43 research outputs found
In vitro and in vivo properties of distinct populations of amniotic fluid mesenchymal progenitor cells
Human mesenchymal progenitor cells (MPCs) are considered to be of great promise for use in tissue repair and regenerative medicine. MPCs represent multipotent adherent cells, able to give rise to multiple mesenchymal lineages such as osteoblasts, adipocytes or chondrocytes. Recently, we identified and characterized human second trimester amniotic fluid (AF) as a novel source of MPCs. Herein, we found that early colonies of AF-MPCs consisted of two morphologically distinct adherent cell types, termed as spindle-shaped (SS) and round-shaped (RS). A detailed analysis of these two populations showed that SS-AF-MPCs expressed CD90 antigen in a higher level and exhibited a greater proliferation and differentiation potential. To characterize better the molecular identity of these two populations, we have generated a comparative proteomic map of SS-AF-MPCs and RS-AF-MPCs, identifying 25 differentially expressed proteins and 10 proteins uniquely expressed in RS-AF-MPCs. Furthermore, SS-AF-MPCs exhibited significantly higher migration ability on extracellular matrices, such as fibronectin and laminin in vitro, compared to RS-AF-MPCs and thus we further evaluated SS-AF-MPCs for potential use as therapeutic tools in vivo. Therefore, we tested whether GFP-lentiviral transduced SS-AF-MPCs retained their stem cell identity, proliferation and differentiation potential. GFP-SS-AF-MPCs were then successfully delivered into immunosuppressed mice, distributed in different tissues and survived longterm in vivo. In summary, these results demonstrated that AF-MPCs consisted of at least two different MPC populations. In addition, SS-AF-MPCs, isolated based on their colony morphology and CD90 expression, represented the only MPC population that can be expanded easily in culture and used as an efficient tool for future in vivo therapeutic applications
Origin and spread of human mitochondrial DNA haplogroup U7
Human mitochondrial DNA haplogroup U is among the initial maternal founders in Southwest Asia and Europe and one that best indicates matrilineal genetic continuity between late Pleistocene hunter-gatherer groups and present-day populations of Europe. While most haplogroup U subclades are older than 30 thousand years, the comparatively recent coalescence time of the extant variation of haplogroup U7 (~16–19 thousand years ago) suggests that its current distribution is the consequence of more recent dispersal events, despite its wide geographical range across Europe, the Near East and South Asia. Here we report 267 new U7 mitogenomes that – analysed alongside 100 published ones – enable us to discern at least two distinct temporal phases of dispersal, both of which most likely emanated from the Near East. The earlier one began prior to the Holocene (~11.5 thousand years ago) towards South Asia, while the later dispersal took place more recently towards Mediterranean Europe during the Neolithic (~8 thousand years ago). These findings imply that the carriers of haplogroup U7 spread to South Asia and Europe before the suggested Bronze Age expansion of Indo-European languages from the Pontic-Caspian Steppe region
Metabolic rewiring is associated with HPV-specific profiles in cervical cancer cell lines
Both HPV-positive and HPV-negative cervical cancers are associated with
aberrant metabolism, although the oncogenic drivers remain elusive. Here
we show the assessment of the metabolomic profiles of four distinct
cervical cell lines, a normal and three cancer cell lines, one
HPV-negative (C33A) and two HPV-positive (SiHa HPV16+, HeLa HPV18+),
employing an ultra performance liquid chromatography and a high
resolution mass spectrometry. Out of the total 462 metabolites, 248 to
326 exhibited statistically significant differences, while Random
Forests analysis identified unique molecules for each cell line. The two
HPV+ cell lines exhibited features of Warburg metabolism, consistent
with the role of the HPV E6 protein. SiHa and HeLa cells displayed
purine salvage pathway activity, while C33A cells revealed synthesis of
cytidine, via a novel mechanism. These data document a highly dynamic
HPV-specific rewiring of metabolic pathways occurring in cervical
cancer. Therefore, this approach can eventually provide novel
mechanistic insights into cervical carcinogenesis
Gene Therapy for Malignant and Benign Gynaecological Disorders: A Systematic Review of an Emerging Success Story
Despite the major advances in screening and therapeutic approaches, gynaecological malignancies still present as a leading cause of death among women of reproductive age. Cervical cancer, although largely preventable through vaccination and regular screening, remains the fourth most common and most lethal cancer type in women, while the available treatment schemes still pose a fertility threat. Ovarian cancer is associated with high morbidity rates, primarily due to lack of symptoms and high relapse rates following treatment, whereas endometrial cancer, although usually curable by surgery, it still represents a therapeutic problem. On the other hand, benign abnormalities, such as fibroids, endometriosis, placental, and embryo implantation disorders, although not life-threatening, significantly affect women’s life and fertility and have high socio-economic impacts. In the last decade, targeted gene therapy approaches toward both malignant and benign gynaecological abnormalities have led to promising results, setting the ground for successful clinical trials. The above therapeutic strategies employ both viral and non-viral systems for mutation compensation, suicide gene therapy, oncolytic virotherapy, antiangiogenesis and immunopotentiation. This review discusses all the major advances in gene therapy of gynaecological disorders and highlights the novel and potentially therapeutic perspectives associated with such an approach
Transcription factor ATF-3 regulates allele variation phenotypes of the human SLC11A1 gene
Genetic polymorphisms in the human solute carrier family 11 member 1
(SLC11A1) gene predispose to susceptibility to infectious/inflammatory
diseases and cancer. Human susceptibility to these diseases exhibits
allelic association with a polymorphic regulatory Z-DNA-forming
microsatellite of a (GT/AC)n repeat. The carriage of different alleles
may influence chromatin remodeling and accessibility by transcription
factors. Of particular importance is the binding site for the Activating
Protein 1 (AP-1) elements, (ATF-3 and c-Jun), adjacent to the 5’
sequence of the Z-DNA-forming polymorphism. The aim of the study was to
characterize the transcriptional mechanisms controlling different
alleles of SLC11A1 expression by ATF-3 and c-Jun. Allele 2,
[T(GT)(5)AC(GT)(5)AC(GT)(10)GGCAGA(G)(6)], and Allele 3,
[T(GT)(5)AC(GT)(5)AC(GT)(9)GGCAGA(G)(6)], were subcloned into the
PGL2Basic vector. Transient transfections of THP-1 cells with the
constructs, in the presence or absence of pATF-3 were preformed.
