zamora (Franklin and Marshall College); R. E. Liang and R. C. Lo (Universiti Tunku Abdul Rahman); A. Short and L. Allen (Massey University, New Zealand), A. Ateş, E. Güneş and S. Can Özdemir (Boğaziçi University); I. Pedersen and T. Roos (Åbo Akademi University); N. Paetz (Escuela de Comunicación Mónica Herrera); J. Green (University of Gothenburg); M. Krainz (University of Vienna, Austria); and B. Todorova (University of Vienna, Austria). The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.https://www.nature.com/nathumbehav/am2023BiochemistryGeneticsMicrobiology and Plant Patholog

To which world regions does the valence–dominance model of social perception apply?

Over the past 10 years, Oosterhof and Todorov’s valence–dominance model has emerged as the most prominent account of how people evaluate faces on social dimensions. In this model, two dimensions (valence and dominance) underpin social judgements of faces. Because this model has primarily been developed and tested in Western regions, it is unclear whether these findings apply to other regions. We addressed this question by replicating Oosterhof and Todorov’s methodology across 11 world regions, 41 countries and 11,570 participants. When we used Oosterhof and Todorov’s original analysis strategy, the valence–dominance model generalized across regions. When we used an alternative methodology to allow for correlated dimensions, we observed much less generalization. Collectively, these results suggest that, while the valence–dominance model generalizes very well across regions when dimensions are forced to be orthogonal, regional differences are revealed when we use different extraction methods and correlate and rotate the dimension reduction solution

A multi-country test of brief reappraisal interventions on emotions during the COVID-19 pandemic.

The COVID-19 pandemic has increased negative emotions and decreased positive emotions globally. Left unchecked, these emotional changes might have a wide array of adverse impacts. To reduce negative emotions and increase positive emotions, we tested the effectiveness of reappraisal, an emotion-regulation strategy that modifies how one thinks about a situation. Participants from 87 countries and regions (n = 21,644) were randomly assigned to one of two brief reappraisal interventions (reconstrual or repurposing) or one of two control conditions (active or passive). Results revealed that both reappraisal interventions (vesus both control conditions) consistently reduced negative emotions and increased positive emotions across different measures. Reconstrual and repurposing interventions had similar effects. Importantly, planned exploratory analyses indicated that reappraisal interventions did not reduce intentions to practice preventive health behaviours. The findings demonstrate the viability of creating scalable, low-cost interventions for use around the world

Loss of MMP-8 accelerates tumor onset; promotes progression, tumor size and lung macrometastases in the MMTV-PyMT model.

<p>A) Detection of first palpable tumors of MMTV-PyMT; Mmp8 wild-type (WT, n = 17), MMTV-PyMT; Mmp8 heterozygote (HET, n = 30) and MMTV-PyMT; Mmp8 null (KO, n = 10) mice. Mean ± sem, 2-tailed unpaired t-test; * p < 0.05; ** p < 0.01. B) Tumor progression determined by palpation of all 10 mammary glands from day 21 to day 98 for MMTV-PyMT; Mmp8 wild-type (WT, n = 26), heterozygote (HET, n = 34) and null (KO, n = 21) mice. C) Tumor size distribution (0, < 0.5, 0.5-1, >1 cm) at 10 weeks of age for MMTV-PyMT; Mmp8 wild-type (WT, n = 26), MMTV-PyMT; Mmp8 heterozygote (HET, n = 34) and MMTV-PyMT; Mmp8 null (KO, n = 21) mice; and at 14 weeks of age for MMTV-PyMT; Mmp8 wild-type (WT, n = 22), MMTV-PyMT; Mmp8 heterozygote (HET, n = 30) and MMTV-PyMT; Mmp8 null (KO, n = 16) mice. Chi-square test, * p < 0.01. D) Number of lung macrometastases assessed at 14 weeks of age for MMTVPyMT; Mmp8 wild-type (WT, n = 4), MMTV-PyMT; Mmp8 heterozygote (HET, n = 4) and MMTV-PyMT; Mmp8 null (KO, n = 2) mice. Left panel of representative pictures depicts the top and bottom of all lung lobes (1 lobe of the left lung, 4 lobes of the right lung). Black solid arrows = smaller macrometastases, white solid arrows = larger macrometastases. Mean ± sem, 2-tailed unpaired t-test; ** p < 0.01.</p

Altered MMTV-PyMT tumor vascularity in Mmp8-null mice.

<p>Vascular density of tumors in early and late stage tumors as determined by Endomucin staining. The number of mice used for analyses were as following: 8 weeks; MMTV-PyMT; Mmp8 wild-type (WT, n = 4), MMTV-PyMT; Mmp8 heterozygote (HET, n = 5), MMTV-PyMT; Mmp8 null (KO, n = 5), and 10 weeks; MMTV-PyMT; Mmp8 wild-type (WT, n = 4), MMTV-PyMT; Mmp8 heterozygote (HET, n = 6) and MMTV-PyMT; Mmp8 null (KO, n = 4). Representative pictures of 10 week tumors are shown.* p < 0.05, ** p < 0.01.</p

MMP-8 ablation perturbs the tumor protease web at the protein level.

<p>A) MMP-3 protein expression in tumor tissue at 14 weeks of age as determined by western blotting. Quantification of 3 biological replicates per genotype, normalized to Hsc70 and compared to PyMT wild-type (WT). 2-tailed unpaired t-test, mean ± sem. B) Tumor expression of MMP-2 and MMP-9 forms by gelatin Zymography at 10 and 14 weeks of age. 2-tailed unpaired t-test; AU, arbitrary unit; FC, fold change; ns, non-significant</p

Additional file 1: Table S1. of Loss of MMP-8 in ductal carcinoma in situ (DCIS)-associated myoepithelial cells contributes to tumour promotion through altered adhesive and proteolytic function

Summary of breast tissue examined for MMP8. (DOC 27 kb

Additional file 5: Figure S3. of Loss of MMP-8 in ductal carcinoma in situ (DCIS)-associated myoepithelial cells contributes to tumour promotion through altered adhesive and proteolytic function

(i) Densitometry quantifying pSMAD2 versus tSMAD2 normalised to the loading control. MECs transfected with MMP-8 WT show a marked reduction of pSMAD2 compared to Empty Vector and MMP-8 EA at 5 minutes. (ii) Densitometry quantifying pSMAD2 versus tSMAD2 normalised to the loading control. MECs transfected with siRNA to MMP-8 demonstrated a markedly stronger pSMAD2 signal compared to control siRNA (siLUC). (TIF 336 kb