Maria Ines Almeida

University of Porto | UP

Background The vast and promising class of long non-coding RNAs (lncRNAs) has been under investigation for distinct therapeutic applications. Nevertheless, their role as molecular drivers of bone regeneration remains poorly studied. The lncRNA H19 mediates osteogenic differentiation of Mesenchymal Stem/Stromal Cells (MSCs) through the control of in...

Background Proteins, protein-encoding transcripts and microRNAs have been investigated as molecular drivers of bone regeneration but few reached clinical relevance, while the vast and promising class of long non-coding RNAs (lncRNAs) remains poorly studied. The lncRNA H19 mediates osteogenic differentiation of Mesenchymal Stem/Stromal Cells (MSCs)...

Neuroinflammation is increasingly recognized as playing a critical role in depression. Early-life stress exposure and constitutive differences in glucocorticoid responsiveness to stressors are two key risk factors for depression, but their impacts on the inflammatory status of the brain is still uncertain. Moreover, there is a need to identify spec...

Multiple myeloma (MM) is the second most frequent hematological disease and can cause skeletal osteolytic lesions. This study aims to evaluate the expression of circulating microRNAs (miRNAs) in MM patients and to correlate those levels with clinicopathological features, including bone lesions. A panel of miRNAs associated with MM onset and progres...

Melanoma is the deadliest form of skin cancer, primarily due to its high metastatic propensity and therapeutic resistance in advanced stages. The frequent inactivation of the p53 tumour suppressor protein in melanomagenesis may predict promising outcomes for p53 activators in melanoma therapy. Herein, we aimed to investigate the antitumor potential...

The burden of chronic inflammatory diseases of joints and the spine is increasing with population ageing and unhealthy lifestyles. Articular cartilage and intervertebral discs (IVDs) are avascular and aneural tissues with abundant extracellular matrix, low cell density, and reduced regenerative capacity after damage or degeneration. The most advanc...

Growing evidences suggest that sustained neuroinflammation, caused by microglia overactivation, is implicated in the development and aggravation of several neurological and psychiatric disorders. In some pathological conditions, microglia produce increased levels of cytotoxic and inflammatory mediators, such as tumor necrosis factor alpha (TNF-α),...

Background The tight coupling between osteoblasts and osteoclasts is essential to maintain bone homeostasis. Deregulation of this process leads to loss and deterioration of the bone tissue causing diseases, such as osteoporosis. MicroRNAs are able to control cell differentiation of bone cells and thus, have been explored as therapeutic tools. In th...

Molecular biology: decoding the dark matter of osteoporosis Genomic ‘dark matter’ could improve the diagnosis and prognosis of patients with osteoporosis, as well as providing new drug targets for the disease. Only 2.3% of the human genome is translated into protein; the function of the remainder had been unclear, but recent studies suggest some of...

Rheumatoid arthritis (RA) is a systemic disease that affects the osteoarticular system, associated with bone fragility and increased risk of fractures. Herein, we aimed to characterize the systemic impact of the rat collagen-induced arthritis (CIA) model and explore its combination with femoral bone defect (FD). The impact of CIA on endogenous mese...

Half of human cancers harbor TP53 mutations that render p53 inactive as a tumor suppressor. In these cancers, reactivation of mutant p53 (mutp53) through restoration of wild-type-like function constitutes a valuable anticancer therapeutic strategy. In order to search for mutp53 reactivators, a small library of tryptophanol-derived oxazoloisoindolin...

The normal bone regeneration process is a complex and coordinated series of events involving different cell types and molecules. However, this process is impaired in critical-size/large bone defects, with non-unions or delayed unions remaining a major clinical problem. Novel strategies are needed to aid the current therapeutic approaches. Mesenchym...

Depression is a highly prevalent psychiatric condition, with over 300 million sufferers, and is an important comorbidity for other conditions, like cardiovascular disorders or diabetes. Therapy is largely based on psychotherapy and/or pharmacological intervention, particularly aimed at altering neurotransmitter levels in the central nervous system,...

Autism comprises a complex and heterogeneous spectrum of neurodevelopmental disorders, usually termed autism spectrum disorders (ASD). It is more prevalent in males than females, and genetic and environmental factors are believed to account in similar percentages to the development of ASD. In recent years, the contribution of inflammation and infla...