Luciferase expression was determined. To document the recruitment of
ATF-3 and c-Jun, to the polymorphic promoter alleles in vivo, we
performed ChIP assays with transient transfected THP-1 cells treated
with or without lipopolyssacharides. Our data documented that ATF-3
suppresses the transcriptional activation of Allele-3, and this
suppression is enhanced in the presence of lipopolyssacharides. Our
findings suggest that ATF-3 and c-Jun may influence heritable variation
in SLC11A1-dependent innate resistance to infection and inflammation
both within and between populations
The Q192R polymorphism of the paraoxonase-1 (PON1) gene is associated with susceptibility to gestational diabetes mellitus in the Greek population
A key factor protecting from oxidative stress in gestational diabetes
mellitus (GDM) and in type 2 diabetes (T2D) is paraoxonase-1 (PON1).
Inconclusive and limited data exist regarding the effect of a coding
polymorphism (Q192R) of the PON1 gene in conferring susceptibility to
both states. In the present study, we investigated the association
between the PON1 gene and the risk for GDM in the Greek population and
assessed for the first time its transcriptional efficiency. We studied
185 women with GDM and 104 non-diabetic controls for the PON1
polymorphism. For PON1 mRNA expression, peripheral leucocytes were
harvested from 20 GDM and 20 control women, harboring different
genotypes for the polymorphism, using real-time quantitative PCR. The RR
genotype and the R allele of the PON1 Q192R polymorphism were
significantly associated with an increased risk for GDM (p=0.012 and
p<0.0001, respectively). Furthermore, there was no statistical
correlation between the individual metabolic parameters tested and the
three genotypes. Finally, the expression levels of PON1 mRNA in GDM
patients did not exhibit any statistical difference compared with normal
controls (p=0.138). These data independently document that the Q192R
polymorphism is closely associated with GDM susceptibility, while the
PON1 gene expression is not impaired in GDM
The major circadian pacemaker ARNT-like protein-1 (BMAL1) is associated with susceptibility to gestational diabetes mellitus
Aims: Recently a relationship between circadian clock function and the
risk for type 2 diabetes (T2D) has been shown. BMAL1 is a key component
of the mammalian molecular clock. Two SNPs in the BMAL1 gene have been
identified to confer T2D susceptibility. In the present study we
investigated for the first time the association between the BMAL1 gene
and the risk for GDM, in a Greek population.
Methods: We studied 185 women with GDM and 161 non-diabetic controls for
BMAL1 polymorphisms. For BMAL1 mRNA expression, peripheral leukocytes
were harvested from 20 GDM and 20 control women, harboring different
genotypes for the tested polymorphisms, using real-time quantitative
PCR.
Results: The minor allele (A) of the BMAL1 rs7950226 (G > A)
polymorphism was found to be significantly associated with an increased
risk of GDM (P = 0.025). Analysis of the second BMAL1 rs11022775 (T > C)
polymorphism, showed that the C-allele frequency was strongly increased
in women with GDM (P = 4.455e-06). The CC genotype was also
significantly overrepresented in GDM vs. controls (P = 0.00001).
Additionally, the rs7950226G/rs11022775C and rs7950226A/rs11022775C
haplotypes were also found to be associated with increased
susceptibility to GDM. Furthermore, the expression levels of BMAL1 mRNA
were significantly lower in GDM patients than in controls.
Conclusion: These data suggest that the impairment of the BMAL1 clock
gene expression is closely associated with GDM susceptibility. (C) 2012
Elsevier Ireland Ltd. All rights reserved
Circadian clock gene expression is impaired in gestational diabetes mellitus
Dysfunction of the circadian clock genes is involved in the development
of obesity and type 2 diabetes (T2D). Since gestational diabetes
mellitus (GDM) and T2D share common genetic and phenotypic features, in
the present study, we investigated the status of the circadian clock in
a cohort of 40 Greek pregnant women with GDM, four with T2D and 20
normal controls. Peripheral blood mRNA transcript levels of 10 clock
genes (CLOCK1, BMAL1, PERI, PER2, PER3, PPARA, PPARD, PPARG, CRY1 and
CRY2) were determined by real-time quantitative PCR. GDM patients
expressed significantly lower transcript levels of BMAL1, PER3, PPARD
and CRY2 compared to control women (p < 0.05). No significant difference
was documented between GDM women maintained either under insulin
treatment or diet. A positive correlation was found between the
expression of BMAL1 versus CRY2 (r = 0.45, p = 0.003) and BMAL1 versus
PPARD (r = 0.43, p = 0.004). Further investigation on the functional
relevance of these clock genes, disclosed that expression of PER3
correlated negatively with HbA(1c) levels (r = -0.36, p = 0.022). These
data document for the first time that the expression of BMAL1, PER3,
PPARD and CRY2 genes is altered in GDM compared to normal pregnant women
and support the notion that deranged expression of clock genes may play
a pathogenic role in GDM