Non-destructive measurements of cell persistence and gene expression are crucial for longitudinal research studies and for prognostic assessment of cell therapies. Here we describe S-MiRAGE, a platform that utilizes small secreted RNA molecules as sensitive and quantitatively accurate reporters of cellular processes. S-MiRAGE allows cellular number...

Extracellular vesicles (EV), in particular exosomes, have been the object of intense research, due to their potential to mediate intercellular communication, modulating the phenotype of target cells. The natural properties and functions of EV are being exploited as biomarkers for disease diagnosis and prognosis, and as nano-bio-carriers for the dev...

Bone injury healing is an orchestrated process that starts with an inflammatory phase followed by repair and remodelling of the bone defect. The initial inflammation is characterized by local changes in immune cell populations and molecular mediators, including microRNAs (miRNAs). However, the systemic response to bone injury remains largely unchar...

Non-destructive measurements of cell persistence and gene expression are crucial for longitudinal research studies and for prognostic assessment of cell therapies. Here we describe S-MiRAGE, a platform that utilizes small secreted RNA molecules as sensitive and quantitatively accurate reporters of cellular processes. We demonstrate the utility of S...

The cancer-risk-associated rs6983267 single nucleotide polymorphism (SNP) and the accompanying long noncoding RNACCAT2in the highly amplified 8q24.21 region have been implicated in cancer predisposition, although causality has not been established. Here, using allele-specificCCAT2transgenic mice, we demonstrate thatCCAT2overexpression leads to spon...

Orchestration of bone repair processes requires crosstalk between different cell populations, including immune cells and mesenchymal stem/stromal cells (MSC). Extracellular vesicles (EV) as mediators of these interactions remain vastly unexplored. Here, we aimed to determine the mechanism of MSC recruitment by Dendritic Cells (DC), hypothesising th...

Macrophages are a main component of atherosclerotic plaques. Recent studies suggest that pro-inflammatory M1 macrophages are pro-atherogenic while M2 macrophages promote plaque stability. Moreover, toll-like receptor signalling pathways are implicated in atherosclerotic plaque formation, evolution and regression. We propose microRNAs as key regulat...

Human primary monocyte culture, differentiation into macrophages, and polarization. A) Brightfield microscopy image of macrophages differentiated from isolated blood monocytes at day 10 of culture (magnification of 200X). Scale bar 50μm. B) Flow cytometry analysis of CD14 marker in macrophages (right panel). Cells labeled with matching isotype were...

Transfection efficiency of miR-195 mimics in THP-1 macrophages. Transfection efficiency was evaluated by RT-qPCR and compared with SCR control (n = 3). Statistical significance: ****p<0.0001. (TIF)

HUASMC characterization by flow cytometry and inverted fluorescence microscopy. A) Characterization of HUASMC by flow cytometry. Histograms represent flow cytometry results for CD10, CD34 (negative markers) and CD90 (positive marker). B) Evaluation of HUASMC markers by inverted fluorescence microscopy. Cells show positive staining for α-SMA (on top...

Intracellular TLR4 levels in THP-1 macrophages. Intracellular TLR4 levels were measured by flow cytometry (n = 3). Statistical significance: *p<0.05. (TIF)

Quantification of protein p54 JNK, p46 JNK and p38 MAPK levels. A) Representative western blot showing protein phosphorylated levels following LPS and IFN-γ stimulation at different time points (0, 5, 15, 30, 60, 120 minutes). B) Total protein expression of JNK isoforms and p38 MAPK was evaluated by western blot and levels were normalized to α-tubu...

Primers used for RT-qPCR. (DOCX)

Sequence of mature human miR-195-5p according to miRBase, primers used to generate PGL3-constructs for luciferase assays and to generate deletions in the predicted miRNA-binding site. Restriction site for endonuclease is underlined. (DOCX)

In recent years, evidence supporting a link between inflammation and neuropsychiatric disorders has been mounting. Autism spectrum disorders (ASD) and schizophrenia share some clinical similarities which we hypothesize might reflect the same biological basis, namely, in terms of inflammation. However, the diagnosis of ASD and schizophrenia relies s...

The regulation of microRNA (miRNA) biogenesis, function and degradation involves a range of mechanisms, including interactions with RNA-binding proteins. The potential contribution of regulatory miRNAs to the expression of these RNA interactor proteins that could control other miRNAs expression is still unclear. Here we demonstrate a regulatory cir...

Orchestration of bone repair processes requires crosstalk between different cell populations, including immune cells and mesenchymal stem/stromal cells (MSC). Extracellular vesicles (EV) as mediators of these interactions remain vastly unexplored. Here, we aimed to determine the mechanism of MSC recruitment by Dendritic Cells (DC), hypothesising th...

Significance: This study assesses the effects of bone marrow-derived mesenchymal stem cells (MSCs) transplanted systemically for intervertebral (IVD) regeneration into a rat IVD lesion model. It demonstrates systemic MSC transplantation is effective in inhibiting disc degeneration and herniation in situ. Moreover, a systemic immunomodulatory effec...

Inflammation is a complex and highly regulated biological process, crucial for a variety of functions in the human body, from host response against infectious agents to initiation of repair/regeneration of injured tissues. In the context of tissue repair, the action of different immune cell populations and their interplay with tissue specific cells...

Extracellular vesicles (EVs) have been a growing interest of the scientific community in recent years due to the wide possibilities of their evaluation as biomarkers of disease, and their potential to be used as therapeutic agents or vehicles. EVs that circulate in plasma carry proteins and nucleic acids, potentially to distant locations in the bod...

Editorial summary miR-143 and miR-145 have been widely described as tumor suppressors. Recent findings suggest these microRNAs (miRNAs) have a critical role that affects the stroma rather than epithelial cells. Understanding the part played by the miR-143/miR-145 cluster in the tumor microenvironment is essential for the development of future miRNA...

Mesenchymal Stromal/Stem Cells (MSC) are currently being explored in diverse clinical applications, including regenerative therapies. Their contribution to regeneration of bone fractures is dependent on their capacity to proliferate, undergo osteogenesis and induce angiogenesis. This study aimed to uncover microRNAs capable of concomitantly regulat...

OBJECTIVE: MicroRNA (miRNA) expression profile can be used as prognostic marker for human cancers. We aim to explore the significance of miRNAs in colorectal cancer (CRC) metastasis. DESIGN: We performed miRNA microarrays using primary CRC tissues from patients with and without metastasis, and validated selected candidates in 85 CRC samples by quan...

Background. Posttranscriptional regulation is a critical control point for the expression of genes that promote or retard tumor growth. The mRNA-binding protein hnRNP A1 is required for the processing of miR-18a, an inhibitor of k-ras expression. In this study we hypothesized that downmodulation of hnRNP A1 by miR-25 and miR-15a controls the expres...

Unlabelled: Development of improved RNA interference-based strategies is of utmost clinical importance. Although siRNA-mediated silencing of EphA2, an ovarian cancer oncogene, results in reduction of tumor growth, we present evidence that additional inhibition of EphA2 by a microRNA (miRNA) further "boosts" its antitumor effects. We identified miR...

The Epstein-Barr virus (EBV) is associated with a large spectrum of lymphoproliferative diseases. Traditional methods of EBV detection include the immunohistochemical identification of viral proteins and DNA probes to the viral genome in tumoral tissue. The present study explored the detection of the EBV genome, using the BALF5 gene, in the bone ma...

Trastuzumab, a humanized monoclonal antibody directed against the extracellular domain of the HER2 oncoprotein, can effectively target HER2-positive breast cancer through several mechanisms. Although the effects of trastuzumab on cancer cell proliferation, angiogenesis and apoptosis have been investigated in depth, the effect of trastuzumab on micr...

Effects of trastuzumab on miR-194 expression in the MDA-MB-231 cells with low-HER2 level and the KPL4 cells with high-HER2 level. Two trastuzumab-insensitive breast cancer cell lines MDA-MB-231 (A) and KPL4 (B) were treated with trastuzumab (Tras) at different concentrations (0, 5, 10 µg/ml) for 48 hrs. Total RNA was prepared and analyzed by QRT-PC...

Heatmap of the differentially expressed miRNAs in response to trastuzumab treatment in HER2-overexpressing SKBr3 cells (A) and BT474 cells (B). To visualize the clusters of expressed miRNAs after trastuzumab treatment, hierarchical clustering was performed based on the differentially expressed miRNAs. Names of samples (top) and miRNAs (side) is sho...

Introduction: microRNAs (miRNAs), small noncoding RNAs, are deregulated in several diseases including cancer. miRNAs regulate gene expression at a posttranscriptional level by binding to 5'UTR, coding regions or 3'UTR of messenger RNAs (mRNA), inhibiting mRNA translation or causing mRNA degradation. The same miRNA can have multiple mRNA targets, a...

Epidermal growth factor (EGF) and its receptor play critical roles in non-small cell lung cancer (NSCLC) carcinogenesis. A functional polymorphism in the EGF gene has been linked to increased cancer susceptibility. This study aimed to evaluate the role of the EGF +61A/G polymorphism as risk factors in NSCLC patients. For the present case-control st...

Tiling array and novel sequencing technologies have made available the transcription profile of the entire human genome. However, the extent of transcription and the function of genetic elements that occur outside of protein-coding genes, particularly those involved in disease, are still a matter of debate. In this review, we focus on long non-codi...

MicroRNAs (miRNAs) can promote or inhibit tumor growth and are therefore being developed as targets for cancer therapies. They are diverse not only in the messenger RNAs (mRNA) they target, but in their production; the same hairpin RNA structure can generate mature products from each strand, termed 5p and 3p, that can bind different mRNAs. We analy...

Trastuzumab, a humanized monoclonal antibody directed against the extracellular domain of the HER2 oncoprotein, can effectively target HER2-positive breast cancer through several mechanisms. Although the effects of trastuzumab on cancer cell proliferation, angiogenesis and apoptosis have been investigated in depth, the effect of trastuzumab on micr...

Unlabelled: MicroRNAs (miRs) recently emerged as prominent regulators of cancer processes. In the current study we aimed at elucidating regulatory pathways and mechanisms through which miR-494, one of the miR species found to be down-regulated in cholangiocarcinoma (CCA), participates in cancer homeostasis. miR-494 was identified as down-regulated...

IQGAP1 knockout mice develop gastric cancer, but the IQGAP1 protein is associated with some advanced-stage human cancers. IQGAP1 expression is regulated by a microRNA, miR-124, through a binding site at the 3'-untranslated region, where a single nucleotide polymorphism (SNP) exists in the core binding region. We asked whether IQGAP1 expression is a...

Since 1993, when the first small non-coding RNA was identified, our knowledge about microRNAs has grown exponentially. In this review, we focus on the main progress in this field and discuss the most important findings under a historical perspective. In addition, we examine microRNAs as markers of disease diagnosis and prognosis, and as new therape...

Comment on: Pelletier C, et al. Cell Cycle 2011; 10:90-1:

Cell death and differentiation is a monthly research journal focused on the exciting field of programmed cell death and apoptosis. It provides a single accessible source of information for both scientists and clinicians, keeping them up-to-date with advances in the field. It encompasses programmed cell death, cell death induced by toxic agents, dif...

Commentary to:Identification of miRNAs that are associated with tumor metastasis in NeuroblastomaJigang Guo, Qian Dong, Zhixiang Fang, Xi Chen, Hongting Lu, Kehui Wang, Yuan Yin, Xing Cai, Nan Zhao, Jiangning Chen, Ke Zen, Junfeng Zhang and Chen-Yu Zhang

Minimally differentiated acute myeloid leukemia is heterogeneous in karyotype and is defined by immature morphological and molecular characteristics. This originally French-American-British classification is still used in the new World Health Organization classification when other criteria are not met. Apart from RUNX1 mutation, no characteristic m...

Medulloblastoma is the most common malignant brain tumor in children. The presence of microsatellite instability (MSI) in brain tumors, particularly medulloblastomas, has not been properly addressed. The aim of the present study was to evaluate the role of MSI in medulloblastoma carcinogenesis. MSI status was determined in 36 patients using a penta